Risk factors for malaria infection prevalence and household vector density between mass distribution campaigns of long-lasting insecticidal nets in North-western Tanzania.

BACKGROUND: Long-lasting insecticidal nets (LLINs) are the most widely deployed vector control intervention in sub-Saharan Africa to prevent malaria. Recent reports indicate selection of pyrethroid insecticide resistance is widespread in mosquito vectors. This paper explores risk factors associated with malaria infection prevalence and vector density between mass distribution campaigns, changes in net coverage, and loss of protection in an area of high pyrethroid resistance in Northwest Tanzania. METHODS: A cross sectional malaria survey of 3456 children was undertaken in 2014 in Muleba district, Kagera region west of Lake Victoria. Vector density was assessed using indoor light traps and outdoor tent traps. Anophelines were identified to species using PCR and tested for Plasmodium falciparum circumsporozoite protein. Logistic regression was used to identify household and environmental factors associated with malaria infection and regression binomial negative for vector density. RESULTS: LLIN use was 27.7%. Only 16.9% of households had sufficient nets to cover all sleeping places. Malaria infection was independently associated with access to LLINs (OR: 0.57; 95% CI 0.34-0.98). LLINs less than 2 years old were slightly more protective than older LLINs (53 vs 65% prevalence of infection); however, there was no evidence that LLINs in good condition (hole index < 65) were more protective than LLINs, which were more holed. Other risk factors for malaria infection were age, group, altitude and house construction quality. Independent risk factors for vector density were consistent with malaria outcomes and included altitude, wind, livestock, house quality, open eaves and LLIN usage. Indoor collections comprised 4.6% Anopheles funestus and 95.4% Anopheles gambiae of which 4.5% were Anopheles arabiensis and 93.5% were Anopheles gambiae sensu stricto. CONCLUSION: Three years after the mass distribution campaign and despite top-ups, LLIN usage had declined considerably. While children living in households with access to LLINs were at lower risk of malaria, infection prevalence remained high even among users of LLINs in good condition. While effort should be made to maintain high coverage between campaigns, distribution of standard pyrethroid-only LLINs appears insufficient to prevent malaria transmission in this area of intense pyrethroid resistance.

Studies on the resting behaviour and host choice of Anopheles gambiae and An. arabiensis from Muleba, Tanzania.

The relative efficacy of a mechanical (Prokopack) collection method vs. manual aspiration in the collection of resting mosquitoes was evaluated in northern Tanzania before and after an intervention using indoor residual spraying and longlasting insecticide-treated nets. In smoke-free houses mosquitoes were collected from the roof and walls, but in smoky houses mosquitoes were found predominantly on the walls. Anopheles gambiae (Diptera: Culicidae) constituted 97.7% of the 312 An. gambiae complex specimens identified before but only 19.3% of the 183 identified after the intervention. A single sampling with the Prokopack collected a third of the available insects. Anopheles gambiae completed its gonotrophic development indoors, whereas Anopheles arabiensis did so outdoors. In both species gonotrophic development took 2 days. Most unfed resting An. arabiensis collected outdoors were virgins, whereas the majority of engorged insects were parous (with well-contracted sacs). Daily survival was estimated to be 80.0%. Only 9.4% of the engorged An. arabiensis collected outdoors and 47.1% of those collected indoors had fed on humans. Using the Prokopack sampler is more efficient than manual methods for the collection of resting mosquitoes. Malaria transmission may have been affected by a change in vector composition resulting from a change in feeding, rather than reduced survival. Monitoring the proportions of members of the An. gambiae complex may provide signals of an impending breakdown in control.

Guidance for Evaluating the Safety of Experimental Releases of Mosquitoes, Emphasizing Mark-Release-Recapture Techniques.

