Establishing an insect disease vector functional genomics training center in Africa.

The genome sequences for many insects vector of human diseases are now available and promise the development of a set of new, powerful tools that can be used to develop innovative approaches to control these diseases. The African continent, which is the most severely affected by vector borne diseases, lacks adequate infrastructures and personal resources required for rational use of genomic information. To fill this gap, the African Center for Training in Functional Genomics of Insect vectors of Human Disease (AFRO VECTGEN) was initiated by WHO/TDR and the Department of Medical Entomology and Vector Ecology (DMEVE) of the Malaria Research and Training Center (MRTC) in Mali. The aim of the AFRO VECTGEN program is to train young scientists in functional genomics who will ultimately use genome sequence data for research on insect vector of human disease. The program could trigger collaborative research and will benefit from an existing vector biology network in Mali, which was built around research grants funded by the National Institutes of Health, USA and WHO/TDR.

[Evaluation of the diagnostic and therapeutic management of sexually transmissible infections at the Gabriel Touré University Hospital]. / Evaluation de la prise en charge diagnostique et therapeutique des infections sexuellement transmissibles au CHU Gabriel Toure.

PURPOSE: To report cases of sexually transmitted infections (STIs) in consultation and evaluate syndromic management. PATIENTS AND METHODS: We carried out a prospective and descriptive study in patients who had consulted for STI at the urology department of the Gabriel Toure University Hospital from April 2013 to March 2014.HIV cases were not included. The syndromic approach was used. RESULTS: We obtained the following results: 460 cases of STI (Dysuria, Cystitis, Scrotal swelling, Prostatitis). The sex ratio was 20.52 in favor of the male sex. The age group 23-32 years was the most affected, and non-married single patients were the most represented. The majority were Bambara. All cases were diagnosed clinically. Additional tests were requested to confirm the diagnosis. In terms of syndromic management, probabilistic antibiotic therapy was applied to each type of STI. Antibiotic therapy was rehabilitative with an effective therapeutic response in 61.61% of cases for prostatitis, 98.60% for urethritis, 100% for scrotal swelling, 90.78% for cystitis. CONCLUSION: Management of STIs using the syndromic approach is an option in countries where laboratory diagnosis is not always available.

BUT: Il s'agissait de rapporter des cas d'infection sexuellement transmissible (IST) en consultation et d'évaluer la prise en charge syndromique. PATIENTS ET MÉTHODES: Nous avons effectué une étude prospective et descriptive chez les patients qui avaient consulté pour IST au service d'urologie du CHU Gabriel TOURE d'Avril 2013 à Mars 2014.Les cas VIH n'ont pas été inclus. L'approche syndromique a été utilisée. RÉSULTATS: Nous avons obtenu les résultats suivants: 460 cas d'IST (Dysurie, Cystite, Tuméfaction scrotale, Prostatite). Le sexe ratio était de 20,52 en faveur du sexe masculin. La tranche d'âge 23­32 ans était la plus touchée, les patients célibataires étaient les plus représentés. La majorité était bambara. Tous les cas avaient été diagnostiqués cliniquement. Les examens complémentaires étaient demandés pour avoir une confirmation du diagnostic. Sur le plan de la prise en charge syndromique, l'antibiothérapie probabiliste a été appliquée à chaque type IST. L'antibiothérapie, avait été réadaptée à l'antibiogramme avec une réponse thérapeutique efficace dans: 61,61% pour les prostatites, 98,60% pour l'urétrite, 100% tuméfaction scrotale, 90, 78% pour la cystite. CONCLUSION: La prise en charge des IST selon l'approche syndromique est une option dans les pays où le diagnostic de laboratoire n'est pas toujours disponible.

Entomological and parasitological parameters of malaria transmission in Douguia, Chad.

