clove and Malaria--Vivax

Madagascar is considered as a sub-region of the Afrotropical geographical Region in spite of the high endemicity of 95% of the invertebrates. Nevertheless the three malaria vectors An. gambiae s.s., An. arabiensis and An. funestus are quite similar to those of the continental Africa. This support the hypothesis of their recent introduction. Plasmodium falciparum is the dominant parasite but the prevalence of P. vivax is not negligible. It is linked to the Asian component of the human population. P. malariae and P. ovale are of minor importance. The main epidemiological "facies" of Africa are found in Madagascar. The equatorial facies on the East Coast is characterized by a high transmission all year long. In the tropical facies on the West Coast transmission is seasonal (7 months at least). In both areas, malaria is stable and the inhabitants acquire a high immunity before the age of ten; most of the severe cases touch children below 10. The three vectors can be found but An. gambiae s.s. is dominant. In the exophilic southern facies the transmission is seasonal (two to four months). The only vector is An. arabiensis. Malaria is unstable and severe epidemics occur during the years of high rainfall. All age groups are vulnerable because the population is not immune in the Plateaux facies above 1,000 m., malaria is unstable. Severe epidemics occurred in 1987-1988. The vectors are An. Arabiensis and An. funestus. The occurrence of P. falciparum on the Plateaux seems linked to the development of irrigation of rice farming in the XIXth century. Most of the anopheles breeding places on the Plateaux are dependent on rice cultivation. Urban development has brought the inhabitants of the suburbs in close contact with rice fields. Despite the high number of anopheline bites the number of malaria cases remains by far lower than in the neighbouring rural areas. Regional migrations inside the island bring non-immune populations, from the south and the plateaux, in highly malarious areas of the coast, where the migrants are exposed to high risk. In spite of 40 years of uncontrolled use, chloroquine can still cure most, if not all, of malaria cases. Control measures appropriated to the different areas of Madagascar are discussed.

Topics: Agriculture; Animals; Anopheles; Chloroquine; Emigration and Immigration; Humans; Insect Vectors; Madagascar; Malaria; Malaria, Falciparum; Malaria, Vivax; Oryza; Plasmodium malariae; Population Surveillance; Prevalence; Risk Factors; Seasons; Severity of Illness Index; Urbanization; Water

Malaria is still a heavy public health concern in Madagascar. Few studies combining parasitology and entomology have been conducted despite the need for accurate information to design effective vector control measures. In a Malagasy region of moderate to intense transmission of both Plasmodium falciparum and P. vivax, parasitology and entomology have been combined to survey malaria transmission in two nearby villages.. Community-based surveys were conducted in the villages of Ambohitromby and Miarinarivo at three time points (T1, T2 and T3) during a single malaria transmission season. Human malaria prevalence was determined by rapid diagnostic tests (RDTs), microscopy and real-time PCR. Mosquitoes were collected by human landing catches and pyrethrum spray catches and the presence of Plasmodium sporozoites was assessed by TaqMan assay.. Malaria prevalence was not significantly different between villages, with an average of 8.0% by RDT, 4.8% by microscopy and 11.9% by PCR. This was mainly due to P. falciparum and to a lesser extent to P. vivax. However, there was a significantly higher prevalence rate as determined by PCR at T2 ([Formula: see text] = 7.46, P = 0.025). Likewise, mosquitoes were significantly more abundant at T2 ([Formula: see text] = 64.8, P < 0.001), especially in Ambohitromby. At T1 and T3 mosquito abundance was higher in Miarinarivo than in Ambohitromby ([Formula: see text] = 14.92, P < 0.001). Of 1550 Anopheles mosquitoes tested, 28 (1.8%) were found carrying Plasmodium sporozoites. The entomological inoculation rate revealed that Anopheles coustani played a major contribution in malaria transmission in Miarinarivo, being responsible of 61.2 infective bites per human (ib/h) during the whole six months of the survey, whereas, it was An. arabiensis, with 36 ib/h, that played that role in Ambohitromby.. Despite a similar malaria prevalence in two nearby villages, the entomological survey showed a different contribution of An. coustani and An. arabiensis to malaria transmission in each village. Importantly, the suspected secondary malaria vector An. coustani, was found playing the major role in malaria transmission in one village. This highlights the importance of combining parasitology and entomology surveys for better targeting local malaria vectors. Such study should contribute to the malaria pre-elimination goal established under the 2018-2022 National Malaria Strategic Plan.

