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Abderrazak, S. B., Oury, B, Lal, A. A., Bosseno, M.-F., Force-Barge, P., Dujardin, J.-P., Fandeur, T., Molez, J.-F., Kjellberg, F., Ayala, F. J., and Tibayrenc, M. 1999. Plasmodium falciparum: Population genetic analysis by multilocus enzyme electrophoresis and other molecular markers. Experimental Parasitology 92, 232-238. The population structure of Plasmodium falciparum, the agent of malignant malaria, is uncertain. We have analyzed multilocus enzyme electrophoresis (MLEE) polymorphisms at 7-12 gene loci in each of four populations (two populations in Burkina Faso, one in Sudan, one in Congo), plus one "cosmopolitan" sample consisting of parasite cultures from 15 distant localities in four different continents. We have also performed random amplified polymorphic DNA analysis (RAPD) and restriction fragment length polymorphism (RFLP) and characterized gene varia tion at four antigen genes in the Congo population. All genetic assays show abundant genetic variability in all populations analyzed. With the isoenzyme assays, strong linkage disequilibrium is apparent in at least two local populations, the Congo population and one population from Burkina Faso, as well as in the cosmopolitan sample, and less definitely in the other Burkina Faso population. However, no linkage disequilibrium is detected in the Congo population with the molecular assays. We failed to detect any nonrandom association between the different kinds of genetic markers; that is, MLEE with RAPD or RFLP, RAPD with RFLP, and so on. Although isoenzyme data show statistical departures from panmictic expectations, these results suggest that in the areas under survey, P. falciparum populations do not undergo predominant clonal evolution and show no clear-cut subdivisions, un like Trypanosoma cruzi, Leishmania sp., and other major parasitic species. We discuss the epidemiological and taxonomical significance of these results.

Background: It is recommended that children aged 3 months to five years of age living in areas of seasonal transmission in the sub-Sahel should receive Seasonal Malaria Chemoprevention (SMC) with sulfadoxine-pyrimethamine plus amodiaquine (SPAQ) during the malaria transmission season. The purpose of this study was to evaluate the safety of SMC with SPAQ in children when delivered by community health workers in three districts in Senegal where SMC was introduced over three years, in children from 3 months of age to five years of age in the first year, then in children up to 10 years of age., Methods: A surveillance system was established to record all deaths and all malaria cases diagnosed at health facilities and a pharmacovigilance system was established to detect adverse drug reactions. Health posts were randomized to introduce SMC in a stepped wedge design. SMC with SPAQ was administered once per month from September to November, by nine health-posts in 2008, by 27 in 2009 and by 45 in 2010., Results: After three years, 780,000 documented courses of SMC had been administered. High coverage was achieved. No serious adverse events attributable to the intervention were detected, despite a high level of surveillance., Conclusions: SMC is being implemented in countries of the sub-Sahel for children under 5 years of age, but in some areas the age distribution of cases of malaria may justify extending this age limit, as has been done in Senegal. Our results show that SMC is well tolerated in children under five and in older children. However, pharmacovigilance should be maintained where SMC is implemented and provision for strengthening national pharmacovigilance systems should be included in plans for SMC implementation., Trial Registration: ClinicalTrials.gov NCT 00712374., Competing Interests: The authors have declared that no competing interests exist.

Purpose: The authors studied the role of bacteria belonging to Anaplasmataceae family as the causes of acute illnesses of sheep in West Africa., Methods: We examined and sampled 120 febrile sheep in two regions of Senegal for this study. The DNA extracted from these blood samples was tested by PCR using two pairs of primers (groEL-based and 16S rRNA gene-based)., Results: In 52/120 samples, the microscopic examination revealed intraerythrocytic and/or intraphagocytic spherical inclusions. In 48/52 cases, we succeeded in identifying the bacterial agent: in 38 cases, it was Anaplasma ovis; in six cases, it was Ehrlichia ruminantium; in two cases, Anaplasma phagocytophilum; in one case, Anaplasma platys; and in one case, a yet uncultured Anaplasma sp. closely related to A. phagocytophilum., Conclusions: Our studies demonstrated the great variety of pathogenic bacteria from the Anaplasmataceae family in the blood of clinically ill sheep. A. ovis was identified unexpectedly often. For the first time, A. phagocytophilum was found in sub-Saharan Africa, and its further epidemiology may be now reconsidered. The roles of canine pathogen, A. platys, and yet undescribed Anaplasma sp. "Badiouré" in ovine pathology should be more closely studied.

Background: The Demographic Surveillance System established in 1962 in Niakhar, Senegal, is the oldest in Africa. Here, we analyze trends in overall child mortality, malaria, and other causes of death in Niakhar from the beginning of data collection to 2010., Methods: After an initial census, demographic data were updated yearly from 1963 through 2010. From 1984, causes of death were determined by the verbal autopsy technique., Results: During 1963-2010, infant and under-5 mortality rates decreased from 223‰ to 18‰ and from 485‰ to 41‰, respectively. The decrease was progressive during the entire observation period, except during 1990-2000, when a plateau and then an increase was observed. Malaria-attributable mortality in under-5 children decreased from 13.5‰ deaths per 1000 children per year during 1992-1999 to 2.2‰ deaths per 1000 children per year in 2010. During this period, all-cause mortality among children aged <5 years decreased by 80%., Conclusions: Inadequate treatment for chloroquine-resistant malaria and an epidemic of meningitis during the 1990s were the 2 factors that interrupted a continuous decrease in child mortality. Direct and indirect effects of new malaria-control policies, introduced in 2003 and completed during 2006-2008, are likely to have been the key cause of the recent dramatic decrease in child mortality.

