Armigeres subalbatus colonization of damaged pit latrines: a nuisance and potential health risk to residents of resettlement villages in Laos.

During the resettlement of 6500 persons living around the Nam Theun 2 hydroelectric project in Laos, more than 1200 pour-flush latrines were constructed. To assess the role of these latrines as productive larval habitats for mosquitoes, entomological investigations using Centers for Disease Control (CDC) light traps, visual inspection and emergence trapping were carried out in over 300 latrines during the rainy seasons of 2008-2010. Armigeres subalbatus (Diptera: Culicidae) were nine times more likely to be found in latrines (mean catch: 3.09) than in adjacent bedrooms (mean catch: 0.37) [odds ratio (OR) 9.08, 95% confidence interval (CI) 6.74-15.11] and mosquitoes were active in and around 59% of latrines at dusk. Armigeres subalbatus was strongly associated with latrines with damaged or improperly sealed septic tank covers (OR 5.44, 95% CI 2.02-14.67; P < 0.001). Armigeres subalbatus is a nuisance biter and a putative vector of Japanese encephalitis and dengue viruses. Dengue virus serotype 3 was identified from a single pool of non-blood-fed female A. subalbatus using reverse transcription polymerase chain reaction. Maintaining a good seal around septic tanks by covering them with a layer of soil is a simple intervention to block mosquito exit/entry and contribute to vector control in resettlement villages. The scale-up of this simple, cheap intervention would have global impact in preventing the colonization of septic tanks by nuisance biting and disease-transmitting mosquitoes.

Preliminary studies developing methods for the control of Chrysomya putoria, the African latrine fly, in pit latrines in The Gambia.

OBJECTIVE: To explore ways of controlling Chrysomya putoria, the African latrine fly, in pit latrines. As pit latrines are a major source of these flies, eliminating these important breeding sites is likely to reduce village fly populations, and may reduce the spread of diarrhoeal pathogens. METHODS: We treated 24 latrines in a Gambian village: six each with (i) pyriproxyfen, an insect juvenile hormone mimic formulated as Sumilarv(®) 0.5 G, a 0.5% pyriproxyfen granule, (ii) expanded polystyrene beads (EPB), (iii) local soap or (iv) no treatment as controls. Flies were collected using exit traps placed over the drop holes, weekly for five weeks. In a separate study, we tested whether latrines also function as efficient flytraps using the faecal odours as attractants. We constructed six pit latrines each with a built-in flytrap and tested their catching efficiency compared to six fish-baited box traps positioned 10 m from the latrine. Focus group discussions conducted afterwards assessed the acceptability of the flytrap latrines. RESULTS: Numbers of emerging C. putoria were reduced by 96.0% (95% CIs: 94.5-97.2%) 4-5 weeks after treatment with pyriproxyfen; by 64.2% (95% CIs: 51.8-73.5%) after treatment with local soap; by 41.3% (95% CIs = 24.0-54.7%) after treatment with EPB 3-5 weeks after treatment. Flytraps placed on latrines collected C. putoria and were deemed acceptable to local communities. CONCLUSIONS: Sumilarv 0.5 G shows promise as a chemical control agent, whilst odour-baited latrine traps may prove a useful method of non-chemical fly control. Both methods warrant further development to reduce fly production from pit latrines. A combination of interventions may prove effective for the control of latrine flies and the diseases they transmit.

Habitat associations, reproduction and diet of the Guinean tilapia Tilapia guineensis of the Gambia River floodplains.

