Village Of Dielmo Research Articles

The Application of Culturomics to Explore African Skin Microbiota.

Over the past 12 years, culturomics, a high-throughput culture method, has been developed, considerably widening the repertoire of known cultured bacteria. An exhaustive database, including a list of microbes isolated by culture from human skin, was recently established by performing a review of the literature. The aim of the present study was to use the culturomics approach to explore the African skin microbiota. Skin swabs from the palms of human hands were collected between January and December 2016 from healthy subjects from the villages of Dielmo and Ndiop in rural Senegal. Three culture media were selected for the isolation of bacteria in aerobic conditions. Bacterial colonies were subjected to matrix-assisted laser desorption ionization-time of flight mass spectroscopy and the 16 S rRNA gene was sequenced for unidentified colonies. A total of 176 bacterial species were isolated. This increased the repertoire of bacterial species on the skin by 14.0%, by adding 71 bacteria, including seven new species. The culturomics approach characterizing microbial diversity has significantly changed our view of the skin microbiota, raising many important questions about the host-microorganism relationship and its relevance to skin diseases. In particular, the difference between the palm microbiota of these African populations (composed mainly of the genera Staphylococcus, Arthrobacter, Bacillus, and Microbacterium) and that of Western populations, whose main genera are Staphylococcus, Propionibacterium, Micrococcus, Corynebacterium, Enhydrobacter, and Streptococcus. This study demonstrates the need to continue to explore the skin microbiome using the culturomics approach.

Persistent carriage of subpatent Plasmodium falciparum parasites associated with clinical malaria in a low transmission area in Senegal

ObjectivesIn low malaria transmission areas, the elimination of the disease has been hampered partly by the existence of a reservoir of subpatent Plasmodium falciparum infections within communities. This reservoir, often undetected, serves as a source of parasites and contributes to ongoing transmission and clinical malaria cases. MethodsThis study, spanning a period of 9 years from June 2014 to December 2022, examined individual variations and long-term subpatent P. falciparum carriage in two matched cohorts of 44 individuals each living in Dielmo village in the Sudanian area of Senegal. Biannual blood samples, collected in June/July and December of each year, underwent P. falciparum diagnosis by quantitative polymerase chain reaction. QGIS and R analytical tools were used to map infections, assess the occurrence and clustering of subpatent and clinical P. falciparum infections, and determine the risk of infection in the vicinity of asymptomatic P. falciparum carriers. ResultsThe point frequency and long-term P. falciparum carriage were significantly higher among the quantitative polymerase chain reaction (qPCR) positive cohort compared to the negative cohort across the 16 cross-sectional surveys analyzed in this study (19.76% vs 10.99%, P-value <0.001). Asymptomatic carriage events in qPCR-positive group were 18.86 ± 1.72% and significantly greater (P-value = 0.001) than in the qPCR-negative group (11.32 ± 1.32%). The relative risk of P. falciparum infection or clinical malaria calculated with a 95% confidence interval significantly increased in the vicinity of infected individuals and was 1.44 (P-value = 0.53) and 2.64 (P-value = 0.04) when at least one individual in the direct (household) or indirect (block of households) vicinity is infected, respectively. The risk increased to 3.64 (P-value <0.001) if at least 1/5 of individuals in the indirect vicinity were P. falciparum-infected. ConclusionsThe study provides a detailed qualitative and quantitative analysis of the asymptomatic P. falciparum reservoir and its temporal and spatial dynamics within two subgroups of P. falciparum-infected and non-infected individuals in Dielmo village. It identified high-risk populations known as “hotpops” and hotspots that could be targeted by innovative interventions to accelerate the elimination of malaria in Dielmo village.

One hundred malaria attacks since birth. A longitudinal study of African children and young adults exposed to high malaria transmission