Experimental releases of mosquitoes are performed to understand characteristics of populations related to the biology, ability to transmit pathogens, and ultimately their control. In this article, we discuss considerations related to the safety of experimental releases of living mosquitoes, applying principles of good practice in vector biology that protect human health and comfort. We describe specific factors of experimental releases of mosquitoes that we believe are critical to inform institutional biosafety committees and similar review boards to which proposals to conduct mosquito release experiments have been submitted. In this study, "experimental releases" means those that do not significantly increase vector capacity or nuisance biting relative to the unperturbed natural baseline. This document specifically does not address releases of mosquitoes for ongoing control programs or trials of new control methods for which broader assessments of risk are required. It also does not address releases of transgenic or exotic (non-native) mosquito species, both of which require particular regulatory approval. Experimental releases may include females and males and evaluation must consider their effects based on the number released, their genotype and phenotype, the environment into which they are released, and postrelease collection activities. We consider whether increases of disease transmission and nuisance biting might result from proposed experimental releases against the backdrop of natural population size variation. We recommend that experimental releases be conducted in a manner that can be reasonably argued to have insignificant negative effects. Reviewers of proposals for experimental releases should expect applicants to provide such an argument based on evidence from similar studies and their planned activities. This document provides guidance for creating and evaluating such proposals.

'Repel all biters': an enhanced collection of endophilic Anopheles gambiae and Anopheles arabiensis in CDC light-traps, from the Kagera Region of Tanzania, in the presence of a combination mosquito net impregnated with piperonyl butoxide and permethrin.

BACKGROUND: Mosquito nets containing synergists designed to overcome metabolic resistance mechanisms in vectors have been developed. These may enhance excitability in the mosquitoes and affect how they respond to CDC light-traps. Investigating the behaviour of vectors of disease in relation to novel mosquito nets is, therefore, essential for the design of sampling and surveillance systems. METHODS: In an initial experiment in Muleba, Tanzania, nine bedrooms from three housing clusters were sampled. CDC light-traps were operated indoors next to occupied untreated nets (UTN), Olyset® long lasting insecticidal net (LLIN) and Olyset Plus® LLIN containing piperonyl butoxide (PBO) synergist. Nets were rotated daily between the nine rooms over nine nights. A further series of experiments using the nets on alternate nights in a single room was undertaken during the short rains. Anopheles gambiae s.l. were collected in CDC light-traps, a window-trap and Furvela tent-trap. Anopheles gambiae s.l. were identified to species by polymerase chain reaction (PCR). RESULTS: In the initial experiment 97.7% of the 310 An. gambiae s.l. were An. gambiae s.s., the remainder being Anopheles arabiensis. The number of mosquitoes collected from 81 light-trap collections was greater in the presence of an Olyset [density rate ratio 1.81, 95% CI (1.22-2.67), p = 0.003] relative to an UTN. In a second experiment, in the wet season 84% of the 180 An. gambiae s.l. identified were An. arabiensis. The number of An. gambiae s.l. collected from a light-trap compared to a tent-trap was significantly higher when an Olyset Plus net was used compared to an UTN. Survival of the mosquitoes in the window trap was not reduced by the use of an Olyset Plus net in the bedroom relative to an Olyset net. CONCLUSION: Mosquitoes entering bedrooms, even those susceptible to pyrethroids, were not killed by contact with an Olyset Plus LLIN. The enhanced numbers of An. gambiae or An. arabiensis collected in light-traps when a treated net is used requires further experimentation and may be because of a heightened escape reaction on the part of the mosquito.

'Nature or nurture': survival rate, oviposition interval, and possible gonotrophic discordance among South East Asian anophelines.

BACKGROUND: Mosquito survival, oviposition interval and gonotrophic concordance are important determinants of vectorial capacity. These may vary between species or within a single species depending on the environment. They may be estimated by examination of the ovaries of host-seeking mosquitoes. METHODS: Landing collections, Furvela tent-trap and CDC light-trap collections were undertaken sequentially in four locations in Cambodia between February 2012 and December 2013 and samples from the collected mosquitoes were dissected to determine parity, sac stage (indicative of time spent prior to returning to feed) and egg stage. RESULTS: A total of 27,876 Anopheles from 15 species or species groups were collected in the four locations and 2883 specimens were dissected. Both the density and predominant species collected varied according to location and trapping method. Five species were dissected in sufficient numbers to allow comparisons between locations. Estimated oviposition interval differed markedly between species but less within species among different locations. Anopheles aconitus had the shortest cycle, which was 3.17 days (95 % CI 3-3.64), and Anopheles barbirostris had the longest cycle, which took four days (95 % CI 3.29-4). Anopheles minimus had a higher sac rate in weeks leading up to a full moon but there was apparently little effect of moon phase on Anopheles dirus. Despite the fact that many of the species occurred at very low densities, there was no evidence of gonotrophic dissociation in any of them, even during sustained hot, dry periods. The principal Cambodian malaria vector, An. dirus, was only common in one location where it was collected in miniature light-traps inside houses. It did not appear to have an exceptional survival rate (as judged by the low average parous rate) or oviposition cycle. CONCLUSIONS: Differences in the oviposition interval were more pronounced among species within locations than within species among ecologically diverse locations. A nationwide survey using CDC light-traps for the collection of An. dirus inside houses may help in determining patterns of malaria transmission in Cambodia.