Topics: Animals; Anopheles; Disease Vectors; Madagascar; Malaria, Falciparum; Malaria, Vivax; Microscopy; Mosquito Vectors; Plasmodium falciparum; Plasmodium vivax; Polymerase Chain Reaction; Staining and Labeling

Community prevalence of infection is a widely used, standardized metric for evaluating malaria endemicity. Conventional methods for measuring prevalence include light microscopy and rapid diagnostic tests (RDTs), but their detection thresholds are inadequate for diagnosing low-density infections. The significance of submicroscopic malaria infections is poorly understood in Madagascar, a country of heterogeneous malaria epidemiology. A cross-sectional community survey in the western foothills of Madagascar during the March 2014 transmission season found malaria infection to be predominantly submicroscopic and asymptomatic. Prevalence of

Topics: Adolescent; Adult; Asymptomatic Diseases; Child; Child, Preschool; Cross-Sectional Studies; Duffy Blood-Group System; Female; Gene Expression; Health Surveys; Humans; Infant; Madagascar; Malaria, Falciparum; Malaria, Vivax; Male; Microscopy; Plasmodium falciparum; Plasmodium vivax; Polymerase Chain Reaction; Prevalence; Receptors, Cell Surface; Risk Factors; Rural Population

The interaction between Plasmodium vivax Duffy binding protein (PvDBP) and Duffy antigen receptor for chemokines (DARC) has been described as critical for the invasion of human reticulocytes, although increasing reports of P. vivax infections in Duffy-negative individuals questions its unique role. To investigate the genetic diversity of the two main protein ligands for reticulocyte invasion, PvDBP and P. vivax Erythrocyte Binding Protein (PvEBP), we analyzed 458 isolates collected in Cambodia and Madagascar from individuals genotyped as Duffy-positive. First, we observed a high proportion of isolates with multiple copies PvEBP from Madagascar (56%) where Duffy negative and positive individuals coexist compared to Cambodia (19%) where Duffy-negative population is virtually absent. Whether the gene amplification observed is responsible for alternate invasion pathways remains to be tested. Second, we found that the PvEBP gene was less diverse than PvDBP gene (12 vs. 33 alleles) but provided evidence for an excess of nonsynonymous mutations with the complete absence of synonymous mutations. This finding reveals that PvEBP is under strong diversifying selection, and confirms the importance of this protein ligand in the invasion process of the human reticulocytes and as a target of acquired immunity. These observations highlight how genomic changes in parasite ligands improve the fitness of P. vivax isolates in the face of immune pressure and receptor polymorphisms.

Topics: Antigens, Protozoan; Cambodia; Cross-Sectional Studies; Genetic Variation; Genotype; Humans; Madagascar; Malaria, Vivax; Plasmodium vivax; Protozoan Proteins; Receptors, Cell Surface

The country received malaria-free certification from WHO in September 2016, becoming only the second country in the WHO South East Asia region to be declared malaria-free. Imported malaria cases continue to be reported, with 278 cases reported between 2013 and 2017. The diagnosis of a severe Plasmodium vivax patient co-infected with HIV and tuberculosis is discussed with an overview of the rapid response mounted by the Anti Malaria Campaign (AMC), Sri Lanka.. It is important to consider comorbid conditions and immunosuppression when a patient with a benign form of malaria presents with severe manifestations. Measures should be strengthened to prevent importation of diseases, such as malaria and AIDS through migrant workers who return from high-risk countries.

Topics: Adult; Case Management; Coinfection; Communicable Diseases, Imported; HIV Infections; Humans; Madagascar; Malaria, Vivax; Male; Sri Lanka; Travel; Tuberculosis

The prevalence and variants of G6PD deficiency in the Plasmodium vivax-endemic zones of Madagascar remain unknown. The admixed African-Austronesian origins of the Malagasy population make it probable that a heterogeneous mix of genetic variants with a spectrum of clinical severity will be circulating. This would have implications for the widespread use of P. vivax radical cure therapy. Two study populations in the P. vivax-endemic western foothills region of Madagascar were selected for G6PD screening. Both the qualitative fluorescent spot test and G6PD genotyping were used to screen all participants.. A total of 365 unrelated male volunteers from the Tsiroanomandidy, Mandoto, and Miandrivazo districts of Madagascar were screened and 12.9% were found to be phenotypically G6PD deficient. Full gene sequencing of 95 samples identified 16 single nucleotide polymorphisms, which were integrated into a genotyping assay. Genotyping (n = 291) found one individual diagnosed with the severe G6PD Mediterranean. Deployment of P. vivax radical cure in Madagascar must be considerate of the risks presented by the observed prevalence of G6PDd prevalence. The potential morbidity associated with cumulative episodes of P. vivax clinical relapses requires a strategy for increasing access to safe radical cure. The observed dominance of African G6PDd haplotypes is surprising given the known mixed African-Austronesian origins of the Malagasy population; more widespread surveying of G6PDd epidemiology across the island would be required to characterize the distribution of G6PD haplotypes across Madagascar.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Diagnostic Tests, Routine; Endemic Diseases; Genotype; Genotyping Techniques; Glucosephosphate Dehydrogenase Deficiency; Humans; Infant; Infant, Newborn; Madagascar; Malaria, Vivax; Male; Mass Screening; Middle Aged; Prevalence; Sequence Analysis, DNA; Young Adult