Despite significant progress in malaria control over the past twenty years, malaria remains a leading cause of child morbidity and mortality in Tropical Africa. As most patients do not consult any health facility much uncertainty persists about the true burden of the disease and the range of individual differences in susceptibility to malaria. Over a 25-years period, from 1990 to 2015, the inhabitants of Dielmo village, Senegal, an area of intense malaria transmission, have been monitored daily for their presence in the village and the occurrence of diseases. In case of fever thick blood films were systematically examined through microscopy for malaria parasites and patients received prompt diagnosis and treatment. We analysed data collected in 111 children and young adults monitored for at least 10 years (mean 17.3 years, maximum 25 years) enrolled either at birth (95 persons) or during the two first years of life. A total of 11,599 episodes of fever were documented, including 5268 malaria attacks. The maximum number of malaria attacks in a single person was 112. Three other persons suffered one hundred or more malaria attacks during follow-up. The minimum number of malaria attacks in a single person was 11. The mean numbers of malaria attacks in children reaching their 4th, 7th, and 10th birthdays were 23.0, 37.7, and 43.6 attacks since birth, respectively. Sixteen children (14.4%) suffered ten or more malaria attacks each year at ages 1-3 years, and six children (5.4%) each year at age 4-6 years. Long-term close monitoring shows that in highly endemic areas the malaria burden is higher than expected. Susceptibility to the disease may vary up to 10-fold, and for most children childhood is an endless history of malaria fever episodes. No other parasitic, bacterial or viral infection in human populations has such an impact on health. The Pasteur Institutes of Dakar and Paris, the Institut de Recherche pour le Développement, and the French Ministry of Cooperation provided funding.

Rebound of multiple infections and prevalence of anti-malarial resistance associated markers following malaria upsurges in Dielmo village, Senegal, West Africa

BackgroundThanks to the scale up of malaria control interventions, the malaria burden in Senegal has decreased substantially to the point that the National Malaria Control Programme plans to achieve malaria elimination by 2030. To guide such efforts, measuring and monitoring parasite population evolution and anti-malarial drugs resistance is extremely important. Information on the prevalence of parasite mutations related to drug resistance can provide a first signal of emergence, introduction and selection that can help with refining drug interventions. The aim of this study was to analyse the prevalence of anti-malarial drug resistance-associated markers before and after the implementation of artemisinin-based combination therapy (ACT) from 2005 to 2014 in Dielmo, a model site for malaria intervention studies in Senegal.MethodsSamples from both malaria patients and Plasmodium falciparum asymptomatic carriers were analysed with high resolution melting (HRM) technique to genotype P. falciparum chloroquine resistance transporter (Pfcrt) gene haplotypes and multidrug-resistant protein 1 (Pfmdr1) gene at codons N86 and Y184.ResultsAmong the 539 samples analysed, 474, 486, and 511 were successfully genotyped for Pfmdr1 N86, Y184, and Pfcrt, respectively. The prevalence of drug resistance markers was high, particularly during the malaria upsurges. Following the scale-up in bed net distribution, only the mutant (86F-like) variant of Pfmdr1 86 was present while during the malaria upsurges the predominance of two types 86Y-86N (43%) and 86F-like (56%) were observed. Most infections (87%) carried the wild type Y-allele at Pfmdr1 184 during the period of nets scale-up while during the malaria upsurges only 16% of infections had wild type and 79% of infections had mixed (mutant/wild) type. The frequency of the mixed genotypes SVMNT-like_CVMNK and SVMNT-like_CVIET within Pfcrt gene was particularly low during bednet scale up. Their frequency increased significantly (P < 0.001) during the malaria upsurges.ConclusionThis data demonstrated the effect of multiple interventions on the dynamics of drug resistance-associated mutations in the main malaria parasite P. falciparum in an endemic village in Senegal. Monitoring drug resistance markers should be conducted periodically to detect threats of emergence or resurgence that could compromise the efficacy of anti-malarial drugs.

Fine-scale Spatiotemporal Mapping of Asymptomatic and Clinical Plasmodium falciparum Infections: Epidemiological Evidence for Targeted Malaria Elimination Interventions.

A detailed understanding of the contribution of the asymptomatic Plasmodium reservoir to the occurrence of clinical malaria at individual and community levels is needed to guide effective elimination interventions. This study investigated the relationship between asymptomatic Plasmodium falciparum carriage and subsequent clinical malaria episodes in the Dielmo and Ndiop villages in Senegal. The study used a total of 2792 venous and capillary blood samples obtained from asymptomatic individuals and clinical malaria datasets collected from 2013 to 2016. Mapping, spatial clustering of infections, and risk analysis were performed using georeferenced households. High incidences of clinical malaria episodes were observed to occur predominantly in households of asymptomatic P falciparum carriers. A statistically significant association was found between asymptomatic carriage in a household and subsequent episode of clinical malaria occurring in that household for each individual year (P values were 0.0017, 6 × 10-5, 0.005, and 0.008 for the years 2013, 2014, 2015, and 2016 respectively) and the combined years (P = 8.5 × 10-8), which was not found at the individual level. In both villages, no significant patterns of spatial clustering of P falciparum clinical cases were found, but there was a higher risk of clinical episodes <25 m from asymptomatic individuals in Ndiop attributable to clustering within households. The findings provide strong epidemiological evidence linking the asymptomatic P falciparum reservoir to clinical malaria episodes at household scale in Dielmo and Ndiop villagers. This argues for a likely success of a mass testing and treatment intervention to move towards the elimination of malaria in the villages of Dielmo and Ndiop.