In the context of global warming and the risk of spreading arthropod-borne diseases, the emergence and reemergence of leishmaniasis should not be neglected. In Senegal, over the past few years, cases of canine leishmaniasis have been observed. We aim to improve the understanding of the transmission cycle of this zoonosis, to determine the responsible species and to evaluate the risk for human health. An epidemiological and serological study on canine and human populations in the community of Mont Rolland (Thiès area) was conducted. The data showed a high seroprevalence of canine leishmaniasis (>40%) and more than 30% seropositive people. The dogs' seroprevalence was confirmed by PCR data (concordance > 0.85, Kappa > 0.7). The statistical analysis showed strong statistical associations between the health status of dogs and seropositivity, the number of positive PCRs, clinical signs and the number of Leishmania isolates. For the first time, the discriminative PCRs performed on canine Leishmania strains clearly evidenced that the pathogenic agent is Leishmania infantum. The results obtained show that transmission of this species is well established in this area. That the high incidence of seropositivity in humans may be a consequence of infection with this species is discussed., (Copyright © 2010 Institut Pasteur. Published by Elsevier SAS. All rights reserved.)

Background: Q fever is a worldwide zoonotic disease caused by Coxiella burnetii. Epidemiologically, animals are considered reservoirs and humans incidental hosts., Methodology/principal Findings: We investigated Q fever in rural Senegal. Human samples (e.g., sera, saliva, breast milk, feces) were screened in the generally healthy population of two villages of the Sine-Saloum region. Ticks were collected in four regions. Seroprevalence was studied by immunofluorescence, and all other samples were tested by two qPCR systems for detection of C. burnetii. Positive samples were genotyped (multispacer typing) by amplification and sequencing of three spacers. Strains were isolated by cell culture. We found that the seroprevalence may be as high as 24.5% (59 of 238 studied) in Dielmo village. We identified spontaneous excretion of C. burnetii by humans through faeces and milk. Hard and soft ticks (8 species) were infected in 0-37.6%. We identified three genotypes of C. burnetii. The previously identified genotype 6 was the most common in ticks in all studied regions and the only one found in human samples. Three strains of genotype 6 of C. burnetii were also recovered from soft tick Ornithodoros sonrai. Two other genotypes found in ticks, 35 and 36, were identified for the first time., Conclusions/significance: Q fever should be considered a significant public health threat in Senegal. Humans, similar to other mammals, may continuously excrete C. burnetii.

By using degenerate primers deduced from conserved patterns in the flavivirus polymerase gene, a novel RNA virus was discovered in Rhipicephalus ticks sampled from members of the family Bovidae in Senegal. It was named Ngoye virus (NGOV) after the location from which it was isolated. Viral particles could be observed by electron microscopy, but isolation in vertebrate or invertebrate cell lines or by intracerebral infection of newborn mice remained unsuccessful. This is atypical of recognized arboviruses. The characterization of 4176 nt of the non-structural genes revealed that NGOV is a novel flavivirus species. It forms a distinct phylogenetic lineage related distantly to previously identified members of the genus Flavivirus. Analysis of genetic data suggested that the processing of the NGOV polyprotein and the organization of its replication complex are similar to those of flaviviruses. Together with other recent data, these findings suggest that a large number of viruses related distantly to 'classical' arthropod-borne flaviviruses remain to be discovered.

A historical and comparative study of the origins and emergence of syphilis and AIDS show that both result from human intrusions. Treponema probably existed in primates before human infection, and nonvenereal treponemal infection existed in prehistoric tropical Africa. When humans began wearing clothes, the disappearance of endemic infection ended immunity and led to receptivity to venereal infection. It was long thought that syphilis was first introduced in Europe by the conquistadors, but lesions typical of treponematosis dating from before the Common Era have been found in Europe. It is possible that the first navigators transferred treponemal infections to Latin America. AIDS seems to have appeared throughout the Congo River basin around 1950, and genetic studies attest to its long history in primates. It may have resulted from the Bantu migration and its strong human intrusion into the forest. After the initial human infection, new epidemiological factors in a transformed environment and behavioral changes led at 500-year intervals (1480-1490 for syphilis and 1940-1950 for AIDS) to the widespread emergence and subsequent pandemic of each disease.

