Modelling the effect of temperature on transmission of dengue.

The main entomological parameters involved in the rate of dengue virus transmission include the longevity of female mosquitoes, the time interval between bites and the extrinsic incubation period of the virus. Field and laboratory data provide estimates for these parameters, but their interactions with other factors (e.g. host population density and environmental parameters) make their integration into a transmission model quite complex. To estimate the impact of these parameters on transmission, we developed a model of virus transmission by a vector population which predicts the number of potentially infective bites under a range of temperatures and entomological parameters, including the daily survival rate of females, the interval between bites and the extrinsic incubation period. Results show that in a stable population, an increase in mosquito longevity disproportionately enhances the number of potential transmissions (e.g. by as much as five times when the survival rate rises from 0.80 to 0.95). Halving the length of the biting interval with a 10- degrees C rise in temperature increases the transmission rate by at least 2.4 times. Accordingly, the model can predict changes in dengue transmission associated with short-term variation in seasonal temperature and also with potentially long-lasting increases in global temperatures.

Reducing costs and operational constraints of dengue vector control by targeting productive breeding places: a multi-country non-inferiority cluster randomized trial.

OBJECTIVES: To test the non-inferiority hypothesis that a vector control approach targeting only the most productive water container types gives the same or greater reduction of the vector population as a non-targeted approach in different ecological settings and to analyse whether the targeted intervention is less costly. METHODS: Cluster randomized trial in eight study sites (Venezuela, Mexico, Peru, Kenya, Thailand, Myanmar, Vietnam, Philippines), with each study area divided into 18-20 clusters (sectors or neighbourhoods) of approximately 50-100 households each. Using a baseline pupal-demographic survey, the most productive container types were identified which produced >or=55% of all Ae. aegypti pupae. Clusters were then paired based on similar pupae per person indices. One cluster from each pair was randomly allocated to receive the targeted vector control intervention; the other received the 'blanket' (non-targeted) intervention attempting to reach all water holding containers. RESULTS: The pupal-demographic baseline survey showed a large variation of productive container types across all study sites. In four sites the vector control interventions in both study arms were insecticidal and in the other four sites, non-insecticidal (environmental management and/or biological control methods). Both approaches were associated with a reduction of outcome indicators in the targeted and non-targeted intervention arm of the six study sites where the follow up study was conducted (PPI, Pupae per Person Index and BI, Breteau Index). Targeted interventions were as effective as non-targeted ones in terms of PPI. The direct costs per house reached were lower in targeted intervention clusters than in non-targeted intervention clusters with only one exception, where the targeted intervention was delivered through staff-intensive social mobilization. CONCLUSIONS: Targeting only the most productive water container types (roughly half of all water holding container types) was as effective in lowering entomological indices as targeting all water holding containers at lower implementation costs. Further research is required to establish the most efficacious method or combination of methods for targeted dengue vector interventions.

Assessment of a new strategy, based on Aedes aegypti (L.) pupal productivity, for the surveillance and control of dengue transmission in Thailand.

In the countries where the disease is endemic, control of dengue is mainly based on the elimination or treatment of the water-filled containers where the main vector, Aedes aegypti, breeds, in interventions usually reliant on community participation. Although such control activities must be continuous, since vector eradication appears impossible, it should be possible to reduce the incidence of dengue significantly, in a cost-effective manner, by targeting only those types of containers in which large numbers of Ae. aegypti are produced. This strategy is now recommended by the World Health Organization, although it depends on the most productive types of container being carefully identified, in each endemic region. In Thailand, exhaustive surveys of 3125 wet containers in 240 houses in either an urban area (100-120 houses) or a rural area (120 houses) were conducted during a rainy and a dry season in 2004-2005. Indices based on the numbers of Ae. aegypti pupae observed were found to correlate with the 'classical' entomological indices that are based on all of the immature stages of the vector. Overall, 2.3 and 0.8 Ae. aegypti pupae were observed per person in the rural and urban areas, respectively. Although adult female Ae. aegypti laid eggs in all 10 types of wet container that were identified, large water-storage containers produced the majority of the pupae, especially at the end of the dry season (when such containers accounted for 90% of the pupae detected in the rural area and 60% of those in the urban area). Since these containers are large, easy to reach and account for, <50% of all wet containers, it should be relatively easy and quick to treat them with larvicide or to cover them. If even such targeted treatment is to be sustainable, however, it will have to be integrated, as one of several activities in which the at-risk communities are encouraged to participate.

Sample sizes for identifying the key types of container occupied by dengue-vector pupae: the use of entropy in analyses of compositional data.

