Impact of untreated bednets on prevalence of Wuchereria bancrofti transmitted by Anopheles farauti in Papua New Guinea

Abstract Despite the growing evidence that insecticide‐treated mosquito nets reduce malaria morbidity and mortality in a variety of epidemiological conditions, their value against lymphatic filariasis infection and disease is yet to be established. The impact of untreated bednets on the prevalence of Wuchereria bancrofti (Cobbold) (Nematoda: Filarioidea) infection and disease was investigated on Bagabag island in Papua New Guinea, where both malaria and filariasis are transmitted by the same vector mosquitoes of the Anopheles punctulatus Dönitz group (Diptera: Culicidae). Community‐wide surveys were conducted recording demographic characteristics including bednet usage. Physical examinations for hydrocoele and lymphoedema were performed and blood samples assessed for filarial and malaria parasites. Mosquitoes were sampled using the all‐night landing catch method and individually dissected to determine W. bancrofti infection and infective rates. Bednet usage among residents was 61% and the mean age of users (25.6 years) was similar to non‐users (22.5 years). Anopheles farauti Laveran was the only species were found to contain filarial larvae: 2.7% infected (all stages), 0.5% infective (L3). The overall W. bancrofti microfilaraemia and antigenaemia rates were 28.5% and 53.1%, respectively. Bednet users had lower prevalence of W. bancrofti microfilaraemia, antigenaemia and hydrocoele rates than non‐users. In comparison, untreated bednets had no effect on the prevalence and intensity of Plasmodium falciparum and P. vivax infections. The impact of bednet usage on rates of microfilaraemia and antigenaemia remained significant even when confounding factors such as age, location and sex were taken into account, suggesting that untreated bednets protect against W. bancrofti infection.     

Permethrin-impregnated bednets are more effective than DDT house-spraying to control malaria in Solomon Islands

Abstract. A field trial compared DDT house-spraying with permethrin-impregnated bednets for malaria control in Solomon Islands from 1987 to 1991. Mortality-rates of malaria vector Anopheles farauti in exit window traps were 11.6% from an untreated hut, 10.1% from a hut sprayed with DDT 2 g/m², and 98% of those from a hut in which the occupants used bednets treated with permethrin 0.5 g/m². Since bioassays of the DDT-sprayed walls (15 min exposure in W.H.O. standard test cones) gave 77% mortality of Anfarauti , it was concluded that the insignificant impact of DDT could be explained by the exophilic behaviour of endophagic vectors, whereas the greater impact of permethrin was attributed to the more effective exposure of Anfarauti females to the impregnated bednets - attracted by the occupants. The parous rate was higher indoors, except in the area with permethrin-impregnated bednets. It was therefore concluded that permethrin-impregnated bednets reduced the mean longevity of Anfarauti and hence its vectorial capacity. The circumsporozoite (CS) antigen positivity rate of Anfarauti in the DDT area was 0.18% outdoors, significantly less than 1.42% indoors. In the comparison area CS rates were 0.65% outdoors and 0.75% indoors. CS antigen was not detected in Anfarauti from the bednet area, indicating the apparent prevention of malaria transmission. As DDT spraying was so much less effective, it was discontinued in 1993 and permethrin-impregnated bednets are now the principal malaria control method in Solomon Islands.     