This study of malaria biodiversity in Senegal used an entomological approach that combined parasite surveys and clinical investigations in the mangrove area of the Saloum delta from 1996 to 1998. The parasitologic studies took place in two of the six villages in the coastal area of Palmarin (Djifère and Diakhanor) during three distinct periods: at the end of the dry season, in the middle of the rainy season, and at the end of the rainy season. The clinical investigations at the Palmarin health station took place from July 1996 through February 1998. A malaria attack was defined as the presence of malaria symptoms (including fever, headaches, sweating, and shivering) associated with plasmodic parasitemia > 3,000 trophozoites/microL of blood. All the positive thick smears were infected with Plasmodium falciparum, one also with P. falciparum, and none with P. ovale. The average plasmodic index (5.6%) classifies the delta of Saloum as a hypoendemic area. The average parasite load was estimated at 2,239 trophozoites (95% CI: 1,660-3,020) of P. falciparum per microliter of blood, and 86.9% of patients with symptoms of a malaria attack were febrile. Malaria attacks accounted for 1.9% of the total consultations, 12.2% of the presumed malaria cases, and 14.0% of the febrile subjects. The finding that malaria attacks affected all age groups confirms the weakness of anti-malaria immunity among the population of the Saloum delta. Malaria cases were more frequent at the end of the rainy season and the beginning of the dry season, periods when parasite loads were highest. In this area, which is increasingly attractive to tourists and has a quite superficial fresh water table, man-made environmental changes favor mosquito breeding sites that promote the development of An. arabiensis and An. gambiae spp, both known to be major malaria vectors. In view of the population's weak anti-malaria immunity, this situation may increase malaria transmission and could be followed by epidemics. It is therefore important to set up a functional system of epidemiological monitoring to detect any malaria outbreaks.

A study of malaria morbidity was carried out from November 1994 to October 1995, in a Ferlo village (Barkedji) characterized by a long persistence of the temporary ponds. The objective was to evaluate the repercussions of the strong and long anopheles transmission in humans. A clinical follow-up of a group of residents was conducted at home every 10 days by an investigator trained for taking axillary temperature and making thick smears, when suspecting malaria. Were included in the group, 123 voluntary subjects among whom 50% were children under 10 years old. Any feverish subject (T degree >37 degrees 5) or subject presenting other malaria symptoms (headaches, hot body shivers, sweats, aches...) was regarded as having a malaria attack as well as a parasitemia >2500 P/mm3 in children aged of 0 to 14 years old and 1000 P/mm3 in the oldest. During the study subjects with at least one feverish access, plasmodium infection and malaria attack were 58%, 33% and 22%, respectively. On 172 hyperthermias observed, 49% were accompanied by a circulating parasitemia and 30% corresponded to malaria attack. The feverish subjects (74% vs. 42%), the subjects with parasitemia (51% vs. 16%) and the cases of malaria (34% vs. 10%) were more frequently encountered in children under10 than in the oldest. The cases of malaria attacks were more frequent from November to January (70%). The strong intensity of malaria transmission in Barkedji and the persistence of its temporary ponds until January were sufficient to influence the level of malaria morbidity and consequently the development of an anti-malaria immunity by the indigenous population.

During the first four months of 2003, the survey laboratory of the Federal District (LACEN Laboratory of Virology), Brasília, Brazil, isolated ten strains of dengue virus serotype 3, five of them autochthonous, and the remaining ones from cases imported from Tocantins, Goias and Bahia States. The virus isolations were performed in C6/36 cell culture inoculated with total blood collected between the 1st and the 5th days after the onset of the symptoms. The age of the patients varied from 26 to 59 years old. The strains were typed as DEN-3 by indirect immunofluorescence assay using serotype-specific monoclonal antibodies. Viral RNAs were extracted from total blood using the trizol method. The nested RT-PCR method detected DNA products of 290 bp, confirming the serotype identifications. The introduction of DEN-3 in Brazil and especially in the Federal District represents a serious threat, since most people are susceptible to this serotype and many have already been infected by serotypes DEN-1 or DEN-2, thus increasing the risk of epidemic of more severe forms of the disease. The use of a fast and reliable method for continuous monitoring of the circulation of this serotype is of primary importance for the prevention and control of future epidemics.

From June 1995 to January 1998, entomological studies carried out in five villages located in the Delta's Saloum have allowed to better understand the contribution of An. melas Theobald (1903) to malaria transmission in mangrove swamp. Among the five villages studied, three of them (Simal, Djilor and Marlothie) located along the Saloum river, are colonised by An. arabiensis; the two others (Djifere and Diakhanor) located between the sea and the river, are colonised by An. melas. During the rainy season and at the beginning of the dry season, An. melas and An. arabiensis are sympatric. The ratio of An. melas/An. arabiensis increases when we go closer the coast where An. melas becomes quite exclusive. When An. melas is predominant, endophagy, endophily and anthropophily are very marked. The parturity rates are lower in An. melas than in An. arabiensis. In the predominance area of each species, transmission is on the same level. During the period of sympatry, An. arabiensis is responsible for the transmission and when it is absent, An. melas carries on. Transmission occurs from July to March with a maximum at the beginning of the dry season. In the villages of the mangrove swamp, its prolongation until the middle of the dry season is due to An. melas.

To date, no consensus exists on the type of crude Plasmodium falciparum Ags to be used in a standard assay for the evaluation of the overall anti-blood-stage immune response in humans. Comparison of the dose-dependent reactivity of using a pool of hyper-immune Senegalese sera to saponin and water schizont extracts of the Senegalese 07/03 isolate indicated similar reactivity on both types of antigen preparations. Water schizont extracts from three different strains of P. falciparum adapted to in vitro culture probed with a panel of specific mouse antisera and monoclonal antibodies reacting with conserved antigens showed similar antigenic content. Seroreactivity of immune individuals living in three different areas of endemicity was assessed in parallel on water crude extracts. The individual IgG, IgM and IgG subclass antibody responses to the various schizont preparations correlated positively. The specific IgM response was higher on the Senegalese schizont extract than on the FCR3 extract and was highest in Dielmo villagers. The IgG response was similar in all three locations and was strain independent. These results indicate that monitoring IgG antibody levels to the widely distributed FCR3 strain using an easily prepared crude lysate might represent a valuable reference ELISA allowing homogenisation and comparison of data from different laboratories.