Cross-Reactivity of SARS-CoV-2 Laboratory Diagnostics to Endemic Diseases in Africa: A Diagnostic Accuracy Study

Background: Serology is a great tool to assess the level of immunity against SARS-CoV-2 in settings with limited access to molecular diagnostics. However, African populations displays a particular immunological profile with massive circulation of infectious agents from different aetiologies that can affect assays performance.Methods: We evaluated the OMEGA Diagnostics COVID-19 ELISA-IgG and the ID Screen® SARS-CoV-2-N IgG Indirect in Senegal using a panel of 636 blood samples covering several African-endemic diseases and healthy donors to determine test sensitivity and specificity. The sensitivity panel of sera includes 461 serum samples collected from 91 patients hospitalized for COVID-19 disease. COVID-19 cases were confirmed by qRT-PCR and samples were collected on an interval of three days until viral clearance. In addition, 272 sera obtained from COVID-19 negative individuals were selected from a well-documented biobank of sera collected before the COVID-19 outbreak.Finding: High-cross reactivity have been found in individuals with a history of exposure to Chikungunya, HIV, malaria (Plasmodium falciparum), rheumatoid factor as well as healthy donors with respective specificities of 55%, 41.8%, 70%, 70% and 75%. ELISA experiments with commercial assays targeting either SARS-CoV-2 Nucleocapsid protein and Spike 2 protein or nucleocapsid protein only suggest that cross-reactivity might be directed against Spike 2 protein and not Nucleocapsid protein. Further samples characterisation reveals that anti-malaria IgG is the leading cause of such poor specificities, but exposure to other diseases contributed as well.Interpretation: We anticipate that COVID-19 seroprevalence can be biased if assays are not contextualized. Since malaria is endemic in African settings, we propose that a particular attention must be given in serological surveillance of COVID-19 or anti-SARS-CoV-2 antibodies quantification as vaccines are being rolled out.FundingUK Foreign, Commonwealth and Development Office/Wellcome Trust Joint Initiative for Research in Epidemic Preparedness and Response (JIREP grant number 220764/Z/20/Z).Funding Information: UK Foreign, Commonwealth and Development Office/Wellcome Trust Joint Initiative for Research in Epidemic Preparedness and Response (JIREP grant number 220764/Z/20/Z).Declaration of Interests: JRAF was an employee of Mologic Ltd, which was the development partner of one of the ELISAs adopted in this study. The remaining authors declare no competing interest.Ethics Approval Statement: Pre-COVID-19 samples for malaria (PCR), dengue, yellow fever, Zika, Chikungunya, Influenza A/B, HIV, rheumatoid arthritis and samples tested negative for the same diseases and also Crimean Congo haemorrhagic fever, West Nile fever encephalitis and Rift Valley fever were part of national public health surveillance program of the Senegalese Ministry of Social Action and Health performed in collaboration with Institut Pasteur de Dakar. Therefore, consultation with ethics committee was not required. Pre-COVID-19 samples for malaria endemic areas were from a longitudinal cohort survey performed in Dielmo village and approved by the Senegalese National Ethics Committee for Research in Health (reference number 00000007/MSAS/CNERS/Sec 26 January 2021). Samples from COVID-19 RT-PCR positive patients were obtained from a multicentre cohort survey approved by the Senegalese National Ethics Committee for Research in Health (reference number 00000068/MSAS/CNERS/Sec, 10 April 2020).

The impact of daily soap use in rural areas of Senegal on respiratory infectious diseases, fevers and skin microbiota

ObjectivesChildren aged <5 years are the group most affected by infectious diseases, more specifically in underdeveloped countries. A study was performed to assess the effects of daily soap use on the incidence of diarrhoea, fever, respiratory infection, and the prevalence of pathogenic bacteria on the skin. MethodsSoap was distributed to the population of the village of Ndiop (test) for use in their daily hygiene but not to the population of the village of Dielmo (control). Fieldworkers daily recorded the clinical events in the two villages and encouraged the use of soap in Ndiop. ResultsA total of 638 people participated in the study. The incidence rates of cough, runny nose and fever significantly decreased in 2016 compared with 2015, unlike that of diarrhoea. In 2016, significant reductions in the incidence rates of cough, runny nose and fever were observed in children aged <15 years in Ndiop. The prevalence of Streptococcus pneumoniae, Staphylococcus aureus and Streptococcus pyogenes in the palms of the hands significantly dropped in Ndiop. ConclusionUsing soap reduces the incidence of respiratory infections, fevers and the prevalence of pathogenic bacteria on the skin. However, for diarrhoea, additional strategies are needed to improve outcomes.

