- Open Access
Asymptomatic only at first sight: malaria infection among schoolchildren in highland Rwanda
- Kevin C. Sifft1,
- Dominik Geus1,
- Caritas Mukampunga2,
- Jean Claude Mugisha2,
- Felix Habarugira2,
- Kira Fraundorfer3,
- Claude Bayingana2,
- Jules Ndoli2,
- Irenee Umulisa4,
- Corine Karema4, 5, 6,
- George von Samson-Himmelstjerna3,
- Toni Aebischer7,
- Peter Martus8,
- Augustin Sendegeya2,
- Jean Bosco Gahutu2 and
- Frank P. Mockenhaupt1Email author
© The Author(s) 2016
Received: 7 September 2016
Accepted: 7 November 2016
Published: 14 November 2016
Plasmodium infection and malaria in school children are increasingly recognized as a relevant public health problem, but data on actual prevalence and health consequences are insufficient. The present study from highland southern Rwanda aimed at estimating infection prevalence among children attending school, at identifying associated factors and at assessing the clinical consequences of these infections.
In a survey including 12 schools in the Huye district of Rwanda, 1089 children aged 6–10 years were clinically and anthropometrically examined, malaria parasites were diagnosed by microscopy and PCR, haemoglobin concentrations were measured, and socio-economic and behavioural parameters as well as medical histories were obtained.
Upon examination, the vast majority of children was asymptomatic (fever 2.7%). Plasmodium infection was detected in 22.4% (Plasmodium falciparum, 18.8%); 41% of these were submicroscopic. Independent predictors of infection included low altitude, higher age, preceding antimalarial treatment, and absence of electricity or a bicycle in the household. Plasmodium infection was associated with anaemia (mean haemoglobin difference of −1.2 g/dL; 95% CI, −0.8 to −1.5 g/dL), fever, underweight, clinically assessed malnutrition and histories of fever, tiredness, weakness, poor appetite, abdominal pain, and vomiting. With the exception of underweight, these conditions were also increased at submicroscopic infection.
Malaria infection is frequent among children attending school in southern highland Rwanda. Although seemingly asymptomatic in the vast majority of cases, infection is associated with a number of non-specific symptoms in the children´s histories, in addition to the impact on anaemia. This argues for improved malaria surveillance and control activities among school children.
Plasmodium infection can cause a wide variety of illnesses, ranging from asymptomatic infection over uncomplicated malaria to severe and complicated disease [1–4]. Owing to the enormous morbidity and mortality of clinical malaria in young children, the epidemiology and consequences of asymptomatic infection in different settings and age-groups has received comparatively little attention. Increasingly, however, the underrated impact of asymptomatic and/or submicroscopic infections is now being recognized [2, 3, 5–10].
Partial immunity against malaria develops during childhood in endemic regions as a result of repeated exposure. Increasing age has consequently been associated with low and submicroscopic parasite density as well as with asymptomatic infection, particularly among school-age children [5, 11–14]. Most of these infections remain undiagnosed and untreated [14, 15], and—in the case of schoolchildren—are not subject of targeted interventions. However, the significance of these asymptomatic Plasmodium infections is likely underestimated as they contribute to inflammatory responses, anaemia, cognitive impairment and delayed physical development [5, 9, 10, 15–20]. Moreover, submicroscopic infections tend to persist for longer than symptomatic ones [13, 21], thereby constituting a sizeable transmission reservoir [2, 3, 14]. This has particular importance in areas of low endemicity considering the increase of submicroscopic (and frequently asymptomatic and untreated) infections with declining malaria transmission [2, 3, 11, 21].
In Rwanda, comprehensive malaria control measures since 2005 have been paralleled by 50% or higher declines in malaria morbidity and mortality . In 2010, in the southern highland Huye district, PCR-assessed Plasmodium prevalence among rural children under 5 years of age was 16%, the majority of infections being asymptomatic . This area represents a hypo- to meso-endemic setting as seen in many other parts of the country . No data, however, is available as to the epidemiological characteristics and effects of seemingly asymptomatic and/or submicroscopic Plasmodium infections among school-age children living in the highlands of Rwanda. Such knowledge, nevertheless, forms the prerequisites for the design of targeted interventions, the reduction of transmission reservoirs, and, ultimately, elimination.
In 2014, children attending primary schools in the largely rural Huye district of highland southern Rwanda were recruited for a study on anthelminthic treatment efficacy. Based on this study population, the present analysis aimed at estimating the prevalence of Plasmodium infection including submicroscopic ones, at identifying socio-demographic and -economic as well as behavioural factors associated with infection, and at assessing the clinical manifestation of these infections with particular focus on anaemia and non-specific symptoms.
