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1.
Int J Infect Dis ; 143: 107010, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38490637

ABSTRACT

OBJECTIVE: A 15-month longitudinal study was conducted to determine the duration and infectivity of asymptomatic qPCR-detected Plasmodium falciparum and Plasmodium vivax infections in Ethiopia. METHOD: Total parasite and gametocyte kinetics were determined by molecular methods; infectivity to Anopheles arabiensis mosquitoes by repeated membrane feeding assays. Infectivity results were contrasted with passively recruited symptomatic malaria cases. RESULTS: For P. falciparum and P. vivax infections detected at enrolment, median durations of infection were 37 days (95% confidence interval [CI], 15-93) and 60 days (95% CI, 18-213), respectively. P. falciparum and P. vivax parasite densities declined over the course of infections. From 47 feeding assays on 22 asymptomatic P. falciparum infections, 6.4% (3/47) were infectious and these infected 1.8% (29/1579) of mosquitoes. No transmission was observed in feeding assays on asymptomatic P. vivax mono-infections (0/56); one mixed-species infection was highly infectious. Among the symptomatic cases, 4.3% (2/47) of P. falciparum and 73.3% (53/86) of P. vivax patients were infectious to mosquitoes. CONCLUSION: The majority of asymptomatic infections were of short duration and low parasite density. Only a minority of asymptomatic individuals were infectious to mosquitoes. This contrasts with earlier findings and is plausibly due to the low parasite densities in this population.


Subject(s)
Anopheles , Malaria, Falciparum , Malaria, Vivax , Plasmodium falciparum , Plasmodium vivax , Ethiopia/epidemiology , Malaria, Vivax/transmission , Malaria, Vivax/epidemiology , Malaria, Vivax/parasitology , Humans , Longitudinal Studies , Malaria, Falciparum/transmission , Malaria, Falciparum/epidemiology , Malaria, Falciparum/parasitology , Animals , Plasmodium vivax/isolation & purification , Plasmodium vivax/physiology , Plasmodium falciparum/isolation & purification , Anopheles/parasitology , Male , Female , Adult , Adolescent , Child , Young Adult , Child, Preschool , Asymptomatic Infections/epidemiology , Mosquito Vectors/parasitology , Middle Aged
2.
Nat Med ; 29(12): 3203-3211, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37884028

ABSTRACT

Anopheles stephensi, an Asian malaria vector, continues to expand across Africa. The vector is now firmly established in urban settings in the Horn of Africa. Its presence in areas where malaria resurged suggested a possible role in causing malaria outbreaks. Here, using a prospective case-control design, we investigated the role of An. stephensi in transmission following a malaria outbreak in Dire Dawa, Ethiopia in April-July 2022. Screening contacts of patients with malaria and febrile controls revealed spatial clustering of Plasmodium falciparum infections around patients with malaria in strong association with the presence of An. stephensi in the household vicinity. Plasmodium sporozoites were detected in these mosquitoes. This outbreak involved clonal propagation of parasites with molecular signatures of artemisinin and diagnostic resistance. To our knowledge, this study provides the strongest evidence so far for a role of An. stephensi in driving an urban malaria outbreak in Africa, highlighting the major public health threat posed by this fast-spreading mosquito.


Subject(s)
Anopheles , Malaria, Falciparum , Malaria , Animals , Humans , Malaria/epidemiology , Malaria/parasitology , Anopheles/parasitology , Mosquito Vectors/parasitology , Malaria, Falciparum/diagnosis , Malaria, Falciparum/epidemiology , Ethiopia/epidemiology
3.
Front Cell Infect Microbiol ; 12: 1106369, 2022.
Article in English | MEDLINE | ID: mdl-36726645