Blood-engorged females of An. gambiae s.l., An. funestus, An. pharoensis and An. rufipes caught resting indoors were tested (precipitin or enzyme-linked immunosorbent assay) to determine the source of bloodmeal. The species of the An. gambiae complex fed mainly on human hosts in all prospected areas, except in those of the mid Senegal river valley where an important zoophily was observed within and near the irrigation zone. Among 4,597 blood-engorged females of An. gambiae s.l., 29% fed on cattle with 7.5% of mixed boodmeals, mainly human-bovine or human-equine. The presence of cattle, culicid population densities and individual mosquito safety devices were the most determinant factors of animal deviation. Blood of most domestic animals was found in the stomach of collected females, but according to areas, bovines and equines were the main hosts for zoophilic females of An. gambiae s.l. Females of An. funestus collected in the middle-west were more anthropophilic than those collected in south-eastern areas. An. pharoensis was most prevalent in the Senegal river delta where it was found to be very anthropophilic. An. rufipes was strongly endophilic but exclusively zoophilic in all prospected areas.

Among the Indians Desana's (Tukano amerindians) in the Upper Rio Negro, the interseasonal variation of the malarial fevers were associated with two myths (localised in two distinguishable places). One myth associates the malarial with the rivers which contain "malaria pots". Conception based on an observation of localised water collection in the banks and the rocky rapids ("banks and rocky's fever"). The transmission and the anophelian density present variation between the seasons in relation to the river's level. Another myth associates malarial fevers in the forest, with the song of a frog ("fever's frog") and the flowering and fructification of a tree (Poaqueira sericea Thul.). There is in South America a particular type of forest malaria, known as "Bromelia malaria" and denounced in human and/or simian transmission. This forest malaria is transmitted by the a sub-genus anopheles (Kerteszia) which larval breeding are areal in the canopy. The breeding places are found in the forest in the epiphyte bromeliads. To understand this type of transmission, we must take reference to the previous endomological data at the Upper Oyapock Wayâpi (Tupi amerindians). This Bromelia malaria could fluctuated according larval density variation, related to washing of epiphytes (end of the rainy season) or to their flowering (end of the dry season). The "fever's frog" myth collected at the Desana's in the Upper Rio Negro can be related to the existence of Bromelia malaria in this amazonian habitat. These myths showed the perfect adaptation of the amerindians to their environment and their complete knowledge of the neotropical forest.

This study reports on T-cell proliferative responses to the 19-kDa C-terminal domain of the Plasmodium falciparum merozoite surface protein (MSP1(19)). Three different recombinant proteins were used: an Escherichia coli product expressing the first EGF-like domain and Saccharomyces cerevisiae and baculovirus/insect-cell-produced proteins containing both EGF-like domains, the latter protein being produced with or without N-glycosylation. Cell donors were P. falciparum-immune adults with no recent history of clinical malaria and recruited from three Senegalese settings with different epidemiological parasite transmission. Each mononuclear-blood-cell preparation was stimulated with a range of concentrations of the three proteins. Most subjects' mononuclear cells were reactive to at least one protein, but significant differences in lymphoproliferation were seen between the settings and within individual cultures depending on the protein source and concentration. Importantly, lymphoproliferation indices correlated inversely with the intensity of P. falciparum malaria transmission. When purified T lymphocytes were cultured in the presence of MSP1(19) plus autologous monocytes, B lymphocytes or a proposed CD1+ dendritic-cell population as costimulatory cells, significant differences were observed depending on the individual's previous exposure to parasites. This study shows that the stimulation of lymphocyte proliferation in vitro with MSP1(19) depends on several factors, including epidemiological conditions and protein preparations.

The Sahel region has been suffering from severe drought for the last thirty years, with large deficits and a high degree of variability in the amount of annual rainfall. Agricultural production in the Sahelian zone of the Senegal River valley depends on the flooding of the river. The management of this flooding affects malaria transmission. The area is prone to malaria epidemics because the immunity level of the population is low. We studied epidemiological, meteorological and river level data to identify epidemic risk factors. We propose an epidemiological and managerial system for the early detection of risks and early intervention. This system is based mainly on the water level of the Senegal River and the early detection of unusual increases in the number of cases.

Over 12 years, from 1984 to 1995, we conducted a prospective study of overall and malaria specific mortality among three rural populations in the Sahel, savanna and forest areas of Senegal. The emergence of chloroquine resistance has been associated with a dramatic increase in malaria mortality in each of the studied populations. After the emergence of chloroquine resistance, the risk of malaria death among children 0-9 years old in the three populations was multiplied by 2.1, 2.5 and 5.5, respectively. This is the first study to document malaria mortality at the community level in Africa before and after the emergence of chloroquine resistance. Findings suggest that the spread of chloroquine resistance has had a dramatic impact on the level of malaria mortality in most epidemiological contexts in tropical Africa.