Behaviour of Anopheles albimanus in relation to pyrethroid-treated bednets

Abstract. Responses of the malaria vector Anopheles albimanus to pyrethroid impregnated bednets made of cotton or nylon, compared with untreated nets, were investigated in houses occupied by two people inside and/or outside two bednets, in coastal Chiapas, México. The pyrethroid used was lambdacyhalothrin 30mga.i./m². Bioassay mortality rates of An.albimanus exposed to treated nets for 3 or 15min, rose from 40–55% to 90–100% for nylon nets 3–19 weeks post-treatment, but were consistently lower for treated cotton nets. An.albimanus females (collected unfed on human bait) were released in houses surrounded by curtains for trapping mosquitoes that exited from the house. Floor sheets were used in and around each experimental house for retrieving any mosquitoes knocked-down and/or killed. During post-treatment assessment for 17 weeks, An.albimanus blood-feeding success rates were 23–24% with untreated nets, 14–18% with treated cotton nets and 8–15% with treated nylon nets, significantly reduced when both human baits were inside the treated bednets, but not when one or more baits were outside the treated bednet(s) within the house. Proportions of mosquitoes leaving houses <3 h post-release were 53–59% from houses with untreated bednets versus 65–78% with treated bednets. Except in one case (when both humans were outside treated cotton nets), these increased early exit rates were significant, whether or not the human baits were inside the treated bednets indoors. Mortality rates of An.albimanus females exiting overnight (22.00–06.00 hours) averaged 15–39% from houses with treated cotton and 16–46% with treated nylon nets, very significantly greater than the control mean rates of 6–8% mortality with untreated nets. Observations on wild-caught An. albimanus females marked with fluorescent powder and released indoors revealed that few mosquitoes (3–11 %) actually contacted the bednets unless both human baits remained under them - when contact rates were 22% on treated nylon, 23% on treated cotton and 42% on untreated nets (P= 0.05). The mean resting time was significantly longer on untreated (14.4min) than on treated nylon (5.8min) or cotton (9.5 min) bednets, whereas mean resting times on other surfaces indoors were 16.5–19.8min. Proportions exiting within 2h of release were significantly more from houses with treated houses (33–35%) than with untreated nets (8%). However, mortality rates of mosquitoes that landed on treated nets were very significantly greater (90–100%) than after landing on untreated nets (10%). Thus, despite some excito-repellency, lambdacyhalothrin-impregnated bednets (especially made of nylon) proved to be effective as an alternative to house-spraying against An.albimanus.     

Seasonal abundance and biting behaviour of Anopheles punctulatus and An.koliensis in Malaita Province, Solomon Islands, and a trial of permethrin impregnated bednets against malaria transmission

Seasonal abundance of the malaria vectors Anopheles punctulatus Donitz and An.koliensis Owen in Bilimanu, an isolated inland village with forty-two houses in Malaita Province of the Solomon Islands, was monitored over 28 months by means of all-night landing/biting catches at one site during June 1985 to September 1987. Totals of 1250 An.punctulatus and 141 An.koliensis were collected, the latter being the largest number of this species ever caught at any locality in the Solomons. Bednets impregnated with permethrin 0.5 g/m² were introduced in December 1986 to be used at night by all 190 villagers for protection against malaria vectors. Bioassay tests with An.punctulatus blood-fed females exposed under nets for lOmin resulted in 100% mortality up to 50 weeks post-impregnation. For An.punctulatus, the main vector species, the mean catch (indoors + outdoors) per man hour was 2.9 (range 0.7–13.2) before a cyclone on 19 May 1986, and, 0.66 (0.2-2.7) after the cyclone. The vector survival rates were usually high before the cyclone, but erratically lower thereafter for An.punctulatus. An.koliensis disappeared after the cyclone. Both An.punctulatus and An.koliensis consistently showed higher rates of biting man indoors than outdoors and their diel biting cycle showed a peak around midnight. Outdoors, the parous proportion of An.punctulatus was twice the nulliparous, and nearly so indoors. Following intervention with permethrin-treated bednets, the mean catch of An.punctulatus fell to 0.35 per man-hour (monthly range 0–1.5). The Plasmodium falciparum malaria infection rate reduced from 10% pre-intervention to zero in September 1987, 9 months after intervention, and then rose again. With re-impregnation of bednets in November 1987, the P.falciparum infection rate was again reduced by March 1988. No effect of treated bednets was observed on the prevalence of P.vivax malaria.     

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.     