Effects of the spatial repellent metofluthrin on landing rates of outdoor biting anophelines in Cambodia, Southeast Asia.

The emergence of artemisinin-resistant malaria in Southeast Asia is a major problem. The fact that many people become infected with malaria when they are outside has prompted the development of 'spatial' rather than topical repellents. The respective effects of one or four slow-release emanators of metofluthrin, a pyrethroid, were tested in Pailin, Pursat and Koh Kong, Cambodia. Numbers of mosquitoes counted in outdoor landing catches when one or four emanators were suspended close to the collector were compared with control collections. In Pailin, the effects of emanators on catches in Furvela tent traps and Centers for Disease Control (CDC) light traps suspended underneath houses were also investigated. Rate ratios were used to determine differences. A total of 29 255 mosquitoes were collected over 2934 h of landing collections, 87 nights of tent trapping and 81 nights of light trap capture. In Pailin, landing rates were reduced by 48% by a single emanator and by 67% by four emanators (P < 0.001). Similar reductions were observed in the number of mosquitoes collected in tent traps and the number of anophelines only collected in light traps. Results were similar in Pursat, but, for unknown reasons, those in Koh Kong showed no difference between control and metofluthrin collections (P > 0.05). These findings suggest that although the product can produce a significant effect, it requires further improvement.

Analysis of the sporozoite ELISA for estimating infection rates in Mozambican anophelines.

Comparisons were undertaken to investigate cost-effective methods of implementing the enzyme-linked immunosorbent assay (ELISA) for sporozoite determination in anophelines when large numbers require processing. Comparisons between ELISA plate reader and visual assessments were performed with Anopheles funestus and Anopheles gambiae s.l. (Diptera: Culicidae), as were comparisons between whole-body mosquito samples, heads and thoraces, and abdomens alone. Rates obtained from pools of five or 10 mosquitoes were compared with those for individual mosquitoes, as were rates obtained using different sampling methods. A total of 41 792 An. funestus and 9431 An. gambiae s.l. collected in light traps, and 22 323 An. funestus and 6860 An. gambiae s.l. from exit collections were analysed. Visual assessments gave results similar to those of machine readings. Sporozoite rates were similar in both species, as were rates by collection method. The use of whole mosquitoes increased estimates of infection rate by 0.6%. Pool size did not affect infection rates of An. gambiae s.l., but rates were higher among individually tested An. funestus than among those tested in pools. For large-scale surveys, the use of whole mosquitoes in pools of 10 mosquitoes, with correction for overestimation, and the noting of results according to a simple three-stage visual assessment of positivity is the most cost-effective approach and is sufficient to obtain reliable data for comparative purposes.

Evidence of an 'invitation' effect in feeding sylvatic Stegomyia albopicta from Cambodia.