In Europe before the advent of the acquired immunodeficiency syndrome (AIDS), fatal cases of infection with Cryptococcus neoformans resembling acute meningitis were rarely described and never in young adults. However, rapidly fatal cryptococcal meningitis in young Africans has been known to exist in central Africa for at least 30 years, mainly in the lower area of the Congo River basin. Cases have been reported in this area since 1953, particularly in young patients during the 1950s. It is also known that central African AIDS patients frequently suffer from cryptococcosis, and there is a possibility that earlier clinical reports of encephalitis were actually fatal cases of AIDS in young Africans. It appears possible that the central part of the African continent is the area where human immunodeficiency virus originated.

Hispaniola has the highest malaria rate in the West Indies. A study of the bio-ecology of the major vector Anopheles albimanus was carried out in Haïti. Our observations concerned the length of pre-imaginal stages, biting rates and the gonotrophic cycle. A table of its complete life cycle, stage by stage, was produced as well as a graph illustrating its whole larval development in its natural habitat. Results in the field and insectarium were compared. In the laboratory, the development was shorter, except for pupae. The temperature was the main factor governing the length of pre-imaginal development. Preimaginal development decreased from 15 days in the field to 9 days in the insectarium whereas the temperature was a high of 4 degrees C. The nycthemeral activity and seasonal variations of An. albimanus were studied, by monthly all-night captures on human volunteers, during a full year in exo- and endophilic situations. Two sampling sites were chosen for comparison, An. albimanus were caught at every hour of the nycthemeral cycle, with a peak around midnight. Monthly variations were affected by the rains. The vector was more exophagous, when its density was high. The parity rate of the biting population varied between 85.8% and 20.0%; it was low at the beginning of the rains and increased when the rains stopped and breeding places dried out. To study the gonotrophic cycle of wild An. albimanus, several techniques were used jointly; follicles dissection; follicular relics; length of maturation of follicles. The length of the full cycle and of each of its components was estimated (quest of host for blood meal; maturation phase; breeding site seeking). In Haïti 41 h (nulliparous), or 21 h (parous) were necessary for females to have a complete maturation of the follicles. The lengthening of the gonotrophic cycle of parous An. albimanus, in the natural field site, was estimated to last about 4 days. For nulliparous it was 5 days, and 7 days if a pregravid stage was necessary. Considering the sporogonic cycle of Plasmodium falciparum (12 days), the wild population of An. albimanus females could become infective at the third blood feeding in Haïti.

The efficacy of permethrin-treated bednets was evaluated in Wassadou, a hyperendemic village located in the Sudanese grasslands of southeast Senegal. Pretreatment data were collected between 1992 and 1993. Bednets were distributed to the whole population in June 1995 and impact of their use on vector populations and malaria transmission was evaluated until November 1995. This period corresponds to the rainy season during which malaria transmission is highest. Data were compared with a control village in which bednets were not distributed. Findings showed that use of bednets led to a sharp decrease in the density of the vector population and malaria transmission. The number of bites by Anopheles gambiae s.1. decreased 69%. The density of blood-laden and pregnant females inside dwellings decreased 91% and 96% respectively. The sporozoite index of females captured on the skin decreased 76% and the daily rate of entomological inoculation decreased 88%. This impact was not great enough to eliminate the risk of infection. Prolonged study over a period of 4 to 5 years is needed to evaluate the impact of long-term use of insecticide-treated bednets on vector population and malaria transmission.

For the first time in Dakar (Senegal), we managed to establish a local colony of Anopheles arabiensis (Diptera: Culicidae) in insectarium. The observations on the productivity of the colony showed a gradual adaptation of the preimaginal stages to insectarium conditions. The estimated duration of the development of the various preimaginal stages showed an interval of 24 hours for egg hatching, 48 h for stage I, 24 h for stage II, 24 h for stage III and 36 h for the pupal stage. Stage IV which makes the difference in the duration of the adult-egg cycle, varied from 1 to 13 days. Pupal moulting occurred mainly between 11 h and 19 h, with a peak between 14 h and 15 h. The duration of the preimaginal cycle in insectarium was from 8 to 11 days. This fits with the duration shown in natural breeding places during the rainy season (the best period for the development) which was from 9.5 to 13.3 days. Emergence, faster in males, occurred in the evening around 6.30-7 p.m. The emergence rate was higher in insectarium (92.9%) compared with that shown in the field (6 to 20%) (the natural breeding places are subject to biological and biochemical variations and to predators's action). The sex-ratio is equal to 1. The studies in insectarium showed that the gonotrophic cycle is 3 days in neonate blood fed after day 2 and also in parous ones. This cycle is 5 days in nulliparous with pregravid stage. The average longevity is 16 days with a maximum of 26 days in females and 43 days in males.