The ecology of the Guinean tilapia Tilapia guineensis a dominant species of the lower Gambia River floodplains and an important food source in parts of West Africa was studied to better understand the threat posed from construction of a barrage across the river. The catch per unit effort (CPUE) of T. guineensis was positively associated with conductivity and dissolved oxygen and negatively associated with water depth and the % vegetation cover. Diet studies indicated that the T. guineensis is primarily an iliophage. The peak of reproduction was at the beginning of the rainy season. CPUE peaked in May, just before the first rains, and subsequently declined, probably reflecting movement into newly flooded habitat. Median size at maturity was 11.6 cm total length, L(T), for females and 12.5 cm L(T) for males. L(T)-frequency analysis indicated several juvenile cohorts as well as very young fish on the floodplains each month, suggesting continuation of reproduction throughout the rainy season. The impending construction of a hydroelectric dam on the Gambia River is likely to affect T. guineensis negatively through anticipated changes in the hydrology of the river.

Mapping the ranges and relative abundance of the two principal African malaria vectors, Anopheles gambiae sensu stricto and An. arabiensis, using climate data.

Members of the Anopheles gambiae complex are major malaria vectors in Africa. We tested the hypothesis that the range and relative abundance of the two major vectors in the complex, An. gambiae sensu stricto and An. arabiensis, could be defined by climate. Climate was characterized at mosquito survey sites by extracting data for each location from climate surfaces using a Geographical Information System. Annual precipitation, together with annual and wet season temperature, defined the ranges of both vectors and were used to map suitable climate zones. Using data from West Africa, we found that where the species were sympatric, An. gambiae s.s. predominated in saturated environments, and An. arabiensis was more common in sites subject to desiccation (r2 = 0.875, p < 0.001). We used the nonlinear equation that best described this relationship to map habitat suitability across Africa. This simple model predicted accurately the relative abundance of both vectors in Tanzania (rs = 0.745, p = 0.002), where species composition is highly variable. The combined maps of species' range and relative abundance showed very good agreement with published maps. This technique represents a new approach to mapping the distribution of malaria vectors over large areas and may facilitate species-specific vector control activities.

Safety of the insect repellent N,N-diethyl-M-toluamide (DEET) in pregnancy.

The safety of daily application of N, N-diethyl-m-toluamide (DEET) (1.7 g of DEET/day) in the second and third trimesters of pregnancy was assessed as part of a double-blind, randomized, therapeutic trial of insect repellents for the prevention of malaria in pregnancy (n = 897). No adverse neurologic, gastrointestinal, or dermatologic effects were observed for women who applied a median total dose of 214.2 g of DEET per pregnancy (range = 0-345.1 g). DEET crossed the placenta and was detected in 8% (95% confidence interval = 2.6-18.2) of cord blood samples from a randomly selected subgroup of DEET users (n = 50). No adverse effects on survival, growth, or development at birth, or at one year, were found. This is the first study to document the safety of DEET applied regularly in the second and third trimesters of pregnancy. The results suggest that the risk of DEET accumulating in the fetus is low and that DEET is safe to use in later pregnancy.

The effect of different types of skin surfaces on the transmission of Brugia pahangi infective larvae by the mosquito Aedes aegypti.

The transmission of Brugia pahangi from Aedes aegypti into Meriones unguiculatus was compared for four different skin surfaces (HAIRY, CLOTH, CLEAN, GREASY). Unshaven jirds reduced the feeding avidity of infective mosquitoes. The loss of larvae from infective mosquitoes was greatest when these insects fed on "exposed" skin surfaces. Significantly fewer infective larvae penetrated the host when infective mosquitoes fed on a jird through a thin layer of cloth.

Local-scale variation in malaria infection amongst rural Gambian children estimated by satellite remote sensing.

We investigated local-scale variation in malaria transmission and infection in children within a continuous landscape by retrospective spatial analysis of entomological and clinical data collected during 1988 and 1989 in The Gambia, West Africa. Parasite prevalence was negatively correlated with vector abundance and exposure to malaria parasites in 10 villages where entomological surveillance had been carried out. Variation in bednet use did not explain this finding. Mosquito-breeding habitat was retrospectively mapped using 20-m spatial resolution multispectral SPOT satellite imagery from 1988. From these data we estimated by linear regression the risk of exposure to malaria parasites in 26 villages where clinical surveys of children had been made. As exposure increased, so did parasite prevalence; but at higher levels of exposure, parasite prevalence declined. Our findings demonstrate marked differences in exposure to malaria in villages over distances of less than 2 km from mosquito breeding sites and suggest that there are also large differences in immunity between neighbouring settlements.