Study of Bacteriological and Parasitological Management Quality of Water in the Villages of Senegal

A survey on the parasitological and bacteriological management was conducted in two villages in southeastern Senegal. The objective of this study was to evaluate the water management modalities in the two villages with ethnic differences and possibly detect factors that could cause waterborne diseases. A questionnaire on water management methods was submitted to one hundred one women drawn in the villages of Dielmo and Ndiop. Samples of well water and water stored in canaries were subjected to both parasitological and bacteriological analysis. In both villages, there was a difference in water management with respect to household configuration (monogamous, polygamous). The containers used for the drawing and storage of water were the same in their two villages. No helminth eggs were found in the drinking water, but bacterial germs, especially faecal coliforms (an indicator of human contamination) and Enterocoques (an indicator of animal contamination). These waters do not comply with WHO standards. So it is important to disinfect them before consumption although no cases of gastroenteritis have been reported.

Retraction: How the Malaria Vector Anopheles gambiae Adapts to the Use of Insecticide-Treated Nets by African Populations.

Background Insecticide treated bed nets have been recommended and proven efficient as a measure to protect African populations from malaria mosquito vector Anopheles spp. This study evaluates the consequences of bed nets use on vectors resistance to insecticides, their feeding behavior and malaria transmission in Dielmo village, Senegal, were LLINs were offered to all villagers in July 2008. Methods Adult mosquitoes were collected monthly from January 2006 to December 2011 by human landing catches (HLC) and by pyrethroid spray catches (PCS). A randomly selected sub-sample of 15–20% of An. gambiae s.l. collected each month was used to investigate the molecular forms of the An. gambiae complex, kdr mutations, and Plasmodium falciparum circumsporozoite (CSP) rate. Malaria prevalence and gametocytaemia in Dielmo villagers were measured quarterly. Results Insecticide susceptible mosquitoes (wild kdr genotype) presented a reduced lifespan after LLINs implementation but they rapidly adapted their feeding behavior, becoming more exophageous and zoophilic, and biting earlier during the night. In the meantime, insecticide-resistant specimens (kdr L1014F genotype) increased in frequency in the population, with an unchanged lifespan and feeding behaviour. P. falciparum prevalence and gametocyte rate in villagers decreased dramatically after LLINs deployment. Malaria infection rate tended to zero in susceptible mosquitoes whereas the infection rate increased markedly in the kdr homozygote mosquitoes. Conclusion Dramatic changes in vector populations and their behavior occurred after the deployment of LLINs due to the extraordinary adaptative skills of An. gambiae s. l. mosquitoes. However, despite the increasing proportion of insecticide resistant mosquitoes and their almost exclusive responsibility in malaria transmission, the P. falciparum gametocyte reservoir continued to decrease three years after the deployment of LLINs.

Acute Febrile Illness and Influenza Disease Burden in a Rural Cohort Dedicated to Malaria in Senegal, 2012-2013.

BackgroundAfrican populations are considered to be particularly vulnerable to fever illnesses, including malaria, and acute respiratory disease, owing to limited resources and overcrowding. However, the overall burden of influenza in this context is poorly defined and incidence data for African countries are scarce. We therefore studied the fever syndrome incidence and more specifically influenza incidence in a cohort of inhabitants of Dielmo and Ndiop in Sokone district, Senegal.MethodsDaily febrile-illness data were prospectively obtained from January 2012 to December 2013 from the cohort of the villages of Dielmo and Ndiop, initially dedicated to the study of malaria. Nasopharyngeal swabs were collected from, and malaria diagnosis tests (thick blood smears) carried out on, every febrile individual during clinical visits; reverse transcriptase-polymerase chain reaction was used to identify influenza viruses in the samples. Binomial negative regression analysis was used to study the relationship between the monthly incidence rate and various covariates.ResultsIn Dielmo and Ndiop, the incidence of malaria has decreased, but fever syndromes remain frequent. Among the 1036 inhabitants included in the cohort, a total of 1,129 episodes of fever were reported. Influenza was present all year round with peaks in October-December 2012 and August 2013. The fever, ILI and influenza incidence density rates differed significantly between age groups. At both sites, the adjusted incidence relative risks for fever syndromes and ILI were significantly higher in the [6–24 months) than other age groups: 7.3 (95%CI: [5.7–9.3]) and 16.1 (95%CI: [11.1–23.3]) respectively. The adjusted incidence relative risk for influenza was significantly higher for the [0–6 months) than other age groups: 9.9 (95%CI: [2.9–33.6]). At both sites, incidence density rates were lowest among adults > = 50 years.ConclusionsIn this rural setting in Senegal, influenza was most frequent among the youngest children. Preventive strategies targeting this population should be implemented.