ABSTRACT

Naturally acquired antibodies may reduce the transmission of Plasmodium gametocytes to mosquitoes. Here, we investigated associations between antibody prevalence and P. vivax infectivity to mosquitoes. A total of 368 microscopy confirmed P. vivax symptomatic patients were passively recruited from health centers in Ethiopia and supplemented with 56 observations from asymptomatic P. vivax parasite carriers. Direct membrane feeding assays (DMFA) were performed to assess mosquito infectivity; for selected feeds these experiments were also performed after replacing autologous plasma with malaria naïve control serum (n=61). The prevalence of antibodies against 6 sexual stage antigens (Pvs47, Pvs48/45, Pvs230, PvsHAP2, Pvs25 and PvCelTOS) and an array of asexual antigens was determined by ELISA and multiplexed bead-based assays. Gametocyte (ρ< 0.42; p = 0.0001) and parasite (ρ = 0.21; p = 0.0001) densities were positively associated with mosquito infection rates. Antibodies against Pvs47, Pvs230 and Pvs25 were associated with 23 and 34% reductions in mosquito infection rates (p<0.0001), respectively. Individuals who showed evidence of transmission blockade in serum-replacement DMFAs (n=8) were significantly more likely to have PvsHAP2 or Pvs47 antibodies. Further studies may demonstrate causality for the observed associations, improve our understanding of the natural transmission of P. vivax and support vaccine development.


Subject(s)
Anopheles , Malaria, Vivax , Malaria , Animals , Humans , Plasmodium vivax , Anopheles/parasitology , Malaria, Vivax/prevention & control , Antibodies, Protozoan , Plasmodium falciparum
4.
Emerg Infect Dis ; 27(2): 603-607, 2021 02.
Article in English | MEDLINE | ID: mdl-33496217

ABSTRACT

Anopheles stephensi mosquitoes, efficient vectors in parts of Asia and Africa, were found in 75.3% of water sources surveyed and contributed to 80.9% of wild-caught Anopheles mosquitoes in Awash Sebat Kilo, Ethiopia. High susceptibility of these mosquitoes to Plasmodium falciparum and vivax infection presents a challenge for malaria control in the Horn of Africa.


Subject(s)
Anopheles , Plasmodium vivax , Animals , Asia , Ethiopia , Mosquito Vectors , Plasmodium falciparum
5.
Malar J ; 20(1): 59, 2021 Jan 22.
Article in English | MEDLINE | ID: mdl-33482841

ABSTRACT

BACKGROUND: As countries move to malaria elimination, detecting and targeting asymptomatic malaria infections might be needed. Here, the epidemiology and detectability of asymptomatic Plasmodium falciparum and Plasmodium vivax infections were investigated in different transmission settings in Ethiopia. METHOD: A total of 1093 dried blood spot (DBS) samples were collected from afebrile and apparently healthy individuals across ten study sites in Ethiopia from 2016 to 2020. Of these, 862 were from community and 231 from school based cross-sectional surveys. Malaria infection status was determined by microscopy or rapid diagnostics tests (RDT) and 18S rRNA-based nested PCR (nPCR). The annual parasite index (API) was used to classify endemicity as low (API > 0 and < 5), moderate (API ≥ 5 and < 100) and high transmission (API ≥ 100) and detectability of infections was assessed in these settings. RESULTS: In community surveys, the overall prevalence of asymptomatic Plasmodium infections by microscopy/RDT, nPCR and all methods combined was 12.2% (105/860), 21.6% (183/846) and 24.1% (208/862), respectively. The proportion of nPCR positive infections that was detectable by microscopy/RDT was 48.7% (73/150) for P. falciparum and 4.6% (2/44) for P. vivax. Compared to low transmission settings, the likelihood of detecting infections by microscopy/RDT was increased in moderate (Adjusted odds ratio [AOR]: 3.4; 95% confidence interval [95% CI] 1.6-7.2, P = 0.002) and high endemic settings (AOR = 5.1; 95% CI 2.6-9.9, P < 0.001). After adjustment for site and correlation between observations from the same survey, the likelihood of detecting asymptomatic infections by microscopy/RDT (AOR per year increase = 0.95, 95% CI 0.9-1.0, P = 0.013) declined with age. CONCLUSIONS: Conventional diagnostics missed nearly half of the asymptomatic Plasmodium reservoir detected by nPCR. The detectability of infections was particularly low in older age groups and low transmission settings. These findings highlight the need for sensitive diagnostic tools to detect the entire parasite reservoir and potential infection transmitters.