Deforestation and land-use change have the potential to alter human exposure to malaria. A large percentage of Madagascar's original forest cover has been lost to slash-and-burn agriculture, and malaria is one of the top causes of mortality on the island. In this study, the influence of land-use on the distribution of Plasmodium vectors and concomitant Plasmodium infection in humans and mosquito vectors was examined in the southeastern rainforests of Madagascar.. From June to August 2013, health assessments were conducted on individuals living in sixty randomly selected households in six villages bordering Ranomafana National Park. Humans were screened for malaria using species-specific rapid diagnostic tests (RDTs), and surveyed about insecticide-treated bed net (ITN) usage. Concurrently, mosquitoes were captured in villages and associated forest and agricultural sites. All captured female Anopheline mosquitoes were screened for Plasmodium spp. using a circumsporozoite enzyme-linked immunosorbent assay (csELISA).. Anopheles spp. dominated the mosquito communities of agricultural and village land-use sites, accounting for 41.4 and 31.4 % of mosquitoes captured respectively, whereas Anopheles spp. accounted for only 1.6 % of mosquitoes captured from forest sites. Interestingly, most Anopheles spp. (67.7 %) were captured in agricultural sites in close proximity to animal pens, and 90.8 % of Anopheles mosquitoes captured in agricultural sites were known vectors of malaria. Three Anopheline mosquitoes (0.7 %) were positive for malaria (Plasmodium vivax-210) and all positive mosquitoes were collected from agricultural or village land-use sites. Ten humans (3.7 %) tested were positive for P. falciparum, and 23.3 % of those surveyed reported never sleeping under ITNs.. This study presents the first report of malaria surveillance in humans and the environment in southeastern Madagascar. These findings suggest that even during the winter, malaria species are present in both humans and mosquitoes; with P. falciparum found in humans, and evidence of P. vivax-210 in mosquito vectors. The presence of P. vivax in resident vectors, but not humans may relate to the high incidence of humans lacking the Duffy protein. The majority of mosquito vectors were found in agricultural land-use sites, in particular near livestock pens. These findings have the potential to inform and improve targeted malaria control and prevention strategies in the region.

Topics: Adolescent; Adult; Agriculture; Animals; Anopheles; Child; Child, Preschool; Conservation of Natural Resources; Female; Humans; Infant; Infant, Newborn; Madagascar; Malaria, Falciparum; Malaria, Vivax; Male; Plasmodium falciparum; Plasmodium vivax; Young Adult

Plasmodium vivax causes the majority of malaria episodes outside Africa, but remains a relatively understudied pathogen. The pathology of P. vivax infection depends critically on the parasite's ability to recognize and invade human erythrocytes. This invasion process involves an interaction between P. vivax Duffy Binding Protein (PvDBP) in merozoites and the Duffy antigen receptor for chemokines (DARC) on the erythrocyte surface. Whole-genome sequencing of clinical isolates recently established that some P. vivax genomes contain two copies of the PvDBP gene. The frequency of this duplication is particularly high in Madagascar, where there is also evidence for P. vivax infection in DARC-negative individuals. The functional significance and global prevalence of this duplication, and whether there are other copy number variations at the PvDBP locus, is unknown.. Using whole-genome sequencing and PCR to study the PvDBP locus in P. vivax clinical isolates, we found that PvDBP duplication is widespread in Cambodia. The boundaries of the Cambodian PvDBP duplication differ from those previously identified in Madagascar, meaning that current molecular assays were unable to detect it. The Cambodian PvDBP duplication did not associate with parasite density or DARC genotype, and ranged in prevalence from 20% to 38% over four annual transmission seasons in Cambodia. This duplication was also present in P. vivax isolates from Brazil and Ethiopia, but not India.. PvDBP duplications are much more widespread and complex than previously thought, and at least two distinct duplications are circulating globally. The same duplication boundaries were identified in parasites from three continents, and were found at high prevalence in human populations where DARC-negativity is essentially absent. It is therefore unlikely that PvDBP duplication is associated with infection of DARC-negative individuals, but functional tests will be required to confirm this hypothesis.