A method has been developed for estimating the sample sizes needed to identify categories that comprise a large proportion of a compositional data-set. The method is to be used in the design of surveys of mosquito pupae, for identifying the key container types from which the majority of adult dengue vectors emerge. Although a finite-population correction was devised for estimating the mean of a negative binomial distribution, other complications of parametric approaches make them unlikely to yield methods simple enough to be practically applicable. The Shannon-Wiener index was therefore investigated as a more useful alternative, at the cost of theoretical generalizability, in an approach based on re-sampling methods in conjunction with the use of entropy. This index can be used to summarize the degree to which pupae are either concentrated in a few container types, or dispersed among many. An empirical relationship between the index and the repeatability of surveys of differing sample sizes was observed. A step-wise rule, based on the entropy of the cumulative data, was devised for determining the sample size, in terms of the number of houses positive for pupae, at which a pupal survey might reasonably be stopped.

[An experimental model to analyse the characteristics of a strain of Culex pipiens and its sensitivity to insecticides]. / Un modèle expérimental d'analyse de caractères d'une souche de Culex pipiens et de sa sensibilité aux insecticides.

This study, based on phenotype of green larvae colour, has enabled the selection of a genetically distinct line of Culex pipiens pipiens. The four loci under study were found to be homozygous to the 42nd generation. These loci were associated with an increased sensitivity to organochlorides in the progeny when compared to the parent strain. This observation, with possible expansion to other species, has potential practical applications in the identification of susceptible insect populations in insecticide campaigns.

[Entomologic study on the transmission of onchocerciasis in a forest-savanna transition area of Cameroon]. / Etude entomologique sur la transmission de l'onchocercose dans une zone de transition forêt-savane du Cameroun.

An entomological study was conducted in the basin area of middle Sanaga (Cameroon) in order to document the pattern of onchocerciasis transmission in a region where a high prevalence of infection had been recorded in villages located 30 km from the main rivers. The main vector of O. volvulus was found to be S. squamosum s.s. No breeding site was found in the small tributaries of the Sanaga and Mbam Rivers, and the dispersal of S. damnosum s.l. in the area thus appeared to be particularly high. The highest blackfly population densities were recorded during the long rainy season, and a second peak of density occurred along the Mbam River during the short rainy season. A seasonal variation in dispersal patterns was found at three of the four transects studied. The transmission of Onchocerca volvulus in the area occurred principally between January and May (i.e. at the end of the long dry season and the beginning of the short rainy season). Vector control operations might well reinforce the effect of ivermectin distributions in this onchocerciasis focus.

Impact of treatments with Bacillus sphaericus on Anopheles populations and the transmission of malaria in Maroua, a large city in a savannah region of Cameroon.

Simultaneously with a control of breeding sites primarily for Culex quinquefasciatus and secondarily for anophelines with Bacillus sphaericus in the town of Maroua (120,000 inhabitants) in North Cameroon, a survey of anopheline populations and of transmission rates of malaria was performed. Monthly night catches in 8 districts of the town emphasized the relation between the biting rate by Anopheles in the districts and two main factors. One factor was the distance of a district from the breeding sites, i.e., natural flooded areas along the periphery of the town or artificial breeding sites (ditches, puddles) filled with rain water during the rainy season and with water from the water network throughout the year. The second factor was the density of the habitation that reduced dispersal of female mosquitoes from the breeding sites and the risk for inhabitants to be injected because of scattered bites. The treatment with B. sphaericus was followed by a delay (2 months) in the beginning of the transmission period and a decrease in the incidence of malaria cases studied in a health facility of the town. It thus seems to be possible to reduce malaria transmission by applying B. sphaericus to the breeding sites, but this requires a good knowledge of the location and dynamics of breeding sites and an improved formulation of the pesticide.

[Geographic determinants of onchocerciasis transmission in a forest-savannah transition zone: an example of 2 villages of the Mbam focus (central region, Cameroon)]. / Les déterminants géographiques de la transmission de l'onchocercose en zone de transition forêt-savane: l'exemple de deux villages du foyer du Mbam (région du Centre, Cameroun).

Mbam, a focus of onchocerciasis outbreaks, is located in the forest/savannah transition zone of Cameroon. Transmission of the disease in this area is similar to that sometimes observed in West Africa. Geographical factors affecting transmission must be identified to determine the areas for priority action. The locations of breeding sites for black flies (the vector of the disease) and factors favorable to their dispersion, the social behavior of these population and the way that she occupy space, must be investigated to identify areas in which the flies and man are in close contact. This geographical study shows that the closest contact between men and black flies occurs in cocoa plantations, due to the proximity of the breeding sites. The areas of closet contact for women and children are unknown, but their identification is important because women and children may be bitten by the flies during their daily activities.