BACKGROUND: Orientation of haematophagous insects towards a potential host is largely mediated by kairomones that, in some groups or species may include chemicals produced during feeding by the insects themselves, the so called 'invitation' effect. METHODS: The 'invitation' effect in blood-feeding diurnally active Stegomyia albopicta was investigated over 33 days in secondary forest in Mondolkiri Province, Cambodia. Two human volunteers sitting inside a shelter collected mosquitoes and noted where and when they landed. A 10% emanator of a synthetic pyrethroid with high vapour action was in use on alternate days. RESULTS: Overall, 2726 mosquitoes were collected, 1654 of which had the landing site recorded. The heads of the volunteers were the locations with the highest density of landings per surface area whilst the knees and elbows accounted for most of the landings received on the arms and legs. Landings recorded within three minutes of each other on a collector were about 2.5 times more likely to be on the same body part than on a random body part, weighted for landing site preference. This preference did not vary with collector or pyrethroid. CONCLUSIONS: The 'invitation' effect may be due to a semio-chemical produced early in the feeding process. Incorporation of such a chemical into traps designed to control this important vector of dengue and chikungunya viruses might potentially improve their attractiveness.

Living on the edge: a longitudinal study of Anopheles funestus in an isolated area of Mozambique.

BACKGROUND: Understanding the survival strategies of malaria vectors at the edges of their distribution, where they are under stress from environmental conditions, may lead to the development of novel control techniques and may help predict the effects of climate change on these mosquitoes. METHODS: The population dynamics of an isolated population of Anopheles funestus from the peninsula of Linga Linga in southern Mozambique was studied over a period of 104 weeks from March 2009 to May 2011 by 917 light-trap and 390 exit collections, mostly in an area close to a seasonal pond. RESULTS: Over the sampling period, 3,684 An. funestus females were caught. Densities decreased with increasing distance from the pond. In 2009 and 2010, a single annual peak in An. funestus density coincident with the single annual peak in rainfall was observed, but a clear population peak was absent during the first 21 weeks of 2011. In between population peaks, An. funestus remained present at low densities. In light trap collections, the proportion of gravid mosquitoes was significantly higher during the 'low season' (the period between peaks) than during the peak season (RR = 4.3, p<0.001). In contrast, in exit collections, the proportion of gravid mosquitoes was significantly lower during low season than during the peak season (RR = 0.64, p<0.01). Also, in light traps, the proportion of part-fed females was higher during the low season than during the peak season (RR = 4.5, p<0.001), whereas this was inversed for engorged females (RR = 0.46, p<0.05).Thirteen out of 289 (4.5%) An. funestus tested positive in the sporozoite ELISA. The proportion of sporozoite positive females was higher during the low season (6.25%, six out of 96) than during the peaks (3.63%, seven out of 193), but this difference was not significant. CONCLUSIONS: It is suggested that a proportion of the mosquito population may become gonotrophically discordant during the long dry season resulting in enhanced mosquito survival and sustained malaria transmission.

Population dynamics and spatial structure of human-biting mosquitoes, inside and outside of houses, in the Chockwe irrigation scheme, southern Mozambique.

Focal control of malaria vectors, a potentially cost-effective alternative to conventional control, requires a spatio-temporal understanding of the mosquitoes. Trapping of African malaria vectors has generally been limited to inside houses making distribution estimates dependent on the location of dwellings. The development of tent-traps to sample outdoor biting mosquitoes has enabled more independent estimates. Here we describe both temporal and spatial variation in mosquito movements in an irrigation project village in southern Mozambique. Six hundred and ninety-three tent-trap collections (525 of which were paired with light-trap collections), 552 exit collections and 391 collections of mosquitoes resting inside houses were undertaken from March 2005 to April 2006. Fifteen species of mosquito were collected (five exclusively as larvae). Mansonia africana was the most common finding, numbers being greatest away from the village. Only Anopheles funestus, An. tenebrosus and Culex quinquefasciatus were collected in greater numbers in light-traps compared to tent-traps. Among the common mosquitoes, correlations in numbers of mosquito collected in paired tent and in light-traps were significant for all but An. tenebrosus. Inverse distance weighting was used to produce raster density maps of the most common mosquitoes. All species, with minor variations, in both hot and cool seasons, were collected in greatest numbers close to the edges of the village where water suitable for larval development was available. All exophilic anophelines species tested negative for sporozoites. It is suggested that focal control of larvae, applied by the villagers themselves, could be a suitable alternative to conventional control in this and similar villages.

Netting barriers to prevent mosquito entry into houses in southern Mozambique: a pilot study.

