Effects of indoor air movement and ambient temperature on mosquito (Anopheles gambiae) behaviour around bed nets: implications for malaria prevention initiatives.

BACKGROUND: Until recently, relatively little research has been done on how mosquitoes behave around the occupied bed net in the indoor environment. This has been partly remedied in the last few years through laboratory and field studies, most of these using video methods and mosquito flight tracking. Despite these recent advances, understanding of the mosquito-bed net environment system, and the principles that underlie mosquito behaviour within it, is limited. This project aimed to further understand this system by studying the effects of gently moving air (such as might be introduced through room design to make the indoor environment more comfortable and conducive to ITN use) and warmer vs. cooler ambient conditions on mosquito activity around ITNs and other bed nets. METHODS: The activity of colonized female Anopheles gambiae around an occupied untreated bed net set up in a mosquito-proof tent in a large laboratory space was recorded under different ambient conditions using a laser detection-video recording system. Conditions tested were 'cool' (23-25 °C) and 'warm' (27-30 °C) air temperatures and the presence or absence of a cross-flow produced by a small central processing unit (CPU) fan pointed at the side of the net so that it produced a 'low-' or 'high-' speed cross-draught (approx. 0.1 and 0.4 m/s, respectively). Near-net activity in recordings was measured using video image analysis. RESULTS: In cool, still air conditions, more than 80% of near-net activity by An. gambiae occurred on the net roof. Introduction of the low-speed or high-speed cross-draught resulted in an almost total drop off in roof activity within 1 to 2 min and, in the case of the high-speed cross-draught, a complementary increase in activity on the net side. In warm, still conditions, near-net activity appeared to be lower overall than in cool, still air conditions and to be relatively less focussed on the roof. Introduction of the high-speed cross-draught in warm conditions resulted in a decrease in roof activity and increase in side activity though neither effect was statistically significant. CONCLUSIONS: Results are interpreted in terms of the flow of the stimulatory odour plume produced by the net occupant which, consistent with established principles of fluid dynamics, appears to rise quickly and remain more intact above the net occupant in cool, still air than in warm, still air. Cross-draught effects are ascribed to the changes they cause in the flow of the host odour plume as opposed to mosquito flight directly. The implications of these results for house designs that promote indoor air movement, on bed net design, and on other vector control measures are discussed. How mosquitoes approach a net is influenced both by indoor temperature and ventilation and their interaction. This system is in need of further study.

Mosquito Larval Abundance in Connected and Isolated Pools Beside a Stormwater Management Pond.

Twelve experimental pools (30 cm width × 30 cm depth) around a large stormwater management pond (SWMP) were used to test the hypothesis that small puddles of water similar to animal hoofprints or other irregularities support more abundant and diverse mosquito populations due to having fewer insect predators. Six of the 12 pools were connected to the SWMP by a deep channel (7 cm wide × 10 cm depth × 50 cm length). Mosquito larvae and potential predators were sampled weekly over 16 wk in the summer. More mosquito larvae were found in the isolated pools than in connected pools or in the pond itself (U = 5.5, z = 2.002, P = 0.045). The observed differences between isolated and connected pools are presented and results discussed in terms of SWMP design.

The effect of holes in long-lasting insecticidal nets on malaria in Malawi: results from a case-control study.

BACKGROUND: Long-lasting insecticidal nets (LLINs) are a cornerstone of malaria prevention. Holes develop in LLINs over time and compromise their physical integrity, but how holes affect malaria transmission risk is not well known. METHODS: After a nationwide mass LLIN distribution in July 2012, a study was conducted to assess the relationship between LLIN damage and malaria. From March to September 2013, febrile children ages 6-59 months who consistently slept under LLINs (every night for 2 weeks before illness onset) were enrolled in a case-control study at Machinga District Hospital outpatient department. Cases were positive for Plasmodium falciparum asexual parasites by microscopy while controls were negative. Digital photographs of participants' LLINs were analysed using an image-processing programme to measure holes. Total hole area was classified by quartiles and according to the World Health Organization's proportionate hole index (pHI) cut-offs [< 79 cm2 (good), 80-789 cm2 (damaged), and > 790 cm2 (too torn)]. Number of holes by location and size, and total hole area, were compared between case and control LLINs using non-parametric analyses and logistic regression. RESULTS: Of 248 LLINs analysed, 97 (39%) were from cases. Overall, 86% of LLINs had at least one hole. The median number of holes of any size was 9 [interquartile range (IQR) 3, 22], and most holes were located in the lower halves of the nets [median 7 (IQR 2, 16)]. There were no differences in number or location of holes between LLINs used by cases and controls. The median total hole area was 10 cm2 (IQR 2, 125) for control LLINs and 8 cm2 (IQR 2, 47) for case LLINs (p = 0.10). Based on pHI, 109 (72%) control LLINs and 83 (86%) case LLINs were in "good" condition. Multivariable modeling showed no association between total hole area and malaria, controlling for child age, caregiver education, and iron versus thatched roof houses. CONCLUSIONS: LLIN holes were not associated with increased odds of malaria in this study. However, most of the LLINs were in relatively good condition 1 year after distribution. Future studies should examine associations between LLIN holes and malaria risk with more damaged nets.

Behavioural responses of females of two anopheline mosquito species to human-occupied, insecticide-treated and untreated bed nets.