Geographical determinants of onchocerciasis transmission in a forest/savannah transition zone : two villages of the mbam focus

Mbam, a focus of onchocerciasis outbreaks, is located in the forest/savannah transition zone of Cameroon. Transmission of the disease in this area is similar to that sometimes observed in West Africa. Geographical factors affecting transmission must be identified to determine the areas for priority action. The locations of breeding sites for black flies (the vector of the disease) and factors favorable to their dispersion, the social behavior of these population and the way that she occupy space, must be investigated to identify areas in which the flies and man are in close contact. This geographical study shows that the closest contact between men and black flies occurs in cocoa plantations, due to the proximity of the breeding sites. The areas of closest contact for women and children are unknown, but their identification is important because women and children may be bitten by the flies during their daily activities.

The urgent need for environmental sanitation and a safe drinking water supply in Mbandjock, Cameroon.

Studies were conducted to assess the physical, chemical, and bacteriological qualities of drinking water in Mbandjock, Cameroon. Study results indicated that the vast majority of drinking water sources possessed acceptable physical and chemical qualities, according to the World Health Organization standards. However, microbiological analyses revealed that only the waters treated by the Cameroon National Water Company (SNEC) and the Sugar Processing Company (SOSUCAM) were acceptable for human consumption. All spring and well waters presented evidences of fecal contamination from human and/or animal origin. Water from these sources should, therefore, be treated before use for drinking. Since the majority of the population gets its water from wells and springs, there is an urgent need to develop a health education program, within the framework of primary health care, with respect to environmental sanitation and safe drinking water supply in this community.

International indoor and outdoor evaluation of Bacillus sphaericus products: complexity of standardizing outdoor protocols.

Only one Bacillus sphaericus strain, strain 2362, is currently used commercially to control Culex larval populations. A reliable methodology, easily used, was developed to identify new strains for field application. Larvicidal activities of 3 highly mosquitocidal strains, strains C3-41, Mal, and LB24, previously selected in the laboratory, were compared with that of strain 2362 in tropical and European countries. The following steps were performed: production and titration of acetonic powders from these 4 strains on local Culex species, survey of initial and residual activity under standardized indoor and outdoor conditions, and evaluation of the efficacy of liquid formulations of the 4 strains in natural breeding sites of Culex. In indoor conditions, strain C3-41 showed the highest activity on both Culex pipiens and Culex quinquefasciatus; strain Mal was the least active. The residual activity causing 80% mortality differed from 20 to 90 days according to the strains and the country. Outdoor experiments with powders (0.02-1.6 mg/liter) were performed and the initial toxicities were similar in all cases. Residual activities were very different, from 6 to 95 days posttreatment. Liquid formulations were applied to larval habitats (from 0.1 to 10 g/m2). In tropical countries, larval recolonization in cesspits or ponds occurred after 10-35 days. In Europe, higher doses were needed in polluted water than in clear water (from 3 to 10 liter/ha) for the same control, and the time before 80% residual activity was reached was less than 9-12 days. However, in cesspits, residual activity could be observed for 12 days to 5 mo. A strain 3-5 times more active than the others in bioassays is not significantly detectable from those strains in field trials.

Control of Culex quinquefasciatus (Diptera: Culicidae) with Bacillus sphaericus in Maroua, Cameroon.

Two strategies were tested to control Culex quinquefasciatus with Bacillus sphaericus in Maroua (population 130,000), Cameroon. The treatment of all potential breeding sites (27,000) with B. sphaericus during the dry season caused up to a 90% reduction in the adult biting rate. Because of the short persistence of B. sphaericus and the occurrence of new breeding sites, unacceptable levels of adult biting rates were reached again in 5 months. In the second strategy, two treatments per year of the most productive breeding sites (10,000) stopped the biting rate increase during the rainy season. The results were only partially successful because of variations in B. sphaericus toxicity. The first treatment required 1,200 man-days of work vs. 200 for the simplified treatments. The density of breeding sites depends on the rainfall and the presence of a tap-water network. A sustained control program of Cx. quinquefasciatus will depend upon the dynamics of the principal breeding sites and an improved formulation of B. sphaericus.

Ultra-rapid, simple, sensitive, and economical silica method for extraction of dengue viral RNA from clinical specimens and mosquitoes by reverse transcriptase-polymerase chain reaction.