Topics: Adolescent; Antigens, Protozoan; Brazil; Cambodia; Carrier Proteins; Child; Duffy Blood-Group System; Erythrocytes; Ethiopia; Female; Gene Duplication; Humans; India; Madagascar; Malaria, Vivax; Male; Phylogeny; Plasmodium vivax; Protozoan Proteins; Receptors, Cell Surface

Plasmodium vivax is the geographically most widespread human malaria parasite. To analyze patterns of microsatellite diversity and population structure across countries of different transmission intensity, genotyping data from 11 microsatellite markers was either generated or compiled from 841 isolates from four continents collected in 1999-2008. Diversity was highest in South-East Asia (mean allelic richness 10.0-12.8), intermediate in the South Pacific (8.1-9.9) Madagascar and Sudan (7.9-8.4), and lowest in South America and Central Asia (5.5-7.2). A reduced panel of only 3 markers was sufficient to identify approx. 90% of all haplotypes in South Pacific, African and SE-Asian populations, but only 60-80% in Latin American populations, suggesting that typing of 2-6 markers, depending on the level of endemicity, is sufficient for epidemiological studies. Clustering analysis showed distinct clusters in Peru and Brazil, but little sub-structuring was observed within Africa, SE-Asia or the South Pacific. Isolates from Uzbekistan were exceptional, as a near-clonal parasite population was observed that was clearly separated from all other populations (FST>0.2). Outside Central Asia FST values were highest (0.11-0.16) between South American and all other populations, and lowest (0.04-0.07) between populations from South-East Asia and the South Pacific. These comparisons between P. vivax populations from four continents indicated that not only transmission intensity, but also geographical isolation affect diversity and population structure. However, the high effective population size results in slow changes of these parameters. This persistency must be taken into account when assessing the impact of control programs on the genetic structure of parasite populations.

Topics: Africa; Alleles; Americas; Asia; Cluster Analysis; Cohort Studies; Genetic Variation; Genetics, Population; Genotype; Geography; Haplotypes; Humans; Linkage Disequilibrium; Madagascar; Malaria, Vivax; Microsatellite Repeats; Plasmodium vivax

Polymorphic genetic markers and especially microsatellite analysis can be used to investigate multiple aspects of the biology of Plasmodium species. In the current study, we characterized 7 polymorphic microsatellites in a total of 281 Plasmodium vivax isolates to determine the genetic diversity and population structure of P. vivax populations from Sudan, Madagascar, French Guiana, and Armenia. All four parasite populations were highly polymorphic with 3-32 alleles per locus. Mean genetic diversity values was 0.83, 0.79, 0.78 and 0.67 for Madagascar, French Guiana, Sudan, and Armenia, respectively. Significant genetic differentiation between all four populations was observed.

Topics: Alleles; Armenia; French Guiana; Genetic Loci; Genetic Markers; Genetic Variation; Genetics, Population; Heterozygote; Humans; Linkage Disequilibrium; Madagascar; Malaria, Vivax; Microsatellite Repeats; Plasmodium vivax; Spatio-Temporal Analysis; Sudan

Numerous studies have indicated a strong association between amplification of the multidrug resistance-1 gene and in vivo and in vitro mefloquine resistance of Plasmodium falciparum. Although falciparum infection usually is not treated with mefloquine, incorrect diagnosis, high frequency of undetected mixed infections, or relapses of P. vivax infection triggered by P. falciparum infections expose non-P. falciparum parasites to mefloquine. To assess the consequences of such unintentional treatments on P. vivax, we studied variations in number of Pvmdr-1 (PlasmoDB accession no. PVX_080100, NCBI reference sequence NC_009915.1) copies worldwide in 607 samples collected in areas with different histories of mefloquine use from residents and from travelers returning to France. Number of Pvmdr-1 copies correlated with drug use history. Treatment against P. falciparum exerts substantial collateral pressure against sympatric P. vivax, jeopardizing future use of mefloquine against P. vivax. A drug policy is needed that takes into consideration all co-endemic species of malaria parasites.

Topics: Cambodia; Drug Resistance; French Guiana; Gene Expression Regulation; Humans; Madagascar; Malaria, Falciparum; Malaria, Vivax; Mefloquine; Multidrug Resistance-Associated Proteins; Plasmodium falciparum; Plasmodium vivax; Protozoan Proteins; Sudan

In endemic areas, Plasmodium vivax relapses are difficult to distinguish from new infections. Genotyping of patients who experience relapse after returning to a malaria-free area can be used to explore the nature of hypnozoite activation and relapse. This paper describes a person who developed P. vivax malaria for the first time after travelling to Boriziny in the malaria endemic coastal area of Madagascar, then suffered two P. vivax relapses 11 weeks and 21 weeks later despite remaining in Antananarivo in the malaria-free central highlands area. He was treated with the combination artesunate + amodiaquine according to the national malaria policy in Madagascar. Genotyping by PCR-RFLP at pvmsp-3α as well as pvmsp1 heteroduplex tracking assay (HTA) showed the same dominant genotype at each relapse. Multiple recurring minority variants were also detected at each relapse, highlighting the propensity for multiple hypnozoite clones to activate simultaneously to cause relapse.