Mapping and estimating the population at risk from lymphatic filariasis in Africa.

Lymphatic filariasis remains a major public health problem in Africa and is 1 of the World Health Organization's 6 diseases targeted for global eradication. However, no detailed maps of the geographical distribution of this disease exist, making it difficult to target control activities and quantify the population at risk. We hypothesized that the distribution lymphatic filariasis is governed by climate. The climate at sites in Africa where surveys for lymphatic filariasis had taken place was characterized using computerized climate surfaces. Logistic regression analysis of the climate variables predicted with 76% accuracy whether sites had microfilaraemic patients or not. We used the logistic equation in a geographical information system to map risk of lymphatic filariasis infection across Africa, which compared favourably with expert opinion. Further validation with a quasi-independent data set showed that the model predicted correctly 88% of infected sites. A similar procedure was used to map risk of microfilaraemia in Egypt, where the dominant vector species differs from those in sub-Saharan Africa. By overlaying risk maps on a 1990 population grid, and adjusting for recent population increases, we estimate that around 420 million people will be exposed to this infection in Africa in the year 2000. This approach could be used to produce a sampling frame, based on estimated risk of microfilaraemia, for conducting filariasis surveys in countries that lack accurate distribution maps and thus save on costs.

The effect of humidity on the transmission of Brugia pahangi infective larvae to mammalian hosts by Aedes aegypti.

The transmission of Brugia pahangi by Aedes aegypti to the mammalian host was compared at low and high humidity. There was no statistical difference between the number of infected mosquitoes feeding or the egress of infective larvae from these mosquitoes at high or low humidity. The penetration of the host by the infective larvae was however significantly greater (p less than 0.05) at high than at a low humidity.

A trial of bed nets (mosquito nets) as a malaria control strategy in a rural area of The Gambia, West Africa.

An intervention trial was undertaken in a rural area of The Gambia to assess the impact on malaria morbidity of the use of bed nets. Bed nets were allocated at random among a group of 16 Fula hamlets, where they were previously rarely used. The incidence of febrile episodes with associated malaria parasitaemias throughout the rainy season and the prevalence of splenomegaly and parasitaemia at the end of the rainy season were determined in 233 children aged 1-9 years who slept under bed nets and in 163 children who did not. Bed nets were used correctly by the children in the study cohort, but direct observations showed that a significant number of children left their nets for a period during the night. There was no significant difference in the incidence of clinical attacks of malaria or in any other malariometric measurement between the 2 groups. Thus, bed nets were not effective in reducing malaria morbidity in this group of children. The apparent protection from bed nets demonstrated in previous retrospective surveys may have been due to an increased number of infective bites being received by exposed individuals sleeping close to users of bed nets.

Permethrin-treated bed nets (mosquito nets) prevent malaria in Gambian children.

The incidence of clinical attacks of malaria was significantly less in Gambian children aged 1-9 years who slept in villages where all the bed nets (mosquito nets) were treated with permethrin than in children who slept in control villages with placebo-treated nets. Significant differences in changes in spleen size and in packed cell volume were also observed between the 2 groups during the course of a rainy season. No side effect was noted. Treatment of bed nets with insecticide is a form of malaria control that is well suited to community participation and can readily be incorporated into primary health care programmes. Insecticide-treated nets may be more effective in areas of seasonal or low intensity transmission than in areas with heavy perennial challenge.

Irrigated crop production is associated with less malaria than traditional agricultural practices in Tanzania.