BACKGROUND: One of the best ways to control the transmission of malaria is by breaking the vector-human link, either by reducing the effective population size of mosquitoes or avoiding infective bites. Reducing house entry rates in endophagic vectors by obstructing openings is one simple way of achieving this. Mosquito netting has previously been shown to have this effect. More recently different materials that could also be used have come onto the market. Therefore, a pilot study was conducted to investigate the protective effect of three types of material against Anopheles funestus and Anopheles gambiae s.l entry into village houses in Mozambique when applied over the large opening at the gables and both gables and eaves. METHODS: A two-step intervention was implemented in which the gable ends of houses (the largest opening) were covered with one of three materials (four year old mosquito bed nets; locally purchased untreated shade cloth or deltamethrin-impregnated shade cloth) followed by covering both gable ends and eaves with material. Four experimental rounds (each of three weeks duration), from four houses randomly assigned to be a control or to receive one of the three intervention materials, were undertaken from March to August 2010 in the village of Furvela in southern Mozambique. Mosquito entry rates were assessed by light-trap collection and the efficacy of the different materials was determined in terms of incidence rate ratio (IRR), obtained through a Generalized Estimating Equations (GEE), of mosquito entry in a treated house compared to the untreated (control) house. RESULTS: Altogether 9,692 An. funestus and 1,670 An. gambiae s.l. were collected. Houses treated with mosquito netting or the untreated shade cloth had 61.3% [IRR = 0.39 (0.32-0.46); P <0.0001] and 70% [IRR = 0.30 (0.25 - 0.37); P <0.001] fewer An. funestus in relation to untreated houses, but there was no difference in An. funestus in houses treated with the deltamethrin-impregnated shade cloth [IRR = 0.92 (0.76 -1.12); P = 0.4] compared to untreated houses. Houses treated with mosquito netting reduced entry rates of An. gambiae s.l, by 84% [IRR = 0.16 (0.10 - 0.25); P <0.001], whilst untreated shade cloth reduced entry rates by 69% [IRR = 0.31 (0.19 -0.53); P <0.001] and entry rates were reduced by 76% [IRR = 0.24 (0.15 0.38); P <0.001] in houses fitted with deltamethrin-impregnated shade cloth.

Some like it cool: the effect of ambient temperature on the size of Anopheles funestus from southern Mozambique.

The wing lengths of 3,553 unfed, recently emerged, and 13,256 gravid female Anopheles funestus Giles from exit collections, undertaken between March 2004 and May 2005, were measured. Only in the warmest months were mean wing lengths of unfed females significantly smaller than gravid females. Mean wing lengths of unfed, females varied from 2.26 mm (Bootstrap 95% CI [2.25-2.27]) in January when mean air temperatures were 24.8 degrees C to 2.70 mm (Bootstrap 95% CI [2.68-2.72]) in July when mean temperatures were 8.4 degrees C cooler. Mean wing lengths increased by 0.05 mm for each degree decrease in air temperature. Temperature explained 60% of the variation in wing length.

The effect of rainstorms on adult Anopheles funestus behavior and survival.

We describe the effect that the passage of a cold front, with a subsequent heavy rainstorm ten days later, had on a population of Anopheles funestus mosquitoes collected exiting houses or in light-traps from a village in southern Mozambique. Temperature effects explained 40% (r=0.634; p <0.001) of the variation in numbers of males collected and 19% of the variation in gravid females collected (r=0.437; p=0.033). The age structure of mosquitoes varied according to distance from the breeding site (χ(2) = 64.1, df 6, p <0.001). The proportion of parous insects that were caught in the light-traps with sacs (χ(2) = 6.33, d.f. 2, p=0.042) and young insects that had mated before being collected (χ(2) = 13,3, d.f. 2, p=0.001) were reduced on the night of the rain but this effect was short lived. It is concluded that the effect of rain on mosquito populations depends on the kind of water body used for larval development.

Artemisinin-based combination therapy does not measurably reduce human infectiousness to vectors in a setting of intense malaria transmission.