Study design, setting and population
This observational, cross-sectional study was conducted in October 2014 in the Huye district, Southern Province, Rwanda. Huye district (population 330,000) is located on Rwanda’s central plateau, which is characterized by an average altitude of 1600–1800 m, a mean temperature of 19 °C and a yearly rainfall of approximately 1200 mm with two rainy seasons (October–December; March–May). Malaria transmission in the area is perennial but low with peaks during the rainy seasons. Plasmodium falciparum is the predominant species [12, 18, 23].
Examinations and laboratory procedures
Alongside routine deworming activities, all participating schools were visited within one week in October 2014. Attending children were clinically examined, and axillary temperature was measured. A brief medical history was obtained from the children, and results were combined with the history as stated by the parents upon interviewing (see below). Age, sex, weight, height, mid-upper arm circumference (MUAC), and fever (axillary temperature ≥37.5 °C) were documented. Weight-for-age z (WAZ) scores and height-for-age z (HAZ) scores were calculated using AnthroPlus . Underweight and stunting were defined as a WAZ or HAZ score < −2 SD, respectively. In addition, clinically assessed malnutrition as determined by the attending physician upon physical examination was documented. Peripheral venous blood was collected into EDTA. Haemoglobin (Hb) concentrations were measured using a HemoCue photometer (Ängelholm, Sweden), and anaemia was defined as a haemoglobin concentration <12 g/dL according to age-related and altitude-adjusted cut-off values . On-site, P. falciparum and non-falciparum malaria parasites were detected by immunochromatographic dipstick tests (SD BIOLINE Malaria Ag P.f/Pan, Standard Diagnostics Inc, Gyeonggi-do, Korea); children found positive were treated with artemether–lumefantrine. In case of further illness upon clinical examination, treatment was provided according to Rwandan treatment guidelines, if needed. Blood samples were transported on wet ice to the central laboratory, University Teaching Hospital of Butare (UTBH). Malaria parasites were counted on Giemsa-stained thick blood films per 200 white blood cells (WBCs), and parasite density was calculated assuming a mean WBC count of 8000/µL. A slide was declared negative only after having counted 500 WBCs. Malaria was defined as any microscopically visible parasitaemia plus fever or a history of fever within the preceding 48 h. DNA was extracted from stabilized full blood aliquots (AS1 & Qiamp blood kit; Qiagen, Germany), and Plasmodium species and submicroscopic infections were identified by semi-nested multiplex polymerase chain reaction (PCR) assays . Plasmodium spp. infection was defined as any parasitaemia detected either by microscopy or PCR. Asymptomatic infection herein is defined as the presence of Plasmodium spp. in the absence of fever upon examination. This does not exclude the presence of signs and symptoms in the history of the children.
Socio-economic status and household visits
Trained health workers visited the households of the recruited children and conducted interviews with parents or guardians using a standardized questionnaire. The questionnaire in the local Kinyarwanda language was largely based on precursors used in previous studies in the region . Data collected included information on paternal education and occupation, household characteristics and assets, ability to pay school and insurance fees, ownership and type of livestock, sanitary facilities, water supply and mosquito net ownership and use as well as illnesses or symptoms the child suffered within the preceding 2 weeks, treatment-seeking, and current or past drug use including anti-malarials.
Data from questionnaires, clinical examination, and laboratory results were double-entered into electronic databases and cross-checked. SPSS (SPSS Statistics Version 23.0, IBM Corp., Armonk, NY, USA) was used for statistical data analysis. Geometric mean parasite densities (GMPDs) and 95% confidence intervals (95% CIs) were calculated. Continuous variables were compared between groups by Student’s t test, Mann–Whitney or Kruskal–Wallis tests, and proportions by χ2 test or Fisher’s exact test. Odds ratios (ORs) and 95% CIs were computed. Evaluation of determinants of Plasmodium infection and of clinical symptoms was performed by simple and multiple logistic regression analysis. Analyses were adjusted for intracluster correlations (clusters = schools) by use of generalized estimating equations (GEE, exchangeable correlation structure for subjects within identical villages). Multivariate analysis of factors associated with Plasmodium infection was done for the complete sample using multiple imputation for missing data. Stepwise backward selection was performed, and final models included those factors that retained statistical significance. A P value <0.05 was considered statistically significant.