Subject(s)
Asymptomatic Infections/epidemiology , Malaria, Falciparum/diagnosis , Malaria, Falciparum/epidemiology , Malaria, Vivax/diagnosis , Malaria, Vivax/epidemiology , Adolescent , Adult , Child , Cross-Sectional Studies , Dried Blood Spot Testing , Ethiopia/epidemiology , Female , Humans , Malaria, Falciparum/transmission , Malaria, Vivax/transmission , Male , Microscopy/methods , Middle Aged , Plasmodium falciparum/genetics , Plasmodium vivax/genetics , Polymerase Chain Reaction , Prevalence , RNA, Ribosomal, 18S , Young Adult
6.
Sci Rep ; 10(1): 19975, 2020 11 17.
Article in English | MEDLINE | ID: mdl-33203956

ABSTRACT

The distribution of malaria infections is heterogeneous in space and time, especially in low transmission settings. Understanding this clustering may allow identification and targeting of pockets of transmission. In Adama district, Ethiopia, Plasmodium falciparum and P. vivax malaria patients and controls were examined, together with household members and immediate neighbors. Rapid diagnostic test and quantitative PCR (qPCR) were used for the detection of infections that were genetically characterized by a panel of microsatellite loci for P. falciparum (26) and P. vivax (11), respectively. Individuals living in households of clinical P. falciparum patients were more likely to have qPCR detected P. falciparum infections (22.0%, 9/41) compared to individuals in control households (8.7%, 37/426; odds ratio, 2.9; 95% confidence interval, 1.3-6.4; P = .007). Genetically related P. falciparum, but not P. vivax infections showed strong clustering within households. Genotyping revealed a marked temporal cluster of P. falciparum infections, almost exclusively comprised of clinical cases. These findings uncover previously unappreciated transmission dynamics and support a rational approach to reactive case detection strategies for P. falciparum in Ethiopia.


Subject(s)
Malaria, Falciparum/parasitology , Malaria, Vivax/parasitology , Plasmodium falciparum/genetics , Plasmodium vivax/genetics , Cluster Analysis , Ethiopia , Family Characteristics , Genotype , Humans , Polymerase Chain Reaction/methods
7.
Parasit Vectors ; 13(1): 120, 2020 Mar 06.
Article in English | MEDLINE | ID: mdl-32143713

ABSTRACT

BACKGROUND: Mosquito-feeding assays that assess transmission of Plasmodium from man-to-mosquito typically use laboratory mosquito colonies. The microbiome and genetic background of local mosquitoes may be different and influence Plasmodium transmission efficiency. In order to interpret transmission studies to the local epidemiology, it is therefore crucial to understand the relationship between infectivity in laboratory-adapted and local mosquitoes. METHODS: We assessed infectivity of Plasmodium vivax-infected patients from Adama, Ethiopia, using laboratory-adapted (colony) and wild-caught (wild) mosquitoes raised from larval collections in paired feeding experiments. Feeding assays used 4-6 day-old female Anopheles arabiensis mosquitoes after starvation for 12 h (colony) and 18 h (wild). Oocyst development was assessed microscopically 7 days post-feeding. Wild mosquitoes were identified morphologically and confirmed by genotyping. Asexual parasites and gametocytes were quantified in donor blood by microscopy. RESULTS: In 36 paired experiments (25 P. vivax infections and 11 co-infections with P. falciparum), feeding efficiency was higher in colony (median: 62.5%; interquartile range, IQR: 47.0-79.0%) compared to wild mosquitoes (median: 27.8%; IQR: 17.0-38.0%; Z = 5.02; P < 0.001). Plasmodium vivax from infectious individuals (51.6%, 16/31) infected a median of 55.0% (IQR: 6.7-85.7%; range: 5.5-96.7%; n = 14) of the colony and 52.7% (IQR: 20.0-80.0%; range: 3.2-95.0%; n = 14) of the wild mosquitoes. A strong association (ρ(16) = 0.819; P < 0.001) was observed between the proportion of infected wild and colony mosquitoes. A positive association was detected between microscopically detected gametocytes and the proportion of infected colony (ρ(31) = 0.452; P = 0.011) and wild (ρ(31) = 0.386; P = 0.032) mosquitoes. CONCLUSIONS: Infectivity assessments with colony and wild mosquitoes yielded similar infection results. This finding supports the use of colony mosquitoes for assessments of the infectious reservoir for malaria in this setting whilst acknowledging the importance of mosquito factors influencing sporogonic development of Plasmodium parasites.