The value of local malaria strains for serological studies: local strains versus Palo Alto reference strain.

BackgroundThe standardization of the type of crude Plasmodium falciparum extracts for assays to evaluate the overall anti-blood-stage immune response in humans may be beneficial to malaria pre-elimination programmes. However, there is no consensus on which strain is appropriate for routine analyses. This study aimed to compare the responses of malaria IgG antibodies in serum collections from Dielmo and Ndiop to crude extracts of merozoites and schizonts of local and reference strains of P. falciparum.MethodsMalaria antibodies were evaluated using serological tests for exposure to three local strains (0703, F15 and F16) and the P. falciparum reference Palo Alto strain (PA). A total of 218 sera collected in 2000 from inhabitants of the villages of Dielmo and Ndiop were used: 142 from Dielmo and 76 from Ndiop. The biological collection was used to evaluate by ELISA the prevalence of IgG antibodies against crude merozoite and schizont extracts. The genetics of the local and reference strains were compared.ResultsThere was genetic divergence between strains 0703, F15, F16 and PA. IgG responses against local and reference strains correlated well (0.6 to 0.8; p < 0.01). Ig G responses were highest to schizont and merozoite extracts from the field strain of P. falciparum 0703 adapted to in vitro culture. Extracts of P. falciparum strain 0703 isolated from a subject in Dielmo was the most widely recognized [91.3 % (199/218) and 81.2 % (177/218) for schizonts and merozoites, respectively], although the responses were high for merozoites from PA [85.3 % (186/218)] the reference strain, and the two strains isolated from subjects living in Dakar: F15 [90.4 % (197/218)] and F16 [72.5 % (158/218)].ConclusionsFor serological studies, the local strain provided the most complete picture of exposure to transmission and malaria prevalence in the local context. However, for the standardization of this method by different laboratories, the reference strain appeared to perform sufficiently well to be used for the evaluation of malaria control programmes.

The implication of long-lasting insecticide-treated net use in the resurgence of malaria morbidity in a Senegal malaria endemic village in 2010-2011.

BackgroundAlthough the burden of malaria has significantly declined in recent years in sub-Saharan Africa through the widespread use of long-lasting insecticide treated bed-nets (LLINs) and artemisinin-based combination therapy, resurgence of malaria is observed in some settings after several years of LLINs use. This study aimed to assess if LLINs use remains protective against malaria during a period of resurgence of malaria morbidity in Dielmo, a rural village of Senegal.MethodsIn July 2008, LLINs were offered to all villagers and lately in July 2011, LLINs were renewed.A longitudinal study was conducted between July, 2010 and December, 2011 among inhabitants of the village of Dielmo to identify all episodes of fever. Thick smears stained with Giemsa were done for every febrile villager and malaria attacks were treated with combination of Artesunate plus Amodiaquine. Cross-sectional surveys were also conducted at the end of the rainy season (October 2010 and November 2011) to assess asymptomatic carriage. A survey on LLINs use was done every quarter of the year. A random-effect logistic regression was used to assess the effect of LLINs use on the risk of having a malaria attack after adjusting for the main risk factors.ResultsThe study population included 449 individuals corresponding to a total of 2140 observations. One hundred and fifteen (115) clinical malaria attacks attributed to P. falciparum (cases) have been recorded over the study period. Most of the malaria cases occurred in October-December 2010 (49/115 i.e. 43%) and among adults aged 15 years and over (50/115, i.e. 43%). During the study period, the use of LLINs was 61% among non-malaria cases and only 42% among malaria clinical cases but differenced according to age group.After adjusting on gender, age, rainfall and LLINs replacement, we found that LLINs use (AOR [95%CI] = 0.40 [0.25; 0.62], p < 0.001) remained a protective factor against malaria attacks during the study period.ConclusionLLINs use remains effective to reduce malaria burden. These results highlight the need to pursue LLINs implementation in the current context of malaria elimination and to provide positive incentives to increase its use in the population.

No difference in the incidence of malaria in human-landing mosquito catch collectors and non-collectors in a Senegalese village with endemic malaria.