Topics: Adult; Amodiaquine; Antimalarials; Artemisinins; Drug Combinations; Humans; Madagascar; Malaria, Vivax; Male; Plasmodium vivax; Recurrence; Sporozoites; Young Adult

Plasmodium vivax is the most prevalent human malaria parasite, causing serious public health problems in malaria-endemic countries. Until recently the Duffy-negative blood group phenotype was considered to confer resistance to vivax malaria for most African ethnicities. We and others have reported that P. vivax strains in African countries from Madagascar to Mauritania display capacity to cause clinical vivax malaria in Duffy-negative people. New insights must now explain Duffy-independent P. vivax invasion of human erythrocytes.. Through recent whole genome sequencing we obtained ≥ 70× coverage of the P. vivax genome from five field-isolates, resulting in ≥ 93% of the Sal I reference sequenced at coverage greater than 20×. Combined with sequences from one additional Malagasy field isolate and from five monkey-adapted strains, we describe here identification of DNA sequence rearrangements in the P. vivax genome, including discovery of a duplication of the P. vivax Duffy binding protein (PvDBP) gene. A survey of Malagasy patients infected with P. vivax showed that the PvDBP duplication was present in numerous locations in Madagascar and found in over 50% of infected patients evaluated. Extended geographic surveys showed that the PvDBP duplication was detected frequently in vivax patients living in East Africa and in some residents of non-African P. vivax-endemic countries. Additionally, the PvDBP duplication was observed in travelers seeking treatment of vivax malaria upon returning home. PvDBP duplication prevalence was highest in west-central Madagascar sites where the highest frequencies of P. vivax-infected, Duffy-negative people were reported.. The highly conserved nature of the sequence involved in the PvDBP duplication suggests that it has occurred in a recent evolutionary time frame. These data suggest that PvDBP, a merozoite surface protein involved in red cell adhesion is rapidly evolving, possibly in response to constraints imposed by erythrocyte Duffy negativity in some human populations.

Topics: Antigens, Protozoan; DNA, Protozoan; Gene Duplication; Genome, Protozoan; Humans; Madagascar; Malaria, Vivax; Mauritania; Molecular Sequence Data; Plasmodium vivax; Protozoan Proteins; Receptors, Cell Surface; Sequence Analysis, DNA

An estimated 2.85 billion people live at risk of Plasmodium vivax transmission. In endemic countries vivax malaria causes significant morbidity and its mortality is becoming more widely appreciated, drug-resistant strains are increasing in prevalence, and an increasing number of reports indicate that P. vivax is capable of breaking through the Duffy-negative barrier long considered to confer resistance to blood stage infection. Absence of robust in vitro propagation limits our understanding of fundamental aspects of the parasite's biology, including the determinants of its dormant hypnozoite phase, its virulence and drug susceptibility, and the molecular mechanisms underlying red blood cell invasion.. Here, we report results from whole genome sequencing of five P. vivax isolates obtained from Malagasy and Cambodian patients, and of the monkey-adapted Belem strain. We obtained an average 70-400 X coverage of each genome, resulting in more than 93% of the Sal I reference sequence covered by 20 reads or more. Our study identifies more than 80,000 SNPs distributed throughout the genome which will allow designing association studies and population surveys. Analysis of the genome-wide genetic diversity in P. vivax also reveals considerable allele sharing among isolates from different continents. This observation could be consistent with a high level of gene flow among parasite strains distributed throughout the world.. Our study shows that it is feasible to perform whole genome sequencing of P. vivax field isolates and rigorously characterize the genetic diversity of this parasite. The catalogue of polymorphisms generated here will enable large-scale genotyping studies and contribute to a better understanding of P. vivax traits such as drug resistance or erythrocyte invasion, partially circumventing the lack of laboratory culture that has hampered vivax research for years.