The ecology, population dynamics, and malaria vector efficiency of Anopheles gambiae and An. arabiensis were studied for 2 yr in a Sahelian village of Senegal. Anophelines were captured at human bait and resting indoors by pyrethrum spray. Mosquitoes belonging to the An. gambiae complex were identified by polymerase chain reaction. Of 26,973 females, An. arabiensis represented 79% of the mosquitoes captured and remained in the study area longer than An. gambiae after the rains terminated. There were no differences in nocturnal biting cycles or endophagous rates between An. gambiae and An. arabiensis. Based on an enzyme-linked immunosorbent assay test of bloodmeals, the anthropophilic rate of these 2 vectors were both approximately 60%, when comparisons were made during the same period. Overall, 18% of the resting females had patent mixed bloodmeals, mainly human-bovine. The parity rates of An. gambiae and An. arabiensis varied temporally. Despite similar behavior, the Plasmodium falciparum circumsporozoite protein (CSP) rates were different between An. gambiae (4.1%) and An. arabiensis (1.3%). P. malariae and P. ovale only represented 4% of the total Plasmodium identified in mosquitoes. Transmission was seasonal, occurring mainly during 4 mo. The CSP entomological inoculation rates were 128 bites per human per year for the 1st yr and 100 for the 2nd yr. Because of the combination of a high human biting rate and a low CSP rate, An. arabiensis accounted for 63% of transmission. Possible origin of differences in CSP rate between An. gambiae and An. arabiensis is discussed in relation to the parity rate, blood feeding frequency, and the hypothesis of genetic factors.

Specific immune responses to asexual blood stages of P. falciparum antigens (a lysate of parasitized red blood cells and a characterized vaccine candidate i.e. MSP1 p19) were analyzed in plasma samples from immune adult individuals living in three different areas of Senegal, where malaria transmission is different. Most individuals in the three sites had specific IgG and IgM to total P. falciparum antigens, whereas approximately 50% had either IgG or IgM specific to MSP1 p19. Further, no anti-MSP1 p19 IgG2 and IgG4 antibody was noticed in any individual whereas the distribution of anti-MSP1 p19 IgG1 and IgG3 was different upon the epidemiological context. In addition, no relationship was found between antibody responses and in vitro T cell responses against P. falciparum antigens upon those experimental conditions. These data stress on the relatively elevated distribution of specific antibodies to MSP1 p19 in P. falciparum hyperendemic areas and suggest a differential regulation of isotypes depending on individual parasite exposure.

The efficacity of oral chloroquine was assessed in 360 out-clinic patients with symptomatic Plasmodium falciparum malaria who were enrolled at five sites, in four administrative regions of Senegal, between 1991 and 1995. They were three rural areas: Mlomp (Casamance), Bandafassilbel (Eastern Senegal), Diohine (Sine-Saloum) and one urban area: Pikine (agglomeration of Dakar). Parasitological failure at Day-7 was observed in 108 patients (30%) and ranged from 14% to 50% according to the study areas. The proportion of RI, RII and RIII responses were 6%, 23% and 1%, respectively. Chloroquine resistance was lowest in Bandafassi, one of the most remote area of Senegal. It was highest in Mlomp where a malaria control programme with mass chemoprophylaxis had been carried out since 1975. The therapeutic failure rate defined by the persistence or reappearance of fever and P. falciparum trophozoites on days 4-7 was 6%. The percentages of therapeutic failure for RI, RII and RIII patients were respectively 14%, 19% and 100%. These results and those of previous studies in Senegal suggest that chloroquine resistance, which first emerged in vivo in 1988 in Dakar, spread between 1990 and 1993 in all regions of this country. The limitations of in vivo tests for the determination of the therapeutic efficacy of chloroquine in malaria endemic regions with increasing chemoresistance are discussed. The low proportion of potentially severe malaria infections in semi-immune persons, the biases in patients selection, and the possibility of delayed complications, considerably limit the potential of these tests for guiding the choice of the best adapted first line treatment.

Patients with Plasmodium falciparum infections were selected with an in vivo chloroquine sensitivity assay. Fourteen days after treatment, the gametocytes were studied in relation to asexual parasite responses classified as drug-sensitive or showing RI or RII resistance. Gametocyte prevalence and density appeared significantly higher in RII than RI strains and higher in RI than in sensitive strains. This finding on gametocyte variation in vivo may explain why the RII type of chloroquine resistance has became more prevalent than RI everywhere in tropical Africa in the short time since its emergence. The biological and epidemiologic advantage of chloroquine-resistant malaria mediated through gametocytes is discussed in the context of the present drug pressure in Africa.

The Niayes is geographically characterized by an alternating succession of sand dunes and wet depressions, 20 km wide, which are behind the offshore bars from Saint-Louis to Dakar. Since 1970, the area has been affected by drought. The rainfall which was around 700 mm per year before 1960 has fallen below 500 mm during the last 30 years. In 1991 it was only 350 mm and in 1992, 260 mm. The vegetation has become impoverished in the depressions which are now more and more cultivated for vegetables. Entomological, parasitological and clinical studies on malaria were carried out in 1991 and 1992 in three villages and the results were compared with data gathered from 1967 to 1968 with comparable methods. The main points concerning the evolution of malaria are listed as follows. An. funestus, which was previously the predominant malaria vector, has almost disappeared. An. gambiae has regressed ahead of its sibling species An. arabiensis, a less competent vector. These changes in vector populations have led to lower malaria transmission and consequently a decrease of the endemicity. The children parasitic index, which was between 40 to 80% according to the localities in 1967, fell to 10% or less in 1991 and 1992. Finally, the incidence of clinical cases, calculated on a cohort of 100 children was only 4% in 1992, which is very low for an African endemic country. We conclude that drought and human activities have modified the environment such that malaria endemicity has seriously decreased. However, the risk of epidemic remains with the eventual occurrence of heavy rains. Therefore a surveillance of the area is needed to prevent a future epidemic.