Malaria remains a threat for many countries, especially in Chad where it is the leading cause of morbidity and mortality. Few reliable data exist, however, about the entomological and parasitological parameters of malaria transmission. The objective of this study was to investigate the entomological and parasitological parameters of malaria transmission in Douguia, a village located 75 km northeast of Ndjamena (Chad), as part of a training program for participants in Chad's malaria control program. Mosquitoes were collected after morning pyrethroid spraying, with a mouth aspirator. The parasitological data were collected by a rapid diagnosis test or microscopic examination. The study examined 350 subjects aged from 6 months to 80 years. The plasmodic index (PI) of Plasmodium falciparum was 25.4 % (n = 89) and the gametocygenic index (GI) 9.1 % (n = 32); they varied significantly from one age group to another (p = 10- 3). The PI in pregnant women attending antenatal clinics was 18.7 % (n = 12/64). Three Anopheles species were found: Anopheles gambiae s.l, An. arabiensis and An. pharoensis. An. coluzzii accounted for 94.9 % of the An. gambiae s.s. The antigen sporozoite index (SI) was 4.5 %. Our results confirm the endemicity of malaria in Chad (Douguia) and underline the major role of An. gambiae s.l. in its transmission. However, repeated studies using PCR for Plasmodium detection would help to improve our understanding of its epidemiology.

Gene flow among populations of the malaria vector, Anopheles gambiae, in Mali, West Africa.

The population structure of the Anopheles gambiae complex is unusual, with several sibling species often occupying a single area and, in one of these species, An. gambiae sensu stricto, as many as three "chromosomal forms" occurring together. The chromosomal forms are thought to be intermediate between populations and species, distinguishable by patterns of chromosome gene arrangements. The extent of reproductive isolation among these forms has been debated. To better characterize this structure we measured effective population size, N(e), and migration rates, m, or their product by both direct and indirect means. Gene flow among villages within each chromosomal form was found to be large (N(e)m > 40), was intermediate between chromosomal forms (N(e)m approximately 3-30), and was low between species (N(e)m approximately 0.17-1.3). A recently developed means for distinguishing among certain of the forms using PCR indicated rates of gene flow consistent with those observed using the other genetic markers.

The spatial distribution of Anopheles gambiae sensu stricto and An. arabiensis (Diptera: Culicidae) in Mali.

Variations in the biology and ecology and the high level of genetic polymorphism of malaria vectors in Africa highlight the value of mapping their spatial distribution to enhance successful implementation of integrated vector management. The objective of this study was to collate data on the relative frequencies of Anopheles gambiae s.s. and An. arabiensis mosquitoes in Mali, to assess their association with climate and environmental covariates, and to produce maps of their spatial distribution. Bayesian geostatistical logistic regression models were fitted to identify environmental determinants of the relative frequencies of An. gambiae s.s. and An. arabiensis species and to produce smooth maps of their geographical distribution. The frequency of An. arabiensis was positively associated with the normalized difference vegetation index, the soil water storage index, the maximum temperature and the distance to water bodies. It was negatively associated with the minimum temperature and rainfall. The predicted map suggests that, in West Africa, An. arabiensis is concentrated in the drier savannah areas, while An. gambiae s.s. prefers the southern savannah and land along the rivers, particularly the inner delta of Niger. Because the insecticide knockdown resistance (kdr) gene is reported only in An. gambiae s.s. in Mali, the maps provide valuable information for vector control. They may also be useful for planning future implementation of malaria control by genetically manipulated mosquitoes.

Patterns of irrigated rice growth and malaria vector breeding in Mali using multi-temporal ERS-2 synthetic aperture radar.

We explored the use of the European Remote Sensing Satellite 2 Synthetic Aperture Radar (ERS-2 SAR) to trace the development of rice plants in an irrigated area near Niono, Mali and relate that to the density of anopheline mosquitoes, especially An. gambiae. This is important because such mosquitoes are the major vectors of malaria in sub-Saharan Africa, and their development is often coupled to the cycle of rice development. We collected larval samples, mapped rice fields using GPS and recorded rice growth stages simultaneously with eight ERS-2 SAR acquisitions. We were able to discriminate among rice growth stages using ERS-2 SAR backscatter data, especially among the early stages of rice growth, which produce the largest numbers of larvae. We could also distinguish between basins that produced high and low numbers of anophelines within the stage of peak production. After the peak, larval numbers dropped as rice plants grew taller and thicker, reducing the amount of light reaching the water surface. ERS-2 SAR backscatter increased concomitantly. Our data support the belief that ERS-2 SAR data may be helpful for mapping the spatial patterns of rice growth, distinguishing different agricultural practices, and monitoring the abundance of vectors in nearby villages.