Characteristics of 1089 Rwandan schoolchildren
Female (%, n/n)
Age (years; median, range)
Weight (kg; median, range)a
Height (cm; median, range)b
Weight-for-age Z-score (WAZ; median, range)a
Underweight (WAZ < 2; %, n/n)
Height-for-age Z-score (HAZ; median, range)b
Stunting (HAZ < 2; %, n/n)
MUAC (cm; mean ± SD)a
16.2 ± 1.3
Clinically assessed malnutrition (%, n/n)
Axillary temperature (°C; mean ± SD)
36.5 ± 0.54
Fever (%, n/n)
Hb (g/dL; median, range)a
Anaemia (%, n/n)
History in preceding 2 weeks (%, n/n)
Plasmodium infection (%, n/n)
Plasmodium spp. PCR positive
P. falciparum, PCR positive
P. ovale, PCR positive
P. malariae, PCR positive
Plasmodium spp., microscopy positivec
Geometric mean parasite density (/µL; 95% CI)
Selected socio-economic characteristics of 1019 Rwandan schoolchildren
Maternal education (%, n/n)
Secondary or higher; else
Paternal education (%, n/n)
Secondary or higher; else
Mother’s occupation farmer or labourer
Father’s occupation farmer or labourer
No. of people in household (median, range)a
No. of siblings (median, range)a
No. of rooms in household (median, range)b
Farmland ownership (%, n/n)
Household assets (%, n/n)
Ability to treat water
Floor material soil (%, n/n)
Unable to pay school fee (%, n/n)
Unable to pay health insurance (%, n/n)
Slept under a bed net last night (%, n/n)
Attended health facility within last two weeks (%, n/n)
Received anti-malarial treatment within last month (%, n/n)
Time to reach next health centre less than 30 min (%, n/n)
Questionnaire data on SES were available for 1019 participants and reflected a generally poor, rural population (Table 2). Parental primary education predominated but 16% had no formal education at all. The vast majority of children (83%) had slept under a bed net the night before the survey but this proportion declined with age to 78% (168/215) among children aged 10 years (P = 0.04 for comparison across years of age).
Infection with Plasmodium spp. as detected by PCR was present in more than a fifth of all children and was largely due to P. falciparum (Table 1); 41% of these infections were submicroscopic, i.e., below the threshold of microscopy but detected by PCR.
Factors associated with Plasmodium infection
Univariate and multivariate analysis of factors associated with Plasmodium infection (PCR) among Rwandan schoolchildren
Proportion infected (%, n)
Stunting (HAZ < 2 SD)
Residence altitude (m)
No. of rooms in household
Electricity in household
Radio in household
Bicycle in household
TV set in household
Mobile phone in household
Ability to pay school fees
Ability to pay health insurance
Child slept under bed net last night
Child attended health facility within preceding 2 weeks
Child received any malaria treatment within preceding month
Time to reach next health centre (min)
Concurrent infection with soil-transmitted helminths (virtually all Ascaris lumbricoides, detected by PCR ) was not associated with Plasmodium infection: 22.5% (72/320) and 20.7% (124/600) of A. lumbricoides infected and non-infected children, respectively, harboured Plasmodium parasites (P = 0.52).
Clinical manifestations of Plasmodium infection
Clinical manifestations of Plasmodium spp. infection among 1071 Rwandan schoolchildren
Infected (PCR positive)
Hb (g/dL; median, range)
Current fever (%)
Ax. Temperature (°C; mean ± SD)
36.4 ± 0.4
36.6 ± 0.8
36.7 ± 0.9
36.5 ± 0.5
WAZ (SD; median, range)
Underweight (WAZ < −2 SD, %)
MUAC (cm; mean ± SD)c
16.3 ± 1.3
15.9 ± 1.1
15.9 ± 1.2
15.9 ± 1.0
Clinically assessed malnutrition (%)
History within preceding 2 weeks (%)
Separating into mono-infections with P. falciparum and P. ovale, no significant differences with respect to the clinical manifestation were observed. One notable exception was anaemia, which tended to be more common among P. falciparum infected children (62.5%, 115/184) as compared to their P. ovale infected peers (45.7%, 19/35; P = 0.06).
In this cross-sectional survey among largely afebrile children attending school in southern Rwanda, 22% were infected with Plasmodium spp., predominantly P. falciparum, and 41% of these infections were submicroscopic. Infection was predicted by older age, low altitude, previous treatment, and absence of electricity or bicycle. Despite its ostensibly asymptomatic presentation, infection was associated with anaemia, underweight, and a number of non-specific symptoms in the children´s histories.