There is concern that crop irrigation that results in increased numbers of vector mosquitoes will lead to a rise in malaria in local communities. We evaluated the level of malaria experienced in 3 communities in northern Tanzania with different agricultural practices: rice irrigation, sugar-cane irrigation and traditional maize cultivation. Five cross-sectional surveys were used to measure the prevalence of infection with falciparum malaria in 1-4 years old children in each community over a period of 12 months. Active case detection was also carried out to record clinical episodes of malaria during the study period. Information on antimalarial measures was also recorded. Results from the cross-sectional surveys showed that the overall prevalence of malaria parasites was less near the rice irrigation (12.5%) and sugar-cane (16.9%) schemes than the savannah village (29.4%). There were also significantly fewer clinical episodes of malaria in the rice village (15 cases/1000 child-weeks at risk [cwar]) than either the sugar-cane (36 cases/1000 cwar) or savannah (40 cases/1000 cwar) villages. Overall, rice irrigation was associated with less malaria than alternative agricultural practices, despite the considerable numbers of vectors produced in the paddies. This finding supports other studies that indicate that irrigation in much of sub-Saharan Africa will not lead to increased malaria. Nonetheless, African governments planning irrigation projects need effective policies to encourage local communities to use personal protection measures, such as insecticide-treated bednets, and to ensure that these communities have access to effective antimalarial drugs and efficient health services.

Short-range attractiveness of pregnant women to Anopheles gambiae mosquitoes.

Malaria is a major cause of illness and an indirect cause of mortality in pregnant women. It can also cause stillbirths and low-birthweight babies. We have shown previously that pregnant women attracted twice as many Anopheles gambiae mosquitoes, the principal African malaria vector, as their non-pregnant counterparts over distances of about 15 m. In the current study (in 1998/99) we compared the short-range attractiveness of both pregnant and non-pregnant women sleeping under untreated bednets in Gambian villages. First, we measured the rate of mosquito entry under bednets and, second, we calculated the proportion of mosquitoes biting mothers under each bednet compared to their children. The feeding preference of An. gambiae collected under nets was determined by DNA fingerprinting blood samples from human subjects sleeping under each bednet and comparing these to fingerprints obtained from mosquito bloodmeals. Pregnant women were more attractive to An. gambiae mosquitoes than non-pregnant women under an untreated bednet. The number of mosquitoes entering bednets each night was 1.7-4.5 times higher in the pregnant group (P = 0.02) and pregnant women also received a higher proportion of bites under the bednets than did non-pregnant women (70% vs 52%, P = 0.001). This study clearly demonstrates that pregnant women are more exposed to malaria parasites than other women, which contributes to the greater vulnerability of pregnant women to malaria.

Effect of topography on the risk of malaria infection in the Usambara Mountains, Tanzania.

We investigated whether the risk of infection with malaria parasites was related to topography in the Usambara Mountains, Tanzania. Clinical surveys were carried out in seven villages, situated at altitudes from 300 m to 1650 m. Each village was mapped and incorporated into a Digital Terrain Model. Univariate analysis showed that the risk of splenomegaly declined with increasing altitude and with decreasing potential for water to accumulate. Logistic regression showed that altitude alone could correctly predict 73% of households where an occupant had an enlarged spleen or not. The inclusion of land where water is likely to accumulate within 400 m of each household increased the accuracy of the overall model slightly to 76%, but significantly improved predictions between 1000 m and 1200 m, where malaria is unstable, and likely to be epidemic. This novel approach illustrates how topography could help identify local areas prone to epidemics in the African highlands.

Do untreated bednets protect against malaria?