DNA analysis of transferred sperm reveals significant levels of gene flow between molecular forms of Anopheles gambiae.

Anopheles gambiae populations in west Africa are complex, being composed of multiple, sympatric subpopulations. Recent studies have failed to reveal significant genetic differences among subpopulations, stimulating a debate regarding the levels of gene flow among them. The observed homogeneity may be the consequence of substantial contemporary gene flow or it may be that reproductive isolation is complete, but too recent for the accumulation of significant levels of genic divergence. Here, we report the results of a study estimating contemporary levels of gene flow between An. gambiae subpopulations by analysing females and transferred sperm removed from their reproductive systems. A total of 251 female and associated sperm extracts was analysed from a single site in Mali. Two molecular forms of An. gambiae, the M- and S-forms, occurred in sympatry at this site. Overall, we found very strong positive assortative mating within forms, however, we did observe significant hybridization between forms. In the M subpopulation 2/195 females (1.03%) contained sperm from S-form males and in 55 S-form females we found one female containing M-form sperm (1.82%). We also identified a mated M xS hybrid adult female. From mating frequencies, we estimate the Nem between the M- and S-form at 16.8, and from the adult hybrid frequency at 5.6. These values are consistent with our earlier estimate, based on FST for 21 microsatellite loci in which Nem = 5.8. We conclude that the general lack of genetic divergence between the M and S subpopulations of An. gambiae can be explained entirely by contemporary gene flow.

Genetic differentiation of Anopheles gambiae s.s. populations in Mali, West Africa, using microsatellite loci.

Anopheles gambiae sensu stricto is a principal vector of malaria through much of sub-Saharan Africa, where this disease is a major cause of morbidity and mortality in human populations. Accordingly, population sizes and gene flow in this species have received special attention, as these parameters are important in attempts to control malaria by impacting its mosquito vector. Past measures of genetic differentiation have sometimes yielded conflicting results, in some cases suggesting that gene flow is extensive over vast distances (6000 km) and is disrupted only by major geological disturbances and/or barriers. Using microsatellite DNA loci from populations in Mali, West Africa, we measured genetic differentiation over uniform habitats favorable to the species across distances ranging from 62 to 536 km. Gene flow was strongly correlated with distance (r(2) = 0.77), with no major differences among chromosomes. We conclude that in this part of Africa, at least, genetic differentiation for microsatellite DNA loci is consistent with traditional models of isolation by distance.

The pyrethroid knock-down resistance gene in the Anopheles gambiae complex in Mali and further indication of incipient speciation within An. gambiae s.s.

In Mali the Anopheles gambiae complex consists of An. arabiensis and Mopti, Savanna and Bamako chromosomal forms of An. gambiae s.s. Previous chromosomal data suggests a complete reproductive isolation among these forms. Sequence analysis of rDNA regions led to the characterization of two molecular forms of An. gambiae, named M-form and S-form, which in Mali correspond to Mopti and to Savanna/Bamako, respectively, while it has failed so far to show any molecular difference between Savanna and Bamako. The population structure of An. gambiae s.l. was analysed in three villages in the Bamako and Sikasso areas of Mali and the frequency of pyrethroid resistance of the knock-down resistance (kdr) type was calculated. The results show that the kdr allele is associated only with the Savanna form populations and absent in sympatric and synchronous populations of Bamako, Mopti and An. arabiensis. This is the first molecular indication of barriers to gene flow between the Bamako and Savanna chromosomal forms. Moreover, analyses of specimens collected in the Bamako area in 1987 show that the kdr allele was already present in the Savanna population at that time, and that the frequency of this allele has gradually increased since then.