Infection prevalence varied greatly between locations, with a tendency of decline closer to the city of Butare (located in Ngoma sector, Fig. 1). Such variation is considered to reflect multiple and geographically diverse factors including altitude, rainfall, topography, land use, SES and health-seeking behaviour . Among these, altitude was the strongest predictor of infection in the present study. Declining infection prevalence with increasing altitude is a common pattern also in neighbouring countries [8, 28–31]. For example, in Tanzanian children, infection prevalence declined by 5–21% for every 100-m increase in altitude [28, 30]. The extent of this effect even on a small range between 1560 and 1850 m in the present study is remarkable. The higher prevalence of Plasmodium infection at lower altitude, i.e., usually at the valley bottoms, is most likely explained by the closer proximity to vector breeding sites in stagnant waters and commonly practiced rice cultivation. In line with that, valley floors in neighbouring Burundi are considered sources of infection at higher altitudes . Of note, the prevalence of Plasmodium infection did not show the usual decline with age, i.e., with cumulative exposure to malaria parasites. This suggests that the intensity of Plasmodium transmission in the study area is insufficient to build up immune responses during childhood, which prevent infection. Likewise, in neighbouring Tanzania, the age-dependent decline of infection prevalence has been found to vanish at an altitude similar to that of the present study area [28, 30]. In a previous survey in the study area , infection prevalence in young community children increased up to the age of 4–5 years and up to a level of 20–25%, i.e., a prevalence that was seen in most of the children in the present study. Children aged 10 years showed a significantly increased infection prevalence. This may reflect their comparatively lower use of bed nets as well as other behavioural characteristics of older children such as increased exposure due to longer staying outside in the evenings or traveling to areas of higher transmission. Similar to other studies from Rwanda [12, 18, 32], various indicators of low SES associated with Plasmodium infection but in multivariate analysis, only the protective parameters household availability of electricity and of a bicycle (each present in 16%) remained. While electricity reflects both, increased SES and proximity to rather urban settings , the latter may indicate increased access to malaria-related information, improved awareness and increased usage of curative services. Anti-malarial treatment, mostly with artemether-lumefantrine, within the preceding month was paradoxically associated with increased infection prevalence. This observation was also made among children under 5 years of age in the study area  and may stem from recurring parasitaemia following treatment. Drug resistance markers associated with reappearing parasitaemia following artemether-lumefantrine treatment are established in the study area [33, 34], but reports on actual treatment failure are absent. No information on the completeness of treatment was collected, i.e., on adherence to the 3-days regimen, but incomplete drug intake, e.g., in the community or after the first dose in a health facility, may have contributed to reappearing parasitaemia. Moreover, persisting gametocytes could have produced positive PCR results. Alternatively, this finding may reflect the aggregation of infections including rapid re-infections in a specifically vulnerable subset of the population .
Recent studies on malaria parasitaemia among school-age children have shown a prevalence range of <1 to 64% in East Africa and of 1 to 83% in West Africa . Symptomatic malaria contributes to school absenteeism, impaired cognition and reduced school performance [5, 10, 16, 20]. As for asymptomatic parasitaemia, predominating in the present study, findings are less clear-cut: in Yemen  and DR Congo , no influence on children´s cognition was observed while this was the case in Uganda , Mali , and Kenya . Overall, asymptomatic parasitaemia is abundant; it accounts for three in four infections detected in community settings , and many of those are submicroscopic [2, 3]. While some of these infections become symptomatic, most can remain asymptomatic for several months [37, 38]. Counterintuitively, the proportion of submicroscopic infections on a global scale is inversely correlated to transmission intensity, i.e., the proportion is highest at low endemicity [2, 3, 11, 21]. Asymptomatic carriers to a sizeable extent contribute to malaria transmission, also in low endemicity settings [2, 3, 7], even though the magnitude of this impact is not well established . In the present study, more than 90% of Plasmodium infected children attending school were considered asymptomatic based on the absence of fever. Nevertheless, anaemia prevalence was doubled in infected children and mean Hb reduced by 1 g/dL. This degree of Hb reduction is approximately half of that observed among younger and also largely asymptomatic children below 5 years in the study area . In addition, infected children more frequently had underweight and clinically assessed malnutrition than their non-infected peers. These findings argue against asymptomatic presentation but rather for a less obvious one. This is even more the case when considering the more frequently positive histories in infected as compared to non-infected children of fever, tiredness, weakness, poor appetite, abdominal pain, and vomiting. The influence of submicroscopic infections on the latter was similar to the one of microscopically visible parasitaemia but the effect on Hb levels was smaller and no association with underweight was observed. These findings add to the notion that “asymptomatic” Plasmodium infections do have subtle health consequences and should be regarded as potentially harmful . In fact, symptomless children infected with malaria parasites have been shown to have low-grade inflammation as well as evidence of platelet and endothelial activation, in addition to reduced Hb levels .