The campaign against malaria in central western Madagascar: comparison of the efficacy of lambda-cyhalothrin and DDT house spraying. I--Entomological study]

For malaria vector control in Madagascar, the efficacy of lambda-cyhalothrin 10% wettable powder (ICON 10 WP) was compared with DDT 75% WP for house-spraying. This evaluation was conducted from November 1997 to September 1998 in highland villages of Vakinankaratra Region, at the fringe of the malaria epidemic zone, outside the zone covered by routine DDT house-spraying (Opération de pulvérisation intro-domiciliaire de DDT: OPID zone). Treatments were compared by house-spraying in four areas: 1) application of DDT 2g ai/m2 and 2) lambda-cyhalothrin 30 mg ai/m2 in previously unsprayed villages; 3) no intervention (control); 4) OPID 5th cycle of DDT 2g ai/m2. The prevalent vector Anopheles funestus almost disappeared from both the DDT and ICON sprayed areas, whereas in the unsprayed (control) area An. funeslus density went up to 60 females per room in April and there were two seasonal peaks of malaria transmission in January and March (see following paper). In the area sprayed with ICON, the parous rate of An. funestus decreased from 47% pre-spray to 39% six months post-spraying, while the parous rate increased in DDT-sprayed area (from 57% pre-spray to 64% six months post-spray). Bioassays of An. funestus on treated walls, six months post-spray, gave mortality rates of 100% on DDT and 90% on ICON. Conversely, ICON appeared to be more effective than DDT on thatched roofs (66% versus 100%, respectively, six months post-spray). In areas sprayed with DDT or ICON the density of An. arabiensis were little affected. This study demonstrated that, under equivalent conditions, both DDT and lambda-cyhalothrin were effective in reducing malaria transmission on the western fringes of the malaria epidemic zone of the malagasy highlands, with a residual effect lasting at least for six months. Lambda-cyhalothrin appeared to be more effective than DDT in reducing the longevity of malaria vectors. In addition to efficacy, the choice of insecticide for malaria vector control should take into account their acceptability by human populations and their toxicity and persistence in the environment.     

The influence of permethrin-impregnated bednets and mass drug administration on the incidence of Plasmodium falciparum malaria in children in Sabah, Malaysia

A small-scale trial was carried out in the Upper Kinabatangan district of Sabah, Malaysia, to determine the effect of using permethrin-impregnated bednets on malaria transmission. A total of 306 nylon bednets with cotton borders, impregnated at a dose estimated to have been 0.062 g permethrin/m2 of nylon netting, were distributed to 139 households in five villages. At the time of distributing bednets, mass drug administration with Fansidar plus primaquine was also administered to the human population to clear all parasitaemias due to Plasmodium falciparum Welch. In another village, for comparison, mass drug administration was the only intervention. After intervention measures in December 1984 and January 1985, the parasite rates in children declined in all villages during the first month, significantly more in the villages with impregnated bednets than in the control, thus proving that the nets had an impact on malaria. However, after about 2 months, parasite rates started to increase again. After 4-6 months, parasite rates in the villages with bednets approached the rate in the control village without nets. The increase in parasite rates was paralleled by a significant deterioration in the quality, physical condition and the degree of non-utilization of bednets. Entomological evaluation proved the efficacy of permethrin-impregnated nets for controlling Anopheles balabacensis Baisas and other anophelines. Bioassays (1 h exposure) of permethrin-impregnated bednets gave 100% mortality initially and 44-61% mortality after 85-106 days. Mosquito collections in treated bednets were significantly reduced for at least 217 days. The project failed to achieve prolonged suppression of malaria transmission for a combination of entomological, sociological and practical reasons which are discussed in relation to the objectives and implementation of future bednet studies.     

Effects of community-wide use of lambdacyhalothrin-impregnated bednets on malaria vectors in rural Sierra Leone

Abstract. The effect of community-wide use of bednets treated with lambdacyhalothrin 10mg/m² on the malaria vector Anopheles gambiae (forest form) was evaluated in Sierra Leone. Sixteen similar villages near the town of Bo were randomly allocated either to remain without nets or to receive treated bednets for all inhabitants, with effect from June 1992. Mosquitoes were sampled using human biting catches on verandas, light-trap catch (beside an occupied untreated bednet), window exit-trap catch and pyrethrum spray collections. During the first year of intervention (June 1992 to July 1993) the treated bednets provided personal protection for people sleeping under them, but had very little impact on densities of An. gambiae collected on human bait. The human blood index (HBI) of An.gambiae was not affected (HBI = 99% in villages with and without nets). An.gambiae parous rates were significantly reduced in all intervention villages, but malaria sporozoite rates fell in only some of the villages. These results are intermediate between those obtained from other projects in Tanzania and Burkina Faso, where treated bednets reduced man-biting, parity and sporozoite rates, versus The Gambia where treated bednets had no significant impact on any of these factors. Possible reasons for these contrasted findings are discussed.     