BackgroundThe human landing catches is the gold standard method used to study the vectors of malaria and to estimate their aggressiveness. However, this method has raised safety concerns due to a possible increased risk of malaria or other mosquito-borne diseases among the mosquito collectors. The aim of this study was to evaluate the incidence of malaria attacks among mosquito collectors and to compare these results with those of non-collectors in a Senegalese village.MethodsFrom July 1990 to December 2011, a longitudinal malaria study involving mosquito collectors and non-collectors was performed in Dielmo village, Senegal. During the study period, 4 drugs were successively used to treat clinical malaria, and long-lasting insecticide-treated nets were offered to all villagers in July 2008. No malaria chemoprophylaxis was given to mosquito collectors. Incidence of uncomplicated clinical malaria and asymptomatic malaria infection were analyzed among these two groups while controlling for confounding factors associated with malaria risk in random effects negative binomial and logistic regression models, respectively.ResultsA total of 3,812 person-trimester observations of 199 adults at least 15 years of age were analyzed. Clinical malaria attacks accounted for 6.3% both in collectors and non-collectors, and asymptomatic malaria infections accounted for 21% and 20% in collectors and non-collectors, respectively. A non-significant lower risk of malaria was observed in the collector group in comparison with the non-collector group after adjusting for other risk factors of malaria and endemicity level (Clinical malaria: adjusted incidence rate ratio = 0.89; 95% confidence interval = 0.65-1.22; p= 0.47).ConclusionBeing a mosquito collector in Dielmo was not significantly associated with an increased risk of malaria both under holoendemic, mesoendemic and hypoendemic conditions of malaria epidemiology. This result supports the view that HLC, the most accurate method for evaluating malaria transmission, may be used without health concerns in Dielmo.

How the malaria vector Anopheles gambiae adapts to the use of insecticide-treated nets by African populations.

BackgroundInsecticide treated bed nets have been recommended and proven efficient as a measure to protect African populations from malaria mosquito vector Anopheles spp. This study evaluates the consequences of bed nets use on vectors resistance to insecticides, their feeding behavior and malaria transmission in Dielmo village, Senegal, were LLINs were offered to all villagers in July 2008.MethodsAdult mosquitoes were collected monthly from January 2006 to December 2011 by human landing catches (HLC) and by pyrethroid spray catches (PCS). A randomly selected sub-sample of 15–20% of An. gambiae s.l. collected each month was used to investigate the molecular forms of the An. gambiae complex, kdr mutations, and Plasmodium falciparum circumsporozoite (CSP) rate. Malaria prevalence and gametocytaemia in Dielmo villagers were measured quarterly.ResultsInsecticide susceptible mosquitoes (wild kdr genotype) presented a reduced lifespan after LLINs implementation but they rapidly adapted their feeding behavior, becoming more exophageous and zoophilic, and biting earlier during the night. In the meantime, insecticide-resistant specimens (kdr L1014F genotype) increased in frequency in the population, with an unchanged lifespan and feeding behaviour. P. falciparum prevalence and gametocyte rate in villagers decreased dramatically after LLINs deployment. Malaria infection rate tended to zero in susceptible mosquitoes whereas the infection rate increased markedly in the kdr homozygote mosquitoes.ConclusionDramatic changes in vector populations and their behavior occurred after the deployment of LLINs due to the extraordinary adaptative skills of An. gambiae s. l. mosquitoes. However, despite the increasing proportion of insecticide resistant mosquitoes and their almost exclusive responsibility in malaria transmission, the P. falciparum gametocyte reservoir continued to decrease three years after the deployment of LLINs.

Genetic polymorphism at an odorant receptor gene (Or39) among mosquitoes of the Anopheles gambiae complex in Senegal (West Africa).

BackgroundOlfaction plays a significant role in insect behavior during critical steps of their life-cycle, such as host-seeking during foraging or the search for a mate. Here, we explored genetic polymorphism within and divergence between sibling species of the African malaria mosquito, Anopheles gambiae sensu lato in the gene sequence and encoded peptides of an odorant receptor, Or39. This study included sympatric specimens of An. gambiae sensu stricto, An. coluzzii and An. arabiensis sampled together in the village of Dielmo, Senegal.ResultsA 1,601 bp genomic sequence composed of 6 exons and 5 introns was obtained for Or39 from 6–8 mosquitoes in each of the 3 species. DNA sequence analysis revealed a high level of molecular polymorphism (π = 0.0154; Haplotype diversity = 0.867) and high overall genetic differentiation between taxa (Fst > 0.92, P < 0.01). In total, 50 parsimony informative sites were recorded. Throughout the whole dataset, there were 13 non-synonymous mutations resulting in aminoacid changes in the encoded protein. Each of the 6 different identified peptides was species-specific and none was shared across species. Most aminoacid changes were located on the intracellular domains of the protein. However, intraspecific polymorphisms in An. gambiae and An. arabiensis as well as species-specific mutations also occurred in the first extracellular domain.ConclusionsAlthough obtained from a limited number of specimens, our results point towards genetic differences between cryptic species within the An. gambiae complex in a gene of biological relevance that might be of evolutionary significance when exposed to disruptive selective forces.