The present study has limitations. The definition of asymptomatic was based on a single-time point assessment; potentially, some children were in a pre-symptomatic period of parasitaemia at the moment of examination. Likewise, PCR detection of malaria parasites may include gametocytes, e.g., persisting after treatment. Submicroscopic parasitaemia may reflect both, very early and thus asymptomatic infections, which later on could develop to clinical malaria, and low level infections persisting in individuals able to control parasitaemia. Of note, studies in murine models have demonstrated that plasmodial DNA is rapidly removed from circulation after treatment or inoculation of killed parasites . Due to its design as a deworming study, questionnaire-based interviews focused on mainly general and gastrointestinal signs and symptoms whereas more intuitive malaria-related conditions, e.g., shivering or sweats, were not included. Although unfortunate, pronounced symptoms in this regard would likely have been detected upon clinical examination. Moreover, this circumstance enabled detecting significant associations of Plasmodium infection with less obvious “soft” symptoms. Plasmodium and Ascaris infections were not correlated and adjusting for the presence of the latter did not substantially affect the odds of symptoms due to the former. Infections with different Plasmodium species were combined since small numbers of non-falciparum malaria infections rendered separate analysis unsubstantial. In clinical terms, P. falciparum and P. ovale did not differ much, with the exception of a larger contribution to anaemia of the former.
The health and educational consequences of malaria and asymptomatic parasitaemia in school children as well as the role of this large group as a silent reservoir of infection are increasingly recognized [5, 9–20]. School children are not specifically targeted in malaria control programmes so far, and due to the predominance of seemingly asymptomatic parasitaemia, these infections usually remain untreated. The use of insecticide-treated nets (ITNs) in school children is notoriously low [5, 41] but schools are ideally suited to provide education on malaria, prevention and ITNs. This may include information on the importance of adhering to the 3-days regimen of artemisinin-based combination treatment. Schools also can serve as hubs for ITN distribution, and boarding schools in endemic areas should equip their dormitories with ITNs. Chemoprophylaxis or intermittent preventive treatment (IPT) have shown to reduce malaria-associated morbidity in school children [42, 43]. IPT, also termed intermittent parasite clearance in schools (IPCs) or seasonal malaria chemoprevention (SMC), needs to account for the epidemiological characteristics of malaria in a given area, and can greatly reduce the proportion of Plasmodium infected children (reviewed in Ref. ), and also improve sustained attention . School-based intermittent screening and treatment of malaria is another option but results have been disappointing so far . It remains to be elucidated, which drug-based regimen fits best and is most cost-effective for the different transmission settings across SSA. Integration with other school-based health promotion activities, e.g., routine deworming, is also desirable.
JBG, CB, KCS, and FPM designed the study. KCS, DG, CM, JCM, FH, KF, CB, JN, and FPM were responsible for patient recruitment, clinical and laboratory examinations. IU, CK, GSH, TA, and AS contributed to data interpretation. PM did the statistical analyses. KCS and FPM wrote the paper with major contributions of the other authors. All authors read and approved the final manuscript.
We thank the participating pupils, parents, teachers, community health workers, and UTHB laboratory staff. This work forms part of the doctoral theses of KCS and DG.
The authors declare that they have no competing interests.
Availability of data and material
Data supporting the findings will be available upon request sent to the corresponding author.
Ethics approval and consent to participate
Written informed consent and assent were obtained from the parents and the children, respectively. The study protocol was approved by the Rwandan National Ethics Committee (242/RNEC/2014), and the Rwandan Ministry of Education granted permission to conduct the study (2543/12.00/2014).