Anopheles gambiae gonotrophic cycle duration, biting and exiting behaviour unaffected by permethrin-impregnated bednets in The Gambia

Abstract. Permethrin-impregnated bednets protect children against malaria in The Gambia, where Anopheles gambiae complex mosquitoes are the main vectors of malaria. However, no effect has been found on mosquito density, parous rates or sporozoite rates in An.gambiae sensu lato populations; only a reduction in the numbers of mosquitoes resting indoors in rooms with treated bednets. A possible explanation for this paradox is that exposure to treated bednets leads to changed vector behaviour such as a shift in biting time, a diversion to biting outdoors instead of indoors, to biting animals instead of humans, or to increased duration of the gonotrophic cycle. To investigate these possibilities, we observed the biting and exiting behaviour of An.gambiae in ten pairs of villages, in half of which the residents used permethrin-treated bednets. The possible influence of treated bednets on the gonotrophic cycle length was evaluated by mark-release-recapture experiment. No significant difference was found between villages with treated and untreated bednets in the indoor/outdoor ratio of human biting, in mean biting times or in human blood indices of An.gambiae females found resting indoors in the mornings. The proportions of unfed, fed or gravid An.gambiae females collected in exit traps, and the number of females exiting showed no significant differences between rooms with treated and untreated bednets. Indications for a gonotrophic cycle length of 2 days were found. No evidence for any change in duration of the gonotrophic cycle in relation to exposure to treated bednets was found, although the number of recaptures was low in the villages with treated bednets. Since equal numbers of infective An.gambiae were found in villages with treated or untreated bednets, and no changes in mosquito behaviour were detected, we cannot account for how children are protected against malaria by treated bednets. One possibility is that mosquitoes divert to bite other hosts, including adults.     

The campaign against malaria in central western Madagascar: comparison of lambda-cyhalothrin and DDT house spraying. II--Parasitological and clinical study]

For malaria vector control in Madagascar, 10 WP (lambda-cyhalothrin 10% wettable powder) was compared with DDT 75% WP for house-spraying, from November 1997 to September 1998. This study was implemented at the fringe of the malaria epidemic zone, in villages on western slopes of the central highlands, outside the area covered for the past five years by routine DDT house-spraying (OPID). Four types of treatment were compared in different areas: 1) DDT 2 g ai/m2 and 2) lambda-cyhalothrin 30 mg ai/m2 in previously unsprayed villages, 3) no intervention (control); 4) yearly DDT spraying (OPID fifth cycle). To investigate the malariological impact of spraying, cross-sectional surveys of the village populations were performed in each study area at intervals of two months, before and after spraying. In the newly sprayed areas, from December to June, malaria indices decreased by 62% in the ICON area and 44% in the DDT area, whereas in the unsprayed village malaria increased by 32% during the same season. There was a similar decrease in the number of gametocyte carriers in the newly sprayed areas. Active malaria case detection among febrile individuals was performed fortnightly in each village outside the OPID area. Results showed decreased malaria incidence from February (two months post-spraying) in the sprayed villages, despite the rainy season, whereas in the unsprayed area the decline occurred only after the main transmission season. This study demonstrated that, parasitologically as well as entomologically, house-spraying with residual insecticide (DDT or ICON) was an effective method for controlling malaria on the western fringes of the Madagascar highlands epidemic zone. Both products were effective, but ICON had slightly better impact than DDT, i.e. more reduction of malaria indices and of vector longevity, less irritancy of mosquitoes. For best results in this area of transition between stable and unstable malaria, we recommend earlier annual spraying (as soon as November) and extension of the OPID barrier towards western and northern slopes of the Plateau.     