An epidemiological survey of malaria was carried out from September 1992 to November 1994 in three villages located in the Senegal river delta, two villages growing rice in irrigated fields and one practicing traditional rain water agriculture. Entomological observations showed that Anopheles pharoensis is the main anopheline species caught in the area with a high population density in the rice growing villages. The population density of species of the An. gambiae complex, represented by An. gambiae s.s., An. arabiensis and An. melas is low. Agressivity rates and anthropophlic indexes of An. pharoensis females are high but their parity rates are low. The malaria transmission is weak and was not perceptible in the area as shown by the negative results obtained with the ELISA tests and the examination of salivary glands. Parasitological indexes, malaria morbidity and incidence rates are low and are in agreement with the entomological data. In the Senegal river delta, irrigation has, on the whole, increased the An. pharoensis density but both malaria transmission and incidence rates did not rise.

Some informations about malaria transmission, which has until nox difficult to get, can be obtained thanks to the use of molecular biology tools, PCR mainly. In Senegal, we use that technique to solve two kinds of problems: -Identification of species of the Anopheles gambiae complex: PCR technique is useful compared to other diagnostic methods (chromosome pattern, DNA probes, etc.) because it enables quickly and simply identification of captured anopheles from the DNA contained in their legs. The rest of the mosquito is tested by circumsporozoite protein antigen ELISA and blood meal ELISA. The data obtained are used to determine distribution, cycles, trophic preferences and comparative vectorial capacities of Anopheles gambiae, Anopheles arabiensis and Anopheles melas. -Identification in a mosquito blood meal of the individual bitten: we propose to evaluate factors (weight, age, sex, location of bedroom, etc.) which could explain why individuals are more, or less, bitten by a Plasmodium vector. Genetic typing is used on inhabitants'leukocytes DNA and on the leukocyte DNA extracted from the blood meal of resting anopheles. Through the high degree of polymorphism of three (AAAG)n microsatellites markers, we hope, using PCR, to attribute each blood meal to one individual. Statistical analysis will be used to identify attractivity factors and to determine more precisely the inoculation rates for each group rather than the classical rate calculated with male adults volunteers.

Twenty species of Anopheles are presently known from Senegal. An. gambiae, An. arabiensis, An. pharoensis, An. rufipes and An. ziemanni have an extensive distribution. Probably because of climatic change, An. funestus is no more found in some areas and An. paludis tend to disappear. An. melas is located in coastal areas. The other species, namely An. coustani, An. brohieri, An. brunnipes, An. domicola, An. flavicosta, An. freetownensis, An. hancocki, An. maculipalpis, An. nili, An. pretoriensis, An. squamosus and An. wellcomei, are mainly found in southern Senegal. Only An. gambiae, An. arabiensis and An. funestus are of epidemiological significance as vectors of malaria and bancroftian filariasis. Twelve arboviruses have been isolated from eight Anopheles species. Each Anopheles species is illustrated and an identification key is given.

A survey has been carried out during eight months in the hospital of Bobo-Dioulasso (Burkina Faso) in view to observe the infection of the placentas by Plasmodium and their risk factors. One thousand forty pregnant women were included. Infection rates were 6.5% during the low transmission season and 24.5% during the high transmission season. P. falciparum was present in all the infections. Thirty eight per cent of the women under 18 years old and 26% of the primigravidae were infected. The mean difference in birth weights between the newborn babies of primigravidae with infected or non-infected placentas was 275 g. The regular visits to antenatal clinics, a high level of education and sufficient income were linked with a low level of placental infection. The use of bed nets was linked with a low level of placental infections even after suppression of the other socio-economic data. These observations are discussed in view of an action against malaria in endemic areas.

During November and December 1992, the authors surveyed the malaria morbidity and the drugs efficiency in an urban area of Dakar region. Malaria represented 13.68% of the fevers in Dakar district and 12.4% in Pikine. The mean parasitic density was evaluated at 5000/mm3. Good efficiency of antimalarial drugs were noted but chloroquine-resistance is present with 24% of prevalence.

The malarial infectivity of an African village population was tested by selecting a demographically representative sample of individuals for study, regardless of parasitemia or gametocytemia. The infectivity of this population people to laboratory-bred mosquitoes was investigated using membrane feeding techniques. Tests on 322 subjects (greater than four years of age) indicated that approximately 48.4% were capable of infecting mosquitoes. There were similar proportions of infectious individuals among gametocyte carriers (52.5%) and nongametocyte carriers (46.6%). All age groups appeared to contribute equally to this infective reservoir. Most of the infections resulted in low oocyst loads (1.8 oocysts) on the midgut of the positive mosquitoes and only a few mosquitoes per batch were infected (11.5%). A previous entomologic survey estimated 90 infected bites/person/year and a low parity index in Anopheles gambiae (< 60%) as well as in An. funestus (< 40%), the two main malaria vectors in this region. This low parity index could indicate a low life expectancy for infected mosquitoes and could therefore explain an inoculation rare lower than expected considering the high degree of infectivity of the human population studied.