Bednets are thought to offer little, if any, protection against malaria, unless treated with insecticide. There is also concern that the use of untreated nets will cause people sleeping without nets to receive more mosquito bites, and thus increase the malaria risk for other community members. Regular retreatment of nets is therefore viewed as critical for malaria control. However, despite good uptake of nets, many control programmes in Africa have reported low re-treatment rates. We investigated whether untreated bednets had any protective benefit (in October and November 1996) in The Gambia where nets, although widely used, are mostly untreated. Cross-sectional prevalence surveys were carried out in 48 villages and the risk of malaria parasitaemia was compared in young children sleeping with or without nets. Use of an untreated bednet in good condition was associated with a significantly lower prevalence of Plasmodium falciparum infection (51% protection [95% CI 34-64%], P < 0.001). This finding was only partly explained by differences in wealth between households, and children in the poorest households benefited most from sleeping under an untreated net (62% protection [14-83%], P = 0.018). There was no evidence that mosquitoes were diverted to feed on children sleeping without nets. These findings suggest that an untreated net, provided it is in relatively good condition, can protect against malaria. Control programmes should target the poorest households as they may have the most to gain from using nets.

Transmission ecology of the fly Musca sorbens, a putative vector of trachoma.

Recent evidence suggests that eye-seeking flies are important trachoma vectors. We conducted a series of investigations to identify which species of synanthropic flies are potential vector(s) of this blinding disease in The Gambia. Several species of fly were caught in fish-baited attractant traps placed in villages throughout the year (1997/98) but only 2 species, Musca sorbens and M. domestica, were caught from the eyes of children. M. sorbens comprised < 10% of the total number of flies caught with attractant traps but was responsible for > 90% of fly-eye contacts, the remainder were made by M. domestica. All fly species were more numerous in the wet season than the dry season. Eyes of young children are considered to be the main reservoir of Chlamydia trachomatis, the causative agent of trachoma. Collections of eye-seeking flies from children showed frequent fly-eye contacts (median [interquartile range], 3 [1.5-7] every 15 min). Children with potentially infective ocular or nasal discharge had twice as many fly-eye contacts than children with no discharge (P < 0.001). There was no difference in exposure to fly-eye contacts if a child sat inside or outside a house (P = 0.273). Female flies were more commonly caught from eyes than male (P < 0.001). The presence of Chlamydia DNA was demonstrated by PCR on 2 of 395 flies caught from the eyes of children with a current active trachoma infection. Both positive flies were M. sorbens, one male and the other female. Further elucidation of M. sorbens behavioural ecology and the development of sustainable strategies to control these flies should be a priority. It is likely that M. sorbens is the principal insect vector of trachoma in The Gambia.

Improved method for distinguishing the human source of mosquito blood meals between close family members.

We have developed a simple and relatively cheap method to distinguish the origin of mosquito blood meals between close family members, effective for both laboratory and field samples. Each blood meal was squashed on to filter paper and eluted overnight with 0.5 mL phosphate-buffered saline. Deoxyribonucleic acid (DNA) was extracted using a chemical matrix (Insta-gene) which bound to everything from the blood meal except DNA, which remained in the supernatant. Following extractions, reference DNA samples taken directly from finger-prick blood of human subjects and those from blood meals of unknown origin were amplified with human microsatellite markers using a thermal cycler. Polymerase chain reaction products were then run on an ABI gel (Automated Biosystems) to obtain a genotype for each sample. The DNA from each mosquito blood meal was then matched to an individual host. With laboratory samples, human DNA which had been extracted from mosquito blood meals up to 12 h after feeding could be used. One important application of this method will be to identify which members of a community are most at risk from vector-borne diseases. It also has numerous potential applications in studies of insect biting behaviour in both human and veterinary science.

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 2. Mortality and morbidity from malaria in the study area.

Background data on child mortality and morbidity from malaria were obtained in a new study area in the centre of The Gambia, south of the river, chosen as the site for a malaria intervention trial. Infant and child mortality rates were 120 and 41 per 1000 respectively. Results obtained using post-mortem questionnaires suggested that malaria was an uncommon cause of death in children under the age of one year but responsible for about 40% of deaths in children aged 1-4 years. Ninety-two percent of deaths attributed to malaria occurred during or immediately after the rainy season. Parasite and spleen rates in children aged 1-5 years at the end of the malaria transmission season were 66% and 64% respectively. Malariometric indices were similar in primary health care (PHC) villages, selected as sites for an intervention with insecticide-treated bed nets and targeted chemoprophylaxis, and in smaller, non-PHC, control villages.