Rickettsia felis and related bacteria: An epidemiological enigma

Background: Recent works identified a novel healthcare problem in Africa: acute Rickettsia felis-induced fever that mimics malaria. The epidemiology of these diseases remains to be characterized. Indeed, the natural reservoirs of R. felis are not known. Since the first clinical descriptions of R. felis-associated fever, the cat and dog fleas Ctenocephalides felis and C. canis were implicated as themost probable vectors inmany countries, but not in Senegal. Methods & Materials: Arthropods (ticks, mites, fleas, tsetse flies, biting midges, mosquitoes, bed bugs) and environmental samples (dust and water samples) were collected in the villages of Dielmo and Ndiop where R. felis-associated acute fevers contribute up to 8% of all acute fevers with the incidence of 6.7 per 100 person-years. DNA extracted from all samples was tested by Rickettsia genus-specific qPCR and by two different R. felis-specific qPCR. Positive environmental samples were subjected to PCR with the universal eukaryotic cytochrome oxydase I-based primers followedbycloningof theampliconswithpGemvectors. Isolationwas performed in XTC cell line using the shell-vial technique. Characterization of the isolated strainswas done by sequencing of gltA and 16S rRNA genes. Results: Arthropods (ticks, mites, fleas, tsetse flies, biting midges, mosquitoes, bed bugs) and environmental samples (dust and water samples) were collected in the villages of Dielmo and Ndiop where R. felis-associated acute fevers contribute up to 8% of all acute fevers with the incidence of 6.7 per 100 person-years. DNA extracted from all samples was tested by Rickettsia genusspecific qPCR and by two different R. felis-specific qPCR. Positive environmental samples were subjected to PCR with the universal eukaryotic cytochrome oxydase I (COI)-based primers followed by cloning of the amplicons with pGem vectors. Isolation was performed in XTC cell line using the shell-vial technique. Characterization of the isolated strainswas done by sequencing of gltA and 16S rRNA genes. Conclusion: This is the first evidence of the presence of R. felis in arthropods in the endemic focus or R. felis infection in Senegal. The roles of Siteroptidae mites in the hosting and transmission of the R. felis is to be further investigated

A 20-Year Longitudinal Study of Plasmodium ovale and Plasmodium malariae Prevalence and Morbidity in a West African Population

Background Plasmodium ovale and Plasmodium malariae have long been reported to be widely distributed in tropical Africa and in other major malaria-endemic areas of the world. However, little is known about the burden caused by these two malaria species.Methods and FindingsWe did a longitudinal study of the inhabitants of Dielmo village, Senegal, between June, 1990, and December, 2010. We monitored the inhabitants for fever during this period and performed quarterly measurements of parasitemia. We analyzed parasitological and clinical data in a random-effect logistic regression model to investigate the relationship between the level of parasitemia and the risk of fever and to establish diagnostic criteria for P. ovale and P. malariae clinical attacks. The prevalence of P. ovale and P. malariae infections in asymptomatic individuals were high during the first years of the project but decreased after 2004 and almost disappeared in 2010 in relation to changes in malaria control policies. The average incidence densities of P. ovale and P. malariae clinical attacks were 0.053 and 0.093 attacks per person per year in children <15 years and 0.024 and 0.009 attacks per person per year in adults ≥15 years, respectively. These two malaria species represented together 5.9% of the malaria burden.Conclusions P. ovale and P. malariae were a common cause of morbidity in Dielmo villagers until the recent dramatic decrease of malaria that followed the introduction of new malaria control policies. P. ovale and P. malariae may constitute an important cause of morbidity in many areas of tropical Africa.