AMA1 and MAEBL are important for Plasmodium falciparum sporozoite infection of the liver

The malaria sporozoite injected by a mosquito migrates to the liver by traversing host cells. The sporozoite also traverses hepatocytes before invading a terminal hepatocyte and developing into exoerythrocytic forms. Hepatocyte infection is critical for parasite development into merozoites that infect erythrocytes, and the sporozoite is thus an important target for antimalarial intervention. Here, we investigated two abundant sporozoite proteins of the most virulent malaria parasite Plasmodium falciparum and show that they play important roles during cell traversal and invasion of human hepatocytes. Incubation of P. falciparum sporozoites with R1 peptide, an inhibitor of apical merozoite antigen 1 (AMA1) that blocks merozoite invasion of erythrocytes, strongly reduced cell traversal activity. Consistent with its inhibitory effect on merozoites, R1 peptide also reduced sporozoite entry into human hepatocytes. The strong but incomplete inhibition prompted us to study the AMA‐like protein, merozoite apical erythrocyte‐binding ligand (MAEBL). MAEBL‐deficient P. falciparum sporozoites were severely attenuated for cell traversal activity and hepatocyte entry in vitro and for liver infection in humanized chimeric liver mice. This study shows that AMA1 and MAEBL are important for P. falciparum sporozoites to perform typical functions necessary for infection of human hepatocytes. These two proteins therefore have important roles during infection at distinct points in the life cycle, including the blood, mosquito, and liver stages.     

Changes in the feeding behaviour of a malaria vector,Anopheles farauti Lav., following use of DDT as a residual spray in houses in the British Solomon Islands Protectorate

• 1 To investigate the failure of DDT to interrupt malaria transmission in parts of the British Solomon Islands Protectorate, regular all-night man-biting catches of malaria vectors were made before and after DDT house spraying on San Cristobal Island.
• 2 Changes were observed in the man-biting behaviour of Anopheles farauti. There was a reduction in the degree of entry into houses and a shifting of the times of peak biting. Whereas before spraying the indoor and outdoor biting cycles differed, after spraying there was no difference although both the indoor and outdoor cycles had altered.
• 3 DDT was found to have a deterrent effect on An.farauti but this effect decreased with time.
• 4 DDT also appears to eliminate a dominant indoor feeding fraction of the farauti population. Following this there can be an increase in an outdoor feeding fraction which can be responsible for a resumption of malaria transmission.
• 5 Biting cycles obtained before spraying are also shown for An.koliensis and An.punctulatus.

     

Experimental hut trials of bednets impregnated with synthetic pyrethroid or organophosphate insecticide for mosquito control in The Gambia

1. Nylon bednets impregnated with different insecticides were evaluated in 1988 against wild adult mosquito populations, mostly Mansonia africana (Theobald) and Anopheles gambiae Giles sensu lato, entering experimental verandah-trap huts in The Gambia. Each bednet had six 10 x 10 cm holes made in the walls to simulate torn conditions and permit female mosquitoes to enter and feed on sleepers. 2. Individual net treatments, determined by gas chromatography of net samples from before and after 12 weeks use of the bednets, were: permethrin 670 +/- 159 and 405 +/- 190 mg/m2 (40% loss), cypermethrin 37 +/- 8 and 16 +/- 9 mg/m2 (57% loss), deltamethrin 10 +/- 7 and 10 +/- 8 mg/m2 (no loss), lambda-cyhalothrin 2.6 +/- 0.9 and 1.6 +/- 0.5 mg/m2 (38% loss), pirimiphos-methyl 4017 +/- 117 and 1160 +/- 319 mg/m2 (71% loss). 3. Washing three times in the traditional manner with local cow-fat soap reduced the initial dosages by about 85% of cypermethrin and lambda-cyhalothrin, 99.8% of pirimiphos-methyl and left no detectable residues of deltamethrin or permethrin. 4. The unwashed permethrin-treated bednet reduced the number of mosquitoes entering a hut by 60% of An.gambiae s.l. and 68% of Mansonia spp. This deterrency was less pronounced with the other insecticides and was lost by washing the bednets. 5. Each insecticide, especially lambda-cyhalothrin and pirimiphosmethyl, caused significant mortality rates of mosquitoes that entered huts with impregnated bednets, and prevented the majority of An. gambiae s.l. and Mansonia females from bloodfeeding. Washing completely removed the efficacy of deltamethrin and permethrin treated bednets, whereas nets treated with cypermethrin, lambda-cyhalothrin or pirimiphos-methyl remained significantly insecticidal after washing. 6. Aerial toxicity from the pirimiphos-methyl treated bednet killed 80% of An.gambiae s.l. confined overnight in the hut at the end of the trial, whereas the pyrethroid-treated bednets gave negligible mortality rates of mosquitoes. 7. Sleepers using the bednets had no medical symptoms significantly associated with any of the treatments. On the contrary, from 216 interviews, 4/10 complaints were associated with the use of untreated nets (P approximately 0.05), perhaps because sleepers were kept awake by mosquitoes and became more aware of any ailments. 8. It is concluded that permethrin tends mainly to deter mosquitoes from house-entry, enhancing personal protection, whereas the other insecticides kill higher proportions of the endophilic mosquitoes, which would give better community protection against malaria transmission.     