Complexities in the genetic structure of Anopheles gambiae populations in west Africa as revealed by microsatellite DNA analysis.

Chromosomal forms of Anopheles gambiae, given the informal designations Bamako, Mopti, and Savannah, have been recognized by the presence or absence of four paracentric inversions on chromosome 2. Studies of karyotype frequencies at sites where the forms occur in sympatry have led to the suggestion that these forms represent species. We conducted a study of the genetic structure of populations of An. gambiae from two villages in Mali, west Africa. Populations at each site were composed of the Bamako and Mopti forms and the sibling species, Anopheles arabiensis. Karyotypes were determined for each individual mosquito and genotypes at 21 microsatellite loci determined. A number of the microsatellites have been physically mapped to polytene chromosomes, making it possible to select loci based on their position relative to the inversions used to define forms. We found that the chromosomal forms differ at all loci on chromosome 2, but there were few differences for loci on other chromosomes. Geographic variation was small. Gene flow appears to vary among different regions within the genome, being lowest on chromosome 2, probably due to hitchhiking with the inversions. We conclude that the majority of observed genetic divergence between chromosomal forms can be explained by forces that need not involve reproductive isolation, although reproductive isolation is not ruled out. We found low levels of gene flow between the sibling species Anopheles gambiae and Anopheles arabiensis, similar to estimates based on observed frequencies of hybrid karyotypes in natural populations.

Mapping rice field anopheline breeding habitats in Mali, West Africa, using Landsat ETM+ sensor data.

The aim of this study was to determine whether remotely sensed data could be used to identify rice-related malaria vector breeding habitats in an irrigated rice growing area near Niono, Mali. Early stages of rice growth show peak larval production, but Landsat sensor data are often obstructed by clouds during the early part of the cropping cycle (rainy season). In this study, we examined whether a classification based on two Landsat Enhanced Thematic Mapper (ETM)+ scenes acquired in the middle of the season and at harvesting times could be used to map different land uses and rice planted at different times (cohorts), and to infer which rice growth stages were present earlier in the season. We performed a maximum likelihood supervised classification and evaluated the robustness of the classifications with the transformed divergence separability index, the kappa coefficient and confusion matrices. Rice was distinguished from other land uses with 98% accuracy and rice cohorts were discriminated with 84% accuracy (three classes) or 94% (two classes). Our study showed that optical remote sensing can reliably identify potential malaria mosquito breeding habitats from space. In the future, these 'crop landscape maps' could be used to investigate the relationship between cultivation practices and malaria transmission.

Mark-release-recapture experiments with Anopheles gambiae s.l. in Banambani Village, Mali, to determine population size and structure.

Mark-release-recapture experiments with Anopheles gambiae s.l. were performed during the wet seasons of 1993 and 1994 in Banambani, Mali. All recaptured mosquitoes were identified to species by PCR analysis and, when possible, by chromosomal analysis to chromosomal form. Two species of the An. gambiae complex were present: An. gambiae s.s. and An. arabiensis; their ratio differed greatly from one year to the next. Three chromosomal forms of An. gambiae s.s. were found--Bamako, Savanna and Mopti. The drier 1993 was characterized by a high frequency of An. arabiensis and of the Mopti chromosomal forms of An. gambiae s.s. These trends were consistent with large-scale geographical patterns of abundance along a precipitation gradient. We observed no difference in dispersal between the two species, nor among the chromosomal forms of An. gambiae s.s. Therefore, in this situation at least, it is reasonable to group such data on the An. gambiae complex as a whole for analysis. Population size of An. gambiae s.l. females in the village was estimated to be 9000-11,000 in 1993 and 28,000 in 1994. The corresponding numbers were somewhat higher when independently-derived values of daily survival were used. These were consistent with estimates of effective population size obtained from patterns of gene frequency change.