Topics: Adolescent; Cambodia; Child, Preschool; DNA, Protozoan; Female; Genetic Variation; Genome, Protozoan; Humans; Madagascar; Malaria, Vivax; Male; Plasmodium vivax; Sequence Analysis, DNA; Young Adult

Malaria therapy, experimental, and epidemiological studies have shown that erythrocyte Duffy blood group-negative people, largely of African ancestry, are resistant to erythrocyte Plasmodium vivax infection. These findings established a paradigm that the Duffy antigen is required for P. vivax erythrocyte invasion. P. vivax is endemic in Madagascar, where admixture of Duffy-negative and Duffy-positive populations of diverse ethnic backgrounds has occurred over 2 millennia. There, we investigated susceptibility to P. vivax blood-stage infection and disease in association with Duffy blood group polymorphism. Duffy blood group genotyping identified 72% Duffy-negative individuals (FY*B(ES)/*B(ES)) in community surveys conducted at eight sentinel sites. Flow cytometry and adsorption-elution results confirmed the absence of Duffy antigen expression on Duffy-negative erythrocytes. P. vivax PCR positivity was observed in 8.8% (42/476) of asymptomatic Duffy-negative people. Clinical vivax malaria was identified in Duffy-negative subjects with nine P. vivax monoinfections and eight mixed Plasmodium species infections that included P. vivax (4.9 and 4.4% of 183 participants, respectively). Microscopy examination of blood smears confirmed blood-stage development of P. vivax, including gametocytes. Genotyping of polymorphic surface and microsatellite markers suggested that multiple P. vivax strains were infecting Duffy-negative people. In Madagascar, P. vivax has broken through its dependence on the Duffy antigen for establishing human blood-stage infection and disease. Further studies are necessary to identify the parasite and host molecules that enable this Duffy-independent P. vivax invasion of human erythrocytes.

Topics: Adolescent; Asian People; Base Sequence; Black People; Child; Child, Preschool; DNA Primers; Duffy Blood-Group System; Erythrocytes; Female; Genetic Association Studies; Host-Parasite Interactions; Humans; Madagascar; Malaria, Vivax; Male; Molecular Sequence Data; Plasmodium vivax

The Duffy blood group antigen is the portal of entry of the Plasmodiumvivax malaria parasite into human red blood cells and the receptor for a number of CXC and CC chemokines. We review here epidemiological data and evidence derived from therapeutic or experimental human infections associating P. vivax and the Duffy glycoprotein and laboratory studies indicating that P. vivax uses the Duffy antigen as a receptor to invade the red cell. We then review recent field observations indicating that the conclusion of the absolute dependence on the presence of Duffy on the red cell for P. vivax infection and development into the red cell no longer holds true and that in some parts of the world, P. vivax infects and causes disease in Duffy-negative people.

Topics: Africa; Animals; Antimalarials; Blood Group Antigens; Duffy Blood-Group System; Erythrocytes; Gene Deletion; Gene Expression Regulation; Humans; Madagascar; Malaria, Vivax; Papua New Guinea; Phenotype; Plasmodium vivax; Severity of Illness Index; Zoonoses

No data were available concerning Plasmodium vivax resistance to chloroquine (CQ) in Madagascar. We investigated the therapeutic efficacy of CQ in P. vivax malaria, the prevalence of mutations in the pvcrt-o and pvmdr1 genes before treatment, and the association between mutant parasites and the clinical response of the patients to CQ treatment. Clinical isolates were collected at six sentinel sites located in the three epidemiological strata for malaria throughout Madagascar in 2006. Patients were enrolled, treated, and followed up according to the WHO 2001 guidelines for P. vivax infections. Sequencing was used to analyze polymorphisms of the pvcrt-o (exons 1 to 6) and pvmdr1 genes. The treatment failure rate, after adjustment for genotyping, was estimated at 5.1% for the 105 patients included, ranging from zero in the South to 14.8% in the foothills of the Central Highlands. All samples were wild type for pvcrt-o but mutant for the pvmdr1 gene. Ten nonsynonymous mutations were found in the pvmdr1 gene, including five new mutations, four of which were present at low frequencies (1.3% to 7.5%) while the S513R mutation was present at a much higher frequency (96.3%). The other five mutations, including Y976F, had been described before and had frequencies of 97.8% to 100%. Our findings suggest that CQ-resistant P. vivax isolates are present in Madagascar, particularly in the foothills of the Central Highlands. The 976Y pvmdr1 mutation was found not to be useful for monitoring CQ resistance. Further efforts are required to develop suitable tools for monitoring drug resistance in P. vivax malaria.