Subject(s)
Anopheles/physiology , Anopheles/parasitology , Laboratories , Malaria, Vivax/parasitology , Mosquito Vectors/physiology , Mosquito Vectors/parasitology , Plasmodium vivax/physiology , Animals , Ethiopia , Feeding Behavior/physiology , Female , Host-Parasite Interactions , Humans , Larva , Malaria/transmission , Oocysts/growth & development , Plasmodium vivax/genetics
8.
Am J Trop Med Hyg ; 102(1): 117-120, 2020 01.
Article in English | MEDLINE | ID: mdl-31733053

ABSTRACT

Ethiopia aims to diagnose and treat all clinical malaria within 24 hours of fever onset in its stride to eliminate the disease by 2030. Microscopy remains to be the mainstay for diagnosis at the health center and hospital level. Continuous evaluation and performance upgrading of malaria microscopists is one of the cornerstones in this effort. We assessed the performance of malaria microscopists compared with reference readers in diagnosing, identifying the species, and quantifying parasitemia. A total of 174 microscopists were enrolled from health facilities located in 86 districts in Oromia region (Ethiopia) from January 2017 to June 2018. Panel slides with known Plasmodium species, diagnostic blood stage, and parasite density were prepared by the reference readers. Sociodemographics, education, in-service training, and routine practice of participants were captured. Sensitivity, specificity, percent agreement, and kappa score were calculated. An overall low performance was observed that could threaten the malaria diagnostic service. Of all the slides distributed (1,218), only 17.0% of the positive and 30.0% of the negative slides were correctly identified and 22.4% were correctly quantified. Compared with the reference readers, participants had lower competence in diagnosing (74.3% agreement and kappa 0.45) and identifying the species (71.2% agreement and kappa 0.40). Two-fifths of the participants were graded as "in training" with respect to identifying the species (41.0%) and the diagnostic stages (40.0%). An in-service training/retraining and supportive supervision are needed to raise and maintain the competence of microscopists in settings with a recent decline in malaria transmission and aiming for ultimate elimination of the disease.


Subject(s)
Laboratory Personnel/standards , Malaria, Falciparum/diagnosis , Malaria, Vivax/diagnosis , Microscopy/standards , Adult , Clinical Laboratory Techniques/standards , Diagnostic Errors , Ethiopia/epidemiology , Female , Humans , Malaria, Falciparum/epidemiology , Malaria, Vivax/epidemiology , Male , Sensitivity and Specificity
9.
Article in English | MEDLINE | ID: mdl-31539706

ABSTRACT

Plasmodium falciparum and P. vivax co-exist at different endemicity levels across Ethiopia. For over two decades Artemether-Lumefantrine (AL) is the first line treatment for uncomplicated P. falciparum, while chloroquine (CQ) is still used to treat P. vivax. It is currently unclear whether a shift from CQ to AL for P. falciparum treatment has implications for AL efficacy and results in a reversal of mutations in genes associated to CQ resistance, given the high co-endemicity of the two species and the continued availability of CQ for the treatment of P. vivax. This study thus assessed the prevalence of Pfcrt-K76T and Pfmdr1-N86Y point mutations in P. falciparum. 18S RNA gene based nested PCR confirmed P. falciparum samples (N = 183) collected through community and health facility targeted cross-sectional surveys from settings with varying P. vivax and P. falciparum endemicity were used. The proportion of Plasmodium infections that were P. vivax was 62.2% in Adama, 41.4% in Babile, 30.0% in Benishangul-Gumuz to 6.9% in Gambella. The Pfcrt-76T mutant haplotype was observed more from samples with higher endemicity of P. vivax as being 98.4% (61/62), 100% (31/31), 65.2% (15/23) and 41.5% (22/53) in samples from Adama, Babile, Benishangul-Gumuz and Gambella, respectively. However, a relatively higher proportion of Pfmdr1-N86 allele (77.3-100%) were maintained in all sites. The observed high level of the mutant Pfcrt-76T allele in P. vivax co-endemic sites might require that utilization of CQ needs to be re-evaluated in settings co-endemic for the two species. A country-wide assessment is recommended to clarify the implication of the observed level of variation in drug resistance markers on the efficacy of AL-based treatment against uncomplicated P. falciparum malaria.