A rapid, simple and efficient single-tube procedure is described for the isolation of dengue virus RNA from small amount of serum (10 microliters) followed by a reverse transcriptase-polymerase chain reaction (RT-PCR). Recovery of RNA is based on the lysing and nuclease-inactivating properties of guanidinium thiocyanate in the presence of silica. The silica RT-PCR can be completed within 5 hours starting from RNA extraction to agarose gel electrophoresis. All of the 63 dengue-3 culture-positive sera were RT-PCR-positive (virus titres: < 10(2) to 11(10.69.). Of 33 culture-negative acute sera from serologically confirmed dengue fever patients collected during dengue-3 epidemic, 4 were RT-PCR-positive. RT-PCR was also positive in 29 of 30 dengue-1 culture-positive sera (virus titres range: < 10(2) to 10(8.69). Dengue-1 virus was also detected in field-caught Aedes aegypti mosquitoes by silica RT-PCR.

[Campaign against Culex quinquefasciatus using Bacillus sphaericus: results of a pilot project in a large urban area of equatorial Africa]. / Lutte contre Culex quinquefasciatus par Bacillus sphaericus: résultats d'une campagne pilote dans une grande agglomération urbaine d'Afrique équatoriale.

Culex quinquefasciatus, which is sometimes the vector of Bancroft's filariasis, is a harmful mosquito, the immature stages of which live in collections of waste water resulting from human activity. Larval control, the most appropriate method, is at present carried out with chemical insecticides. But the toxicity of these compounds, together with phenomena of resistance, and the cost of substitute insecticides have turned research towards products of biological origin, and one of the most promising is a liquid concentrate of Bacillus sphaericus strain 2362. This was applied experimentally over an area of 200 hectares in a large city in the south of Cameroon, characterized by a short dry season during which mosquito density is at its highest. Spraying was carried out every three months for a year in a concentration of 10 g/m2 and its efficacy evaluated at the level of adult mosquitos through an indirect system of capture on human baits. The results of this study show, in essence, a reduction by 52.7% in the number of females captured in the overall study area treated, and that the impact of treatment is greater during the period of high mosquito density (55.1%) than in the low density period (40.7%). After analysis of the full set of results, the authors conclude that the pattern of rainfall, the conditions in which the insecticide is applied and reinvasions of mosquitos from untreated areas were the principal factors limiting the efficacy of this control campaign.(ABSTRACT TRUNCATED AT 250 WORDS)

Dengue 1 epidemic in French Polynesia, 1988-1989: surveillance and clinical, epidemiological, virological and serological findings in 1752 documented clinical cases.

An epidemic of dengue 1 occurred in French Polynesia in December 1988 and June 1989. This paper records (i) the trend of the outbreak and its surveillance and (ii) the clinical, epidemiological and virological data obtained from 1752 documented cases. The epidemic reached its peak in February in Tahiti Island, 7 weeks after its recognition. Among 6034 suspect cases reported by sentinel physicians, 60.3% were < 20 years old. The illness was classical dengue. No fatality or case of dengue haemorrhagic fever/dengue with shock syndrome was reported. Of 4792 patients subjected to laboratory testing, 41% were confirmed as positive. The serological attack rate was c. 40%. The estimated number of dengue infections in the Windward Islands was about 20,000. Transmission was associated with Aedes aegypti. Study of documented cases showed a higher confirmation rate in both the civilian population < 15 years old (46.5%) and the susceptible French military population (47.6%) than in older civilians (31.1%, P < 0.05). Furthermore, primary dengue infections were predominant in both of the first 2 groups. The diagnosis was mostly confirmed (i) by virus isolation on day < 5 of illness and (ii) by detection of immunoglobulin (Ig) M on day > or = 5 of illness. The study showed that adequate surveillance of an epidemic requires both clinically and laboratory-based systems.

Cumulative mortality rates in Aedes polynesiensis after feeding on polynesian Wuchereria bancrofti carriers treated with single doses of ivermectin, diethylcarbamazine and placebo.

During a therapeutic trial, batches of 672 to 1979 laboratory-bred Aedes polynesiensis, the mosquito vector of lymphatic filariasis in French Polynesia, were fed on Wuchereria bancrofti carriers one, three and six months after they had been treated with either single doses of ivermectin at 100 mcg/kg, diethylcarbamazine (DEC) at 3 and 6 mg/kg or placebo. High mortality rates were observed during the 15-day period following the blood-meal in mosquitoes fed on carriers treated with microfilaricidal drugs and were significantly higher in mosquitoes fed on carriers treated with ivermectin than in those fed on carriers treated with DEC. Though its intensity decreased with the passage of time, the phenomenon was observed in mosquitoes fed on carriers up to six months after treatment, especially in those fed on carriers treated with ivermectin. By decreasing the number of mosquitoes able to transmit the infection, this lethal effect on Ae. polynesiensis might represent an additional advantage of ivermectin in lymphatic filariasis control programs.