Plasmodium vivax pre-erythrocytic vaccines

An estimated 229 million cases of malaria occurred worldwide in 2019. Both, Plasmodium falciparum and P. vivax are responsible for most of the malaria disease burden in the world. Despite difficulties in obtaining an accurate number, the global estimates of cases in 2019 are approximately 229 million of which 2.8% are due to P. vivax, and the total number of malaria deaths are approximately 409 million. Regional elimination or global eradication of malaria will be a difficult task, particularly for P. vivax due to the particular biological features related to the hypnozoite, leading to relapse. Countries that have shown successful episodes of a decrease in P. falciparum malaria, are left with remaining P. vivax malaria cases. This is caused by the mechanism that the parasite has evolved to remain dormant in the liver forming hypnozoites. Furthermore, while clinical trials of vaccines against P. falciparum are making fast progress, a very different picture is seen with P. vivax, where only few candidates are currently active in clinical trials. We discuss the challenge that represent the hypnozoite for P. vivax vaccine development, the potential of Controlled Human Malaria Challenges (CHMI) and the leading vaccine candidates assessed in clinical trials.     

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.     

Movement of Anopheles gambiae s.l. malaria vectors between villages in The Gambia

ovement of mosquitoes belonging to the Anopheles gambiae complex (mixed wild populations of An.arabiensis, An.gambiae and An.melas ) between three neighbouring rural villages in The Gambia was investigated by mark-release-recapture. A total of 12,872 mosquitoes were collected in bednets, marked with a magenta fluorescent powder and released over a 15-day period in one of the villages. A further 15,507 mosquitoes were collected in exit traps, marked with a yellow powder and released over the same period. Mosquitoes were captured daily in all three villages using pyrethrum spray catches, as well as bednet and exit trap catches. The catching period extended for 6 days after the last day of release.
Of the mosquitoes released, 372 (1.3%) were recaptured 2–21 days later. Of these recaptures, 272 were caught in the release village, and 98 were caught in other villages situated 1–1.4 km away. The 'movement index' between villages was calculated as 17.2% (12.2–22.4% confidence limits) for mosquitoes released after feeding and 20.1% (14.7–25.3%) for those released unfed.
These results suggest that movement of mosquitoes between neighbouring villages in The Gambia seriously affects the entomological evaluation of pyrethroid-impregnated bednet programmes in areas where treated and untreated villages are interspersed.     

Allozyme analysis reveals six species within the Anopheles punctulatus complex of mosquitoes in Papua New Guinea

Abstract. Among samples collected from nineteen localities in Papua New Guinea, we have identified six species within the Anopheles punctulatus complex of mosquitoes, by means of cellulose acetate allozyme electrophoresis. An.punctulatus Dönitz sensu stricto was collected from seven villages in the Madang area and from Buksak, Sausi Mission and an area 18 km SW of Tari; An.koliensis Owen from eight villages in the Madang area, from Popondetta and Brown River near Karema; and An.farauti No. 1 from ten coastal areas including Madang, Lorengau, Popondetta, Port Moresby, Rabaul and Wewak. Three newly recognized species, reported here for the first time, are designated as An.farauti No. 4 from Gonoa and Hudini, Madang area; An.farauti No. 5 from Ketarabo near Goroka; and An.farauti No. 6 from Hiwanda near Tari. Three other known members of the complex, An.clowi Rozeboom & Knight, An.farauti No. 2 (Bryan, 1973) and An.farauti No. 3 (Mahon & Meithke, 1982) were not detected in Papua New Guinea. Problems arising with morphological characters for the identification of species in this group are discussed.     