Topics: Adolescent; Adult; Animals; Antimalarials; Child; Child, Preschool; Chloroquine; Drug Resistance; Female; Humans; Madagascar; Malaria, Vivax; Male; Membrane Transport Proteins; Middle Aged; Molecular Sequence Data; Multidrug Resistance-Associated Proteins; Mutation; Parasitic Sensitivity Tests; Plasmodium vivax; Polymorphism, Genetic; Protozoan Proteins; Sentinel Surveillance; Sequence Analysis, DNA; Treatment Failure; Treatment Outcome; Young Adult

Four of five Plasmodium species infecting humans are present in Madagascar. Plasmodium vivax remains the second most prevalent species, but is understudied. No data is available on its susceptibility to sulphadoxine-pyrimethamine, the drug recommended for intermittent preventive treatment during pregnancy. In this study, the prevalence of P. vivax infection and the polymorphisms in the pvdhfr and pvdhps genes were investigated. The correlation between these polymorphisms and clinical and parasitological responses was also investigated in P. vivax-infected patients.. Plasmodium vivax clinical isolates were collected in eight sentinel sites from the four major epidemiological areas for malaria across Madagascar in 2006/2007. Pvdhfr and pvdhps genes were sequenced for polymorphism analysis. The therapeutic efficacy of SP in P. vivax infections was assessed in Tsiroanomandidy, in the foothill of the central highlands. An intention-to-treat analysis of treatment outcome was carried out.. A total of 159 P. vivax samples were sequenced in the pvdhfr/pvdhps genes. Mutant-types in pvdhfr gene were found in 71% of samples, and in pvdhps gene in 16% of samples. Six non-synonymous mutations were identified in pvdhfr, including two novel mutations at codons 21 and 130. For pvdhps, beside the known mutation at codon 383, a new one was found at codon 422. For the two genes, different combinations were ranged from wild-type to quadruple mutant-type. Among the 16 patients enrolled in the sulphadoxine-pyrimethamine clinical trial (28 days of follow-up) and after adjustment by genotyping, 3 (19%, 95% CI: 5%-43%) of them were classified as treatment failure and were pvdhfr 58R/117N double mutant carriers with or without the pvdhps 383G mutation.. This study highlights (i) that genotyping in the pvdhfr and pvdhps genes remains a useful tool to monitor the emergence and the spread of P. vivax sulphadoxine-pyrimethamine resistant in order to improve the national antimalarial drug policy, (ii) the issue of using sulphadoxine-pyrimethamine as a monotherapy for intermittent preventive treatment of pregnant women or children.

Topics: Adolescent; Adult; Animals; Antimalarials; Child; Child, Preschool; Dihydropteroate Synthase; Drug Combinations; Drug Resistance; Female; Humans; Infant; Madagascar; Malaria, Vivax; Male; Molecular Sequence Data; Mutation; Plasmodium vivax; Polymerase Chain Reaction; Polymorphism, Genetic; Protozoan Proteins; Pyrimethamine; Sequence Analysis, DNA; Sulfadoxine; Tetrahydrofolate Dehydrogenase; Treatment Outcome

Malaria transmission remains poorly documented in areas of low transmission. A study has been carried out over two consecutive years in Analamiranga, a village located at an altitude of 885m on the western edge of the Malagasy highlands, with the aim of generating and updating malariometric indexes for both mosquitoes and schoolchildren. In this village, no vector control measures were performed during the study period nor during previous decades. Mosquitoes were collected monthly when landing on human volunteers and in various resting-places. Blood samples were taken every 3 months from schoolchildren aged 6-12 years and microscopically examined. Of 7,480 mosquitoes collected on human subjects, 5,790 were anophelines. Ten anopheline species were represented and three of these, Anopheles funestus, Anopheles arabiensis and Anopheles mascarensis, accounted for 59.2% of the collection. Of these three species 4,640 were also collected in resting places. The proportion of mosquitoes fed on bovids was high; conversely, the anthropophilic rate (mosquitoes fed on human beings) was especially low: 31%, 7% and 1%, respectively, for A. funestus, A. arabiensis and A. mascarensis. The only confirmed malaria vector was A. funestus with a low sporozoite index (of 6,830 A. funestus, five were positive for Plasmodium falciparum and four for Plasmodium vivax). The annual entomological inoculation rate (number of bites of infected anophelines per adult person) was estimated at 2.49 with low variation over the 2 years. Overall, 909 thick blood smears were tested from blood samples taken from schoolchildren with 30.3% being malaria-positive. The four Plasmodium species infecting human subjects were detected in the following proportions: P. falciparum 78.9%, P. vivax 19.4%, Plasmodium malariae 1.0% and Plasmodium ovale 0.7%. The proportions of children who were infected with any Plasmodium ranged from 10.7% in February to 51.0% in September. Parasitemic children with fever (axillary temperature >37.5 degrees C) accounted for 16.4% of the children sampled. This study demonstrates that there are substantial parasitological consequences of even a relatively low entomological transmission and also recommends including exterior resting-places of mosquitoes in future spraying campaigns in the highlands of Madagascar.