The gene frequencies of haemoglobin A (HbA), HbS and HbC were studied in Burkina-Faso (BF) and in a neighbouring region of Niger, Ayorou. The frequency of HbS was higher in the Sahel region, (northern part of BF and Ayorou) than in the Savanna region. The reverse was true for the HbC gene. The major findings of this study are: (a) confirmation of a peak of HbC gene frequency in the central region of BF (Mossi plateau); (b) a possible negative correlation between the frequencies of HbS and HbC-Cavalli-Sforza and Bodmer have observed that this correlation is at a significantly different level from that expected because of the allelic relationship between HbS and HbC; (c) comparison with the data collected by Livingstone shows a modification in the fitness of the different genotypes in the last thirty years: AS individuals have a lower and AA and SS a higher fitness. Our data favour a partial selection relaxation in this region.

A retrospective study of 1,003 children permits us to identify precisely rates and causes of mortality in children under 5 years old in a rural holoendemic malaria area of the People's Republic of the Congo. The mortality rates are distinctly lower than those generally observed in tropical Africa. Infant mortality (0-1 year) was found to be 71%, mortality 13%, early neonatal mortality (0-7 days) 41% and mortality between 1-5 years, 49%. Main causes of mortality are those related to pregnancy (prematurity, obstetrical pathology) and with infectious diseases particularly measles. On the other hand, no death seems to be attributable directly to malaria in this study.

Cryptococcus neoformans, mainly widespread in Occidental Europe, America and South East of Asia, was rarely described in African pathology. In Central Africa, it was isolated several times in Zaïre. All reported cases concern young people and present no associated pathology. A first fatal case, looking like an acute meningitis, was described in 1959, in a 18 years old Congolese girl. This second case discovered in Brazzaville in a 37 years old young man was a fatal encephalitis after a primary pneumopathy.

Freeze-dried and freshly cultured isolates of Plasmodium falciparum from different countries have been examined for variation in six enzymes, GPI, PGD, LDH, GDH, ADA and PEPE, using starch gel electrophoresis. Variant forms of each enzyme occur. For most enzymes, similar forms occur in isolates from different parts of the world, although differences in frequencies of each form are found. LDH-2 has been found only in African isolates. PEPE variants vary markedly in their frequency between East and West Africa, although further work is needed to confirm this finding.

An epidemic of yellow fever raged during the last three months of 1983 in South East of Upper Volta. It spread on about ten thousand square kilometers, in a bushy savanna area, affecting only populations living in contact with forest galleries, belonging especially to the peul ethnical group. The transmission of the virus was effected by sylvatic vectors, essentially Aedes furcifer. Serological tests showed that about 50 % of the population living in contact with forest galleries was affected, that is to say 15.000 to 17.500 people. The average death rate on the whole area was 4 % (800 to 1.700 deaths); the lethality rate was estimated between 6 and 10 % of affected people. On the whole, 54 strains of yellow fever virus were isolated from human blood samples, and 26 strains from batches of mosquitoes. We called this epidemic "intermediate sylvatic epidemic". The epidemic quickly decreased in the sylvatic area, owing to climatic conditions. A mass campaign of vaccinations prevented it from spreading to near urban centres. On this particular case, the thermostability on field of the vaccine 17D provided by the Institute Pasteur of Dakar was proved to be effective.

From 1978 to 1979, 5 surveys, among schoolchildren, were carried out during the rainy season in the neighbourhood of Brazzaville (R.P. Congo): 3 in PK 45 village (northern part of the capital), 2 in Djoumouna village (southern part), and 1 in "Talangai" (a suburb of the capital). 868 exams (plasmodic and splenic index fitted with hemoglobin composition [Hb AA or Hb AS]) were done. It appeared that 19,6% of schoolchildren examined were heterozygous sicklers (AS). This percentage confirmed the previous results from other authors in different countries of Central Africa. On the other hand, in spite of an intense transmission, both plasmodic and splenic index were, on the average, relatively low (24,5 and 24,8% respectively). Plasmodium falciparum was largely predominant (95,3% of infections) but P. ovale and P. malariae were also found (1,9% for each species). From our study no obvious "protecting effect" can be attributed to sickle cell trait because plasmodic index of children AA and AS were similar (23,8 and 27,6% respectively). A slight decrease of splenic index was noticed in AS in regard to AA (19,4 and 26.1% respectively). It is difficult to consider this no significative regression as a definitive proof of a premunition stronger in AS than in AA. Effectively some splenic infarctus are well known to be a regular physiopathological process occurring in homozygous SS but often in heterozygous AS too. In such highly endemic and stable malaria area the problem of a suitable antimalaria strategy remains to be solved.

During a study on malaria in pregnant women in Burkina Faso, the authors gave a particular attention to the hemoglobin of a mother and her new-born child (blood of cord) and they noticed an hemoglobin migrating before the HbA which was identified by isoelectric focusing (IEF). The child is a composite HbK/HbC heterozygote. A survey was carried out to check the transmission of such a K Woolwich hemoglobin within the family of the mother. Out of 40 people, 17 got HbKw. A noticeable anemia was found in HbKw/HbC heterozygote. The authors tried to identified a possible thalassemia. There was little probability for an association of a minor alpha-thalassemia in the absence of Bart's hemoglobin in the blood of the cord (IEF test) and there was no associated beta-thalassemia.