BACKGROUND: Insecticide-treated bed nets (ITNs), used extensively to reduce human exposure to malaria, work through physical and chemical means to block or deter host-seeking mosquitoes. Despite the importance of ITNs, very little is known about how host-seeking mosquitoes behave around occupied bed nets. As a result, evidence-based evaluations of the effects of physical damage on bed net effectiveness are not possible and there is a dearth of knowledge on which to base ITN design. METHODS: The dispersion of colony-raised female Anopheles gambiae and Anopheles albimanus was observed in 2-hr laboratory experiments in which up to 200 mosquitoes were released inside a mosquito-proof 3 m × 3 m tent housing a bed net arrayed with 18 30 cm × 30 cm sticky screen squares on the sides, ends and roof. Numbers of mosquitoes caught on the sticky squares were interpreted as the 'mosquito pressure' on that part of the net. RESULTS: Presence of a human subject in the bed net significantly increased total mosquito pressure on the net for both species and significantly re-oriented An. gambiae to the roof of the net. Anopheles albimanus pressure was greatest on the bed net roof in both host-present and no-host conditions. The effects of different human subjects in the bed net, of different ambient conditions (dry, cool conditions vs warm, humid conditions) and of bed net treatment (deltamethrin-treated or no insecticide) on mosquito pressure patterns were tested for both species. Species-specific pressure patterns did not vary greatly as a result of any of these factors though some differences were noted that may be due the size of the different human subjects. CONCLUSIONS: As a result of the interaction between host-seeking responses and the convective plume from the net occupant, species-specific mosquito pressure patterns manifest more or less predictably on the bed net. This has implications for bed net design and suggests that current methods of assessing damaged bed nets, which do not take damage location into account, should be modified.

Anatomy and function of the mouthparts of the biting midge, Culicoides sanguisuga (Diptera: Ceratopogonidae).

The mouthparts and upper food canal of the biting midge, Culicoides sanguisuga, are described with light, scanning electron, and transmission electron microscopy. The stylets (labrum, hypopharynx, mandibles, laciniae) are individually described so that their union into the piercingsucking syntrophium can be understoo. Labial anatomy and relationships to the syntrophium are also described. Syntrophial integrity is based on a complex system in which (1) the mandibles articulate with each other and with the hypopharynx; (2) the edges of the labrum are hooked over the sides of the mandibles; (3) the laciniae "clasp" the hypopharyngeal-mandibular complex to the labrum; and (4) the labellum holds the stylets together terminally. We propose that all muscles attached to the mandibles of the biting midge serve as retractors. This, combined with the fact that the mandibles have only one toothed edge (rearward-directed teeth on the lateral edge), suggests that retraction is the only active cutting stroke during biting. The mechanism contrasts with that of black flies (simuliids), which have mandibular protractors and retractors and teeth on both lateral and medial mandibular margins.

Fine structure of tarsal sensilla of male and female Simulium vittatum (Diptera: Simuliidae).

Scanning and transmission electron microscopic examination of the general and sensillar anatomy of the prothoracic tarsus of male and female Simulium vittatum reveals four kinds of hair-type sensilla: (1) generally distributed, mechanosensitive type 1 sensilla trichodea; (2) type 2 sensilla trichodea similar to type 1 though smaller and restricted to the ventral surfaces of tarsomeres 1-4; (3) triply-innervated, taste-sensitive peg sensilla of the ventral surfaces of tarsomeres 1-4; and (4) mid-laterally and terminally distributed type 1 sensilla chaetica that possess one mechanosensitive and four chemosensitive neurons. In addition, wholly internal chordotonal sensilla associated with the tarsal tendon are described. Regions of presumed was-secreting epithelial cells were found to line much of the inner ventral tarsal surface. Anatomical findings are discussed in terms of behavioral and/or physiological significance.

Mechanics of blood-feeding in black flies (Diptera, simuliidae).

The structure and interrelationships of the mouthparts and of the food canal and its accessory cephalic structures of the females of Simulium venustum are described through microscopic observations. The mouthparts that enter the would during feeding are the mandibles, maxillary laciniae, hypopharynx, and labrum and collectively form a "syntrophium." The labium and labellar lobes, which do not enter the wound, ensheathe the syntrophium distally and must be retracted to allow biting. We present an interpretation of mouthpart function during biting that emphasizes how biting steps are accomplished and what sensory structures are used to monitor the process. Four phases of biting are identified: (1) initial penetration of the skin effected by the mandibles; (2) consolidation of mouthpart position involving anchoring the syntrophium into the wound by means of the barbed laciniae; (3) diet sampling and active feeding-food (blood) is pumped by three groups of muscles forming two functional pumps, one located in the cibarium, the other in the pharynx. These pumps are separated from each other and from surrounding regions of the food canal by valve muscles making the pumping process a complex and highly coordinated series of muscular contractions; and (4) mouthpart disengagement involving removal of the laciniae, thus releasing the syntrophium from the wound.

Innervation and structure of mouth part sensilla in females of the black fly Simulium venustum (Diptera: Simuliidae).

Associated with the mouth parts of female Simulium venustum are 13 morphologic types of sensilla: four on the labium, seven on the labrum, one in the cibarium, and one on the genal process to which the mandibles articulate. Seven types are probably sensitive only to mechanicla cues and three only to chemical ones, whereas the other three probably function in both modes. These sensilla likely monitor feeding-associated chemical features of blood, sugar, and water and mechanical cues generated by the physical acts of ingestion. Each S. venustum female has approximately 450 chemosensitive and 230 mechanosensitive neurons in the mouthpart-associated sensilla. Both the total number of chemosensory neurons and the ratio of chemosensory to mechanosensory neurons in S. venustum are intermediate between those for blow flies, which feed on a wide variety of foodstuffs, and tsetse flies, exclusive blood-feeders. These differences may be related to whether determination of acceptability of a potential food source occurs at the site of feeding and is dependent upon simultaneous sensitivity to many chemical cues, as in blow flies, or is the result of a complex stimulus chain composed of all host-location steps and culminating with the detection of but a few phagostimulants in the food itself, as in blood-feeders.