Pattern of antibodies to the Duffy binding like domain of Plasmodium falciparum antigen Pf332 in Senegalese individuals

Acquisition of antibodies against blood stage antigens is crucial in malaria immunity and the Plasmodium falciparum antigen Pf332, which is present in close association with the infected red blood cell membrane, is one such antigen. In this study, the antibody response to a Duffy binding like fragment of Pf332, termed Pf332-DBL was investigated in sera from naturally exposed individuals living in Dielmo village, Senegal, with regard to immunoglobulin classes (IgG, IgM, IgE) and IgG subclasses (IgG1–4). While the levels of IgM, IgG, IgG1 and IgG2 only displayed a moderate trend to increase with age, Pf332-DBL specific IgG3 levels increased significantly in the older villagers. In multivariate analysis, when controlling for confounding factors, and in a linear model with a Poisson distribution, anti-Pf332-DBL IgG3 as well as the ratio of cytophilic to non cytophilic anti-Pf332-DBL antibodies were found significantly associated with a reduced risk of malaria attack. This association was also present when the IgG3:IgG1 ratio was tested. Finally, two subgroups of villagers with the same mean age, were delineated by IgG3 concentrations either lower or higher than the median value. A total of 45.2% of the individuals with low anti-Pf332-DBL-IgG3 levels but only 21.4% of the villagers in the group with high levels of such antibodies had a clinical malaria attack during a period of 3 years of continuous follow-up after the blood sampling. In conclusion, Pf332-DBL induces naturally the acquisition of antibodies, and Pf332-DBL-specific IgG3 appears to be associated with protection against malaria in this endemic setting.

A New Rickettsia Species Found in Fleas Collected from Human Dwellings and from Domestic Cats and Dogs in Senegal

The insects of the order Siphonaptera, commonly named fleas, are vectors of pathogens around the world. Our previous studies showed that 4.4% of acute febrile diseases in the Sine-Saloum region of Senegal were due to Rickettsia felis. The aim of this study was to explain the high prevalence of R. felis infections in two rural Senegalese populations by an entomological, systematic monitoring protocol. A total of 232 fleas from three species (Ctenocephalides felis, Echidnophaga gallinacea, and Synosternus pallidus) were collected by candle trapping and manually from pets in the villages of Dielmo and Ndiop during the year 2010. The fleas were then tested for the presence of Bartonella and Rickettsia species. No fleas were found to be positive for any Bartonella species or R. felis. Surprisingly, we found that 91.4% of S. pallidus were infected by a new Rickettsia species, which, based on sequence analysis of gltA, ompB, and two fragments of rpoB, was found to be closely related to R. felis. The results from this study did not explain the high incidence of R. felis infections in these Senegalese populations.

Malaria morbidity and pyrethroid resistance after the introduction of insecticide-treated bednets and artemisinin-based combination therapies: a longitudinal study

Substantial reductions in malaria have been reported in several African countries after distribution of insecticide-treated bednets and the use of artemisinin-based combination therapies (ACTs). Our aim was to assess the effect of these policies on malaria morbidity, mosquito populations, and asymptomatic infections in a west African rural population. We did a longitudinal study of inhabitants of Dielmo village, Senegal, between January, 2007, and December, 2010. We monitored the inhabitants for fever during this period and we treated malaria attacks with artesunate plus amodiaquine. In July, 2008, we offered longlasting insecticide (deltamethrin)-treated nets (LLINs) to all villagers. We did monthly night collections of mosquitoes during the whole study period, and we assessed asymptomatic carriage from cross-sectional surveys. Our statistical analyses were by negative binomial regression, logistic regression, and binomial or Fisher exact test. There were 464 clinical malaria attacks attributable to Plasmodium falciparum during 17,858 person-months of follow-up. The incidence density of malaria attacks averaged 5·45 (95% CI 4·90-6·05) per 100 person-months between January, 2007, and July, 2008, before the distribution of LLINs. Incidence density decreased to 0·41 (0·29-0·55) between August, 2008, and August, 2010, but increased back to 4·57 (3·54-5·82) between September and December, 2010--ie, 27-30 months after the distribution of LLINs. The rebound of malaria attacks were highest in adults and children aged 10 years or older: 45 (63%) of 71 malaria attacks recorded in 2010 compared with 126 (33%) of 384 in 2007 and 2008 (p<0·0001). 37% of Anopheles gambiae mosquitoes were resistant to deltamethrin in 2010, and the prevalence of the Leu1014Phe kdr resistance mutation increased from 8% in 2007 to 48% in 2010 (p=0·0009). Increasing pyrethroid resistance of A gambiae and increasing susceptibility of older children and adults, probably due to decreasing immunity, caused the rebound and age shift of malaria morbidity. Strategies to address the problem of insecticide resistance and to mitigate its effects must be urgently defined and implemented. Institut de Recherche pour le Développement and the Pasteur Institute of Dakar.