Malaria vectors of Papua New Guinea

Understanding malaria transmission in Papua New Guinea (PNG) requires exact knowledge of which Anopheles species are transmitting malaria and is complicated by the cryptic species status of many of these mosquitoes. To identify the malaria vectors in PNG we studied Anopheles specimens from 232 collection localities around human habitation throughout PNG (using CO₂ baited light traps and human bait collections). A total of 22,970 mosquitoes were individually assessed using a Plasmodium sporozoite enzyme-linked immunosorbent assay to identify Plasmodiumfalciparum, Plasmodiumvivax and Plasmodiummalariae circumsporozoite proteins. All mosquitoes were identified to species by morphology and/or PCR. Based on distribution, abundance and their ability to develop sporozoites, we identified five species as major vectors of malaria in PNG. These included: Anophelesfarauti, Anopheleshinesorum (incriminated here, to our knowledge, for the first time), Anophelesfarauti 4, Anopheleskoliensis and Anophelespunctulatus. Anopheleslongirostris and Anophelesbancroftii were also incriminated in this study. Surprisingly, An. longirostris showed a high incidence of infections in some areas. A newly identified taxon within the Punctulatus Group, tentatively called An. farauti 8, was also found positive for circumsporozoite protein. These latter three species, together with Anopheleskarwari and Anophelessubpictus, incriminated in other studies, appear to be only minor vectors, while Anophelesfarauti 6 appears to be the major vector in the highland river valleys (>1500 m above sea level). The nine remaining Anopheles species found in PNG have been little studied and their bionomics are unknown; most appear to be uncommon with limited distribution and their possible role in malaria transmission has yet to be determined.     

Long-term use of permethrin-impregnated nets does not increase Anopheles gambiae permethrin tolerance

Abstract. Previous use of permethrin-impregnated bednets (mosquito nets) and curtains in four Kenyan villages for one year, 1990-91, raised the permethrin LT₅₀ of Anopheles gambiae to 2.4-fold above its baseline value, designated permethrin tolerance (PT), as measured by exposure to 0.25% permethrin-impregnated papers in W.H.O. test-kits. During 1992-93, with ongoing use of permethrin-impregnated nets and curtains, PT regressed slightly compared with the contemporary susceptibility level of An. gambiae from non-intervention villages, to 1.8-fold in 1992 and only 1.6-fold in 1993. Thus the selection pressure of impregnated nets for PT in Angambiae appears to be minimal in our study villages, although the impact of permethrin was demonstrated by a significantly lower parous-rate of Angambiae females in the intervention (63–66%) than in non-intervention (79%) villages, and by reduced malaria transmission (reported elsewhere).
In a selected stock of An. gambiae from the study area, PT did not affect the susceptibility to deltamethrin, fenitrothion, propoxur or DDT. Bioassays described herein provide easy procedures for field-monitoring of mosquito susceptibility/tolerance/resistance to insecticides used for net impregnation in operational programmes.     

Experimental hut trials of permethrin-impregnated mosquito nets and eave curtains against malaria vectors in Tanzania

Permethrin impregnated netting was tested against Tanzanian populations of Anopheles arabiensis Patton, An.gambiae Giles and An.funestus Giles in experimental huts fitted with traps to catch samples of the mosquitoes existing during the night. Treated bednets killed some mosquitoes and increased the tendency of survivors to exit during the night. Treated cotton did not perform so well as treated nylon bednets. An impregnated bednet in which holes had been cut, to simulate a torn net, reduced the number of mosquitoes which fed and survived approximately as well as an intact untreated net. Treated curtains around the eaves of experimental huts did not perform so well as bednets but caused considerable reductions in the number of mosquitoes which fed and survived. However, there was no such effect when treated netting was placed around the eaves of a dwelling house. When one child slept under a treated net and another slept outside the net in the same hut, the number of bites on the latter child was less than if neither child had been under a net. Various aspects of the applicability of permethrin impregnated nets on a community basis are discussed.