Subject(s)
Membrane Transport Proteins/genetics , Multidrug Resistance-Associated Proteins/genetics , Plasmodium falciparum/genetics , Plasmodium vivax/genetics , Protozoan Proteins/genetics , Adolescent , Adult , Alleles , Antimalarials/pharmacology , Antimalarials/therapeutic use , Artemether, Lumefantrine Drug Combination/pharmacology , Artemether, Lumefantrine Drug Combination/therapeutic use , Child , Chloroquine/pharmacology , Chloroquine/therapeutic use , Drug Resistance , Endemic Diseases , Ethiopia/epidemiology , Female , Haplotypes , Humans , Malaria, Falciparum/complications , Malaria, Falciparum/drug therapy , Malaria, Falciparum/epidemiology , Malaria, Falciparum/parasitology , Malaria, Vivax/complications , Malaria, Vivax/drug therapy , Malaria, Vivax/epidemiology , Malaria, Vivax/parasitology , Male , Plasmodium falciparum/classification , Plasmodium falciparum/physiology , Plasmodium vivax/classification , Plasmodium vivax/physiology , Point Mutation , Polymorphism, Genetic , Prevalence , Young Adult
10.
Trans R Soc Trop Med Hyg ; 113(6): 305-311, 2019 06 01.
Article in English | MEDLINE | ID: mdl-30927007

ABSTRACT

BACKGROUND: Following successful malaria control during the last decade, Ethiopia instituted a stepwise malaria elimination strategy in selected low-transmission areas. METHODS: Cross-sectional surveys were conducted in Babile district, Oromia, Ethiopia from July to November 2017 to evaluate malaria infection status using microscopy and nested polymerase chain reaction (nPCR) and serological markers of exposure targeting Plasmodium falciparum and Plasmodium vivax apical membrane antigen-1 (AMA-1). RESULTS: Parasite prevalence was 1.2% (14/1135) and 5.1% (58/1143) for P. falciparum and 0.4% (5/1135) and 3.6% (41/1143) for P. vivax by microscopy and nPCR, respectively. Antibody prevalence was associated with current infection by nPCR for both P. falciparum (p<0.001) and P. vivax (p=0.014) and showed an age-dependent increase (p<0.001, for both species). Seroconversion curves indicated a decline in malaria exposure 15 y prior to sampling for P. falciparum and 11.5 y prior to sampling for P. vivax, broadly following malaria incidence data from district health offices, with higher antibody titres in adults than children for both species. CONCLUSIONS: Malaria transmission declined substantially in the region with continuing heterogeneous but measurable local transmission, arguing in favour of continued and tailored control efforts to accelerate the progress towards elimination efforts.


Subject(s)
Malaria, Falciparum/epidemiology , Malaria, Vivax/epidemiology , Adolescent , Adult , Child , Child, Preschool , Cross-Sectional Studies , Ethiopia/epidemiology , Female , Humans , Malaria, Falciparum/transmission , Malaria, Vivax/transmission , Male , Middle Aged , Plasmodium falciparum/isolation & purification , Plasmodium vivax/isolation & purification , Prevalence , Seroepidemiologic Studies , Young Adult
11.
Malar J ; 17(1): 281, 2018 Aug 02.
Article in English | MEDLINE | ID: mdl-30071859