Topics: Animals; Anopheles; Child; Female; Humans; Insect Vectors; Madagascar; Malaria; Malaria, Falciparum; Malaria, Vivax; Population Density; Prevalence; Rural Health; Seasons; Time Factors

Topics: Animals; Anopheles; France; Humans; Insect Vectors; Madagascar; Malaria, Vivax; Male; Middle Aged; Parasitemia; Plasmodium vivax; Travel

Authors record the results of entomological and paludometric studies which were run on July 1994 in Ampanihy (Southern Madagascar), since an epidemic of malaria occurred in that region. Although the risk of epidemic malaria is described in the Southern Madagascar, paludometric indexes found in these surveys are pertinent with a mesoendemic malaria. The entomological survey found. A gambiae complex and A. funestus. Complementary surveys are proposed to improve the knowledge of the epidemiology of malaria in this area.

Topics: Adolescent; Age Distribution; Animals; Anopheles; Child; Child, Preschool; Disease Outbreaks; Female; Humans; Infant; Infant, Newborn; Madagascar; Malaria, Falciparum; Malaria, Vivax; Male; Population Surveillance; Risk Factors; Severity of Illness Index

Plasmodium vivax malaria is prevalent during the rainy season in the central highlands of Madagascar. In April 1991, we investigated the cellular and antibody immune responses of 53 inhabitants of Manarintsoa, a village in this area, to four antigens corresponding to B and T cell epitopes of the P. vivax circumsporozoite (CS) protein. Cellular responses were assessed by lymphocyte proliferation assay as well as by detection of interferon-gamma and interleukin-2 production in vitro. Cell culture was performed with two overlapping synthetic peptides (CSVTCGVGVRVRSRVNA [amino acids 311-326]) and VRVRSRVNAANKKPED [amino acids 319-334]) from the vicinity of the highly conserved region II of the CS protein. In at least one of the three assays, cells from seven subjects showed a positive response to CSVTCGVGVRVRSRVNA, while cells form 14 subjects responded to VRVRSRVNAANKKPED. Antibodies directed against the two recombinant antigens, NS1(81)V20 and rPvCS-2, both of which contain the entire central repeat region of the P. vivax CS protein, plus regions I and II in the case of rPvCS-2, were measured by the Falcon assay screening test-enzyme-linked immunosorbent assay. Eight and nine subjects had antibodies to NS1(81)V20 and rPvCS-2, respectively. The presence of antibody responses to both recombinant antigens was related (P = 0.02, by Fisher's exact test), but was not related to the presence of a cellular response to peptides from vicinity of region II (P > 0.1, by Fisher's exact test).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Amino Acid Sequence; Animals; Antibodies, Protozoan; Antigens, Protozoan; Epitopes; Female; Humans; Immunity, Cellular; Interferon-gamma; Interleukin-2; Lymphocyte Activation; Madagascar; Malaria, Vivax; Male; Middle Aged; Molecular Sequence Data; Plasmodium vivax; Protozoan Proteins; Recombinant Proteins

A study involving 169 schoolchildren (5-14 years old) living in Manarintsoa near Antananarivo (Madagascar, East Africa) was performed during the seasonal malaria transmission period. For the whole population examined, the prevalence of Plasmodium falciparum and the rates of spleen enlargement and of circulating stable antigen (S-Ag) were found to be 60.9%, 71.7%, and 46.8%, respectively. The prevalence of IgG antibody to RESA (ring-infected erythrocyte surface antigen) was 42.7% and that of IgG and IgM antibodies to E-Ag (exoantigens) was 44.9% and 2.9%, respectively. The positive rates for IgG and IgM antibodies to Som-Ag (somatic antigen) were 48.5% and 5.9%, respectively. Concerning S-Ag, no significant relationship was observed for parasitemia, spleen size, age, or IgM antibody responses to exoantigens (E-Ag) or to somatic antigen (Som-Ag). Levels of S-Ag were found to be related to IgG antibodies to E-Ag. Our results suggest that S-Ag at low levels may participate in the mechanisms involved in the development of the IgG antibody responses to E-Ag and to Som-Ag, whereas at a comparative population level, higher quantities of S-Ag down-regulate antibody responses to P. falciparum. The data we obtained were compared with those gathered in another malaria mesoendemic area (Bobo-Dioulasso, Burkina Faso, West Africa), where lower levels of S-Ag were found.

Topics: Adolescent; Age Factors; Animals; Antibody Formation; Antigens, Protozoan; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin G; Immunoglobulin M; Madagascar; Malaria; Malaria, Falciparum; Malaria, Vivax; Plasmodium; Plasmodium falciparum; Plasmodium vivax; Prevalence; Spleen

Topics: Animals; Body Weight; Culicidae; Humans; Madagascar; Malaria; Malaria, Vivax