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 3. Entomological characteristics of the study area.

Baseline entomological surveillance was carried out in a rural area of The Gambia during the rainy season in 1988, one year before the implementation of a malaria control programme using insecticide-impregnated nets and targeted chemoprophylaxis in villages with a primary health care (PHC) system. Mosquito collections took place in 6 pairs of settlements each with untreated bed nets; within each pair there was a large PHC village with a resident village health worker (VHW) and traditional birth attendant (TBA) and a smaller non-PHC village without either a VHW or a TBA. The most common vectors in the study area were Anopheles gambiae sensu stricto and, to a lesser extent, An. arabiensis. These mosquitoes were found in appreciable numbers for at least 4 months of the year (geometric mean/bedroom/night = 32.5, 95% confidence interval 18.2-57.3). Numbers of mosquitoes collected in PHC villages or non-PHC villages were not significantly different. Greater numbers of mosquitoes were found in villages closer to the River Gambia than in those further away. Evidence for DDT resistance due to elevated glutathione S-transferase activity was found in one of the 12 villages, but there was no evidence of resistance to organophosphate or carbamate insecticides as suggested by the low esterase levels and carbamate sensitive acetylcholinesterase.

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 4. Perceptions of the causes of malaria and of its treatment and prevention in the study area.

Perceptions of the causes of malaria, its treatment and prevention were studied among 996 adults, selected randomly from 73 villages and hamlets in a rural area of The Gambia. Structured questionnaires and other interview techniques were used for data collection. Malaria has no specific name in the study area; it is referred to commonly as Fula kajewo (Fula fever). Only 28% of the respondents knew that mosquitoes transmitted malaria. However, most people believed correctly that August to October was the main malaria season. Eighty-six per cent of the subjects were bed net users. The majority of nets were produced locally, usually white in colour and made of sheeting fabrics. Usage of nets was correlated with ethnic group, age and polygamy but not with education, income, occupation or ownership of certain items which indicate high social status. Analysis of expenditure on mosquito coils indicated that non-users of nets spent 43% more on coils than did users. Bed nets have been used for a long time in the study area; 98% of users saw their parents using them during their childhood.

PIP: In the Gambia, malaria specialists at the Medical Research Council Laboratories oversaw interviews of 996 men and women from, 73 villages and hamlets on the south bank of the River Gambia to learn their perceptions of the causes of malaria and of its treatment and prevention. No specific local term for malaria existed. The most common term used by all ethnic groups, except the Fulas, was Fula kejewo (Fula fever). Just 28% of all adults knew that mosquitoes transmit malaria. Men were more likely to know the real cause of malaria than women (p .001). Knowledge about the cause of malaria increased with education p =.01). Most people (90%) knew that malaria transmission occurs most often during August-October. 86% of adults and 81% of their children less than 10 years old used bed nets. Girls were more likely to sleep under bed nets than boys (p = .005). Local tailors made most bed nets with either second hand or new fabrics. They tended to be made of sheeting fabrics and white in color. Most adults were willing to treat their bed nets with the insecticide permethrin to protect against mosquitoes. Ethnic group (Fulas less likely, p .001), age (older people more likely, p .001), and polygamy (p = .002) were correlated with bed net use. Education, income, and occupation or ownership of items indicating high social status were not correlated with bed net use. Nonusers of bed nets spent 43% more on mosquito coils, used to smoke out mosquitoes from rooms before going to sleep, than did users (p = .001). Almost all adults (98%) remembers their parents using bed nets when they were children. The bed nets users who were at least 70 years old said that the Mandinkas mainly used bed nets. These findings should prove useful to the government as it develops a national impregnated bed net program to control malaria in rural areas.