ABSTRACT

BACKGROUND: 8-Aminoquinolines such as primaquine clear mature Plasmodium falciparum gametocytes that are responsible for transmission from human to mosquitoes and bring radical cure in Plasmodium vivax by clearing dormant liver stages. Deployment of primaquine is thus of relevance for malaria elimination efforts but challenged by the widespread prevalence of glucose-6-phosphate dehydrogenase deficiency (G6PDd) in endemic countries since primaquine in G6PDd individuals may lead to acute haemolysis. In this study, the prevalence of G6PDd was investigated in different settings in Ethiopia using phenotyping and genotyping approaches. METHODS: Community and school based cross-sectional surveys were conducted from October to December 2016 in four administrative regions (Gambela, Benishangul Gumuz, Oromia, and Amhara) in Ethiopia. Finger prick blood samples were collected for G6PD enzyme activity using the CareStart™ G6PD screening test and genotyping of 36 selected single nucleotide polymorphisms (SNPs) located in the G6PD gene and its flanking regions. RESULTS: Overall, the prevalence of phenotypic G6PDd was 1.4% (22/1609). For the first time in the Ethiopian population, the African variant (A-) was detected in 3.5% (7/199) of the limited set of genotyped samples, which were all phenotypically normal. Interestingly, all of these individuals had a variation at the rs2515904 locus. Strong geographical variation was observed for both phenotypic and genotypic G6PDd; three-quarters of the phenotypically G6PDd individuals were detected in Gambela. CONCLUSION: A very low prevalence of G6PDd was detected in the present study populations. The presence of the A- variant alongside other G6PD mutants and the patchy distribution of G6PDd indicate that larger studies specifically designed to unravel the distribution of G6PDd at small geographical scale may be needed to tailor malaria elimination efforts in Ethiopia to the local context.


Subject(s)
Glucosephosphate Dehydrogenase Deficiency/epidemiology , Polymorphism, Single Nucleotide , Adolescent , Adult , Child , Cross-Sectional Studies , Ethiopia/epidemiology , Female , Genotype , Glucosephosphate Dehydrogenase Deficiency/genetics , Glucosephosphate Dehydrogenase Deficiency/parasitology , Humans , Male , Phenotype , Prevalence , Young Adult
12.
Clin Infect Dis ; 66(12): 1883-1891, 2018 06 01.
Article in English | MEDLINE | ID: mdl-29304258

ABSTRACT

Background: The majority of Plasmodium vivax and Plasmodium falciparum infections in low-endemic settings are asymptomatic. The relative contribution to the infectious reservoir of these infections compared to clinical malaria cases is currently unknown. Methods: We assessed infectivity of passively recruited symptomatic malaria patients (n = 41) and community-recruited asymptomatic individuals with microscopy-detected (n = 41) and polymerase chain reaction (PCR)-detected infections (n = 82) using membrane feeding assays with Anopheles arabiensis mosquitoes in Adama, Ethiopia. Malaria incidence and prevalence data were used to estimate the contributions of these populations to the infectious reservoir. Results: Overall, 34.9% (29/83) of P. vivax- and 15.1% (8/53) P. falciparum-infected individuals infected ≥1 mosquitoes. Mosquito infection rates were strongly correlated with asexual parasite density for P. vivax (ρ = 0.63; P < .001) but not for P. falciparum (ρ = 0.06; P = .770). Plasmodium vivax symptomatic infections were more infectious to mosquitoes (infecting 46.5% of mosquitoes, 307/660) compared to asymptomatic microscopy-detected (infecting 12.0% of mosquitoes, 80/667; P = .005) and PCR-detected infections (infecting 0.8% of mosquitoes, 6/744; P < .001). Adjusting for population prevalence, symptomatic, asymptomatic microscopy-detected, and PCR-detected infections were responsible for 8.0%, 76.2%, and 15.8% of the infectious reservoir for P. vivax, respectively. For P. falciparum, mosquito infections were sparser and also predominantly from asymptomatic infections. Conclusions: In this low-endemic setting aiming for malaria elimination, asymptomatic infections were highly prevalent and responsible for the majority of onward mosquito infections. The early identification and treatment of asymptomatic infections might accelerate elimination efforts.


Subject(s)
Anopheles/parasitology , Asymptomatic Infections/epidemiology , Disease Reservoirs/parasitology , Malaria, Falciparum/epidemiology , Malaria, Vivax/epidemiology , Adolescent , Adult , Animals , Child , Child, Preschool , Endemic Diseases/statistics & numerical data , Ethiopia/epidemiology , Female , Humans , Malaria, Falciparum/transmission , Malaria, Vivax/transmission , Male , Plasmodium falciparum/genetics , Plasmodium falciparum/isolation & purification , Plasmodium vivax/genetics , Plasmodium vivax/isolation & purification , Polymerase Chain Reaction , Prevalence , Young Adult
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