BACKGROUND: Artemisinin-based combination therapy (ACT) for treating malaria has activity against immature gametocytes. In theory, this property may complement the effect of terminating otherwise lengthy malaria infections and reducing the parasite reservoir in the human population that can infect vector mosquitoes. However, this has never been verified at a population level in a setting with intense transmission, where chronically infectious asymptomatic carriers are common and cured patients are rapidly and repeatedly re-infected. METHODS: From 2001 to 2004, malaria vector densities were monitored using light traps in three Tanzanian districts. Mosquitoes were dissected to determine parous and oocyst rates. Plasmodium falciparum sporozoite rates were determined by ELISA. Sulphadoxine-pyrimethamine (SP) monotherapy was used for treatment of uncomplicated malaria in the contiguous districts of Kilombero and Ulanga throughout this period. In Rufiji district, the standard drug was changed to artesunate co-administered with SP (AS + SP) in March 2003. The effects of this change in case management on malaria parasite infection in the vectors were analysed. RESULTS: Plasmodium falciparum entomological inoculation rates exceeded 300 infective bites per person per year at both sites over the whole period. The introduction of AS + SP in Rufiji was associated with increased oocyst prevalence (OR [95%CI] = 3.9 [2.9-5.3], p < 0.001), but had no consistent effect on sporozoite prevalence (OR [95%CI] = 0.9 [0.7-1.2], p = 0.5). The estimated infectiousness of the human population in Rufiji was very low prior to the change in drug policy. Emergence rates and parous rates of the vectors varied substantially throughout the study period, which affected estimates of infectiousness. The latter consequently cannot be explained by the change in drug policy. CONCLUSIONS: In high perennial transmission settings, only a small proportion of infections in humans are symptomatic or treated, so case management with ACT may have little impact on overall infectiousness of the human population. Variations in infection levels in vectors largely depend on the age distribution of the mosquito population. Benefits of ACT in suppressing transmission are more likely to be evident where transmission is already low or effective vector control is widely implemented.

Feeding frequency and survival of Anopheles gambiae in a rice-growing area in Ghana.

Mortality rates, determined by dissection, of predominantly M form female Anopheles gambiae (Diptera: Culicidae) were estimated. Mosquitoes were collected in tent traps and light traps in an irrigation project village in Ghana in June and July 2010, when much of the area was flooded. Both M and S form larvae were collected from rice fields (74 of 80 specimens were M form). Adults were collected in equal proportions from the two traps (90 of 107 specimens from the light trap and 106 of 116 specimens from the tent trap were M form). During the study, collection numbers rose from 105 to 972 per night. A total of 1787 of the 15 431 An. gambiae collected were dissected. Of these, 953 (53%) were found to have taken their first bloodmeal, either as virgins or following mating. The age profiles of mosquitoes collected alive and dead, respectively, were similar. Eighteen of 2933 (0.61 ± 0.49%) specimens were found to be positive for sporozoites in an enzyme-linked immunosorbent assay (ELISA). Lagged cross correlations among the different age groups implied that the mosquitoes fed on days 2 and 4 following emergence prior to oviposition and every 2.65 ± 0.17 days thereafter. The best model to describe the observed population patterns implied a daily mortality of 84%. The results are discussed in relation to possible mosquito control measures for the village.

A novel experimental hut for the study of entrance and exit behaviour of endophilic malaria vectors.

Experimental huts have played a long and illustrious role in the study of mosquito vectors of disease. Here we present a design of a novel, cheap, 12 sided, experimental hut based on the African roundhouse. The panels covering each side can open and act as a door and above each panel there is a shutter that can also open. This allows for a great combination of possible openings for the study of entry and exit behaviour of mosquitoes. Preliminary results describing the exit behaviour of Anopheles funestus when four of the panels were open are described.

Studies on the bionomics of male Anopheles gambiae Giles and male Anopheles funestus Giles from southern Mozambique.

Little is known about the fitness of wild male mosquitoes, the females of which are vectors of malaria. The problem of studying male biology has been exacerbated by difficulties associated with catching them. In southern Mozambique, however, almost the entire adult population of An. funestus and An. gambiae s.l. rest inside houses. They leave in a dusk exodus, which makes them easy to collect. In 8,348 exit collections from a village from 2003 to 2009, 567,195 male An. funestus and 34,591 male An. gambiae s.l. were collected. During the study, numbers of An. funestus increased but numbers of An. gambiae s.l. declined to the point of extinction. Overall numbers of An. gambiae s.l. were positively correlated with temperature, whilst the relationship between temperature and numbers of An. funestus changed from an initially positive one in the first three years of the study to a negative one in the last three years. Marked males were recaptured up to 300 m from the release site, with most recaptures occurring within 150 m. Estimated mean daily survival of male An. funestus was 0.86 (95% C.I. 0.869-0.850). For the years 2003-2007, estimated mean daily survival of male An gambiae s.l. was 0.660 (95% C.I. 0.682-0.638). For either species, there was no relationship between mean weekly temperature and estimated daily survival. These results imply that males of An. funestus live as long as females but have a relatively short flight range. They are discussed in the light of possible release strategies of sterile or genetically modified mosquitoes.

Do developing malaria parasites manipulate their mosquito host? Evidence from infected Anopheles funestus (Giles) from Mozambique.

Mosquito survival is linked to the activities performed in each oviposition cycle, whilst development of malaria parasites in them is largely temperature dependent. Extending the oviposition cycle of the mosquito, even as a side effect of normal development of the parasite, may enhance malaria transmission. A study was therefore undertaken to compare the time spent before returning to feed among infected and uninfected host seeking Anopheles funestus from a village in southern Mozambique. The stomachs of 2073 parous mosquitoes (1289 with sacs, indicative of a rapid return to host-seeking and 784 without sacs indicating a delay in host-seeking) from four extended sampling periods were examined for oocysts. From three of these, and overall, significantly more mosquitoes without sacs had oocysts than mosquitoes with sacs (Fishers Exact Test P=0.0051). This is the first time that possible effects on oviposition cycle length among infected vectors of human malaria have been demonstrated.

Studies on the behaviour of peridomestic and endophagic M form Anopheles gambiae from a rice growing area of Ghana.

The 'paddy paradox', the occurrence of large populations of vectors but low amounts of malaria transmission where irrigated rice is grown, was investigated in a village in Ghana where M form Anopheles gambiae are common. Peridomestic and indoor host-seeking mosquitoes were collected in tent traps and light traps over 21 consecutive nights at the start of the rainy season in June 2009 when the population increased exponentially from less than 100 per night to over 1000. Infection rates in the overall mosquito population were 0.3% and in the estimated parous population were 1.9%. Numbers of An. gambiae in the tent trap peaked between midnight and 02:40 am. The majority of insects were taking their first blood meal, as virgins or shortly after mating. More than expected were collected in the light trap during a rainstorm at the start of the rains but overall numbers were not affected. Fewer than expected were collected after a subsequent storm. Recruitment to the adult population decreased over the following days. It is hypothesised that the 'paddy paradox' is due to young pre-gravid insects dispersing more widely than gravid ones, not necessarily to low survival in the mosquito.

Does moonlight influence the biting behaviour of Anopheles funestus?

The possible effect of moonlight on the biting behaviour of mosquitoes in southern Mozambique, in particular that of Anopheles funestus (Diptera: Culicidae), a primary vector of malaria, was investigated by comparing catches indoors and outdoors using CDC light traps and 'Furvela' tent traps, respectively, for 35 consecutive nights, from 9 September to 15 October 2008. Collections were separated into three 4-hourly samples each night. A total of 17 591 mosquitoes belonging to nine species were collected, 6747 in light traps and 10 844 in tent traps. Anopheles funestus (n = 7634) and Mansonia africana (n = 4859) were the most abundant species. Fluctuations in temperature and humidity were the two environmental variables associated with changes in relative abundance of mosquitoes. Most An. funestus were collected indoors, with the majority collected in the first 4 h of the night. This was most evident on nights when moonlight was present in the early part of the night. A total of 3488 An. funestus were dissected for gonotrophic age determination. Parous rates did not change with lunar phase, but estimated oviposition cycle length was significantly shorter on nights when moonlight was present at the time of oviposition. Moonlight at dusk did not, however, affect the proportion of newly emerged insects with mating plugs collected. Outdoor transmission of malaria, especially on moonlit nights, remains a problem for control programmes.