This study was supported by the German Federal Ministry of Education and Research (Grant 01DG13006A, MOPACUR). The sponsor had no role in study design; in the collection, analysis and interpretation of data; in the writing of the report; or in the decision to submit the article for publication.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
- White NJ, Pukrittayakamee S, Hien TT, Faiz MA, Mokuolu OA, Dondorp AM. Malaria. Lancet. 2014;383:723–35.View ArticlePubMedGoogle Scholar
- Bousema T, Okell L, Felger I, Drakeley C. Asymptomatic malaria infections: detectability, transmissibility and public health relevance. Nat Rev Microbiol. 2014;12:833–40.View ArticlePubMedGoogle Scholar
- Galatas B, Bassat Q, Mayor A. Malaria parasites in the asymptomatic: looking for the hay in the haystack. Trends Parasitol. 2016;32:296–308.View ArticlePubMedGoogle Scholar
- Greenwood BM. Asymptomatic malaria infections–do they matter? Parasitol Today. 1987;3:206–14.View ArticlePubMedGoogle Scholar
- Nankabirwa J, Brooker SJ, Clarke SE, Fernando D, Gitonga CW, Schellenberg D, et al. Malaria in school-age children in Africa: an increasingly important challenge. Trop Med Int Health. 2014;19:1294–309.View ArticlePubMedPubMed CentralGoogle Scholar
- Al Serouri AW, Grantham-McGregor SM, Greenwood B, Costello A. Impact of asymptomatic malaria parasitaemia on cognitive function and school achievement of schoolchildren in the Yemen Republic. Parasitology. 2000;121:337–45.View ArticlePubMedGoogle Scholar
- Alves FP, Gil LH, Marrelli MT, Ribolla PE, Camargo EP, Da Silva LH. Asymptomatic carriers of Plasmodium spp. as infection source for malaria vector mosquitoes in the Brazilian Amazon. J Med Entomol. 2005;42:777–9.View ArticlePubMedGoogle Scholar
- Baliraine FN, Afrane YA, Amenya DA, Bonizzoni M, Menge DM, Zhou G, et al. High prevalence of asymptomatic Plasmodium falciparum infections in a highland area of western Kenya: a cohort study. J Infect Dis. 2009;200:66–74.View ArticlePubMedPubMed CentralGoogle Scholar
- Clarke SE, Jukes MC, Njagi JK, Khasakhala L, Cundill B, Otido J, et al. Effect of intermittent preventive treatment of malaria on health and education in schoolchildren: a cluster-randomised, double-blind, placebo-controlled trial. Lancet. 2008;372:127–38.View ArticlePubMedPubMed CentralGoogle Scholar
- Nankabirwa J, Wandera B, Kiwanuka N, Staedke SG, Kamya MR, Brooker SJ. Asymptomatic Plasmodium infection and cognition among primary schoolchildren in a high malaria transmission setting in Uganda. Am J Trop Med Hyg. 2013;88:1102–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Okell LC, Bousema T, Griffin JT, Ouédraogo AL, Ghani AC, Drakeley CJ. Factors determining the occurrence of submicroscopic malaria infections and their relevance for control. Nat Commun. 2012;3:1237.View ArticlePubMedPubMed CentralGoogle Scholar
- Kateera F, Mens PF, Hakizimana E, Ingabire CM, Muragijemariya L, Karinda P, et al. Malaria parasite carriage and risk determinants in a rural population: a malariometric survey in Rwanda. Malar J. 2015;14:16.View ArticlePubMedPubMed CentralGoogle Scholar
- Laishram DD, Sutton PL, Nanda N, Sharma VL, Sobti RC, Carlton JM, et al. The complexities of malaria disease manifestations with a focus on asymptomatic malaria. Malar J. 2012;11:29.View ArticlePubMedPubMed CentralGoogle Scholar
- Walldorf JA, Cohee LM, Coalson JE, Bauleni A, Nkanaunena K, Kapito-Tembo A, et al. School-age children are a reservoir of malaria infection in Malawi. PLoS ONE. 2015;10:e0134061.View ArticlePubMedPubMed CentralGoogle Scholar
- de Mast Q, Brouwers J, Syafruddin D, Bousema T, Baidjoe AY, de Groot PG, et al. Is asymptomatic malaria really asymptomatic? Hematological, vascular and inflammatory effects of asymptomatic malaria parasitemia. J Infect. 2015;71:587–96.View ArticlePubMedGoogle Scholar
- Thuilliez J, Sissoko MS, Toure OB, Kamate P, Berthélemy JC, Doumbo OK. Malaria and primary education in Mali: a longitudinal study in the village of Donéguébougou. Soc Sci Med. 2010;71:324–34.View ArticlePubMedPubMed CentralGoogle Scholar
- Kurtzhals JA, Addae MM, Akanmori BD, Dunyo S, Koram KA, Appawu MA, et al. Anaemia caused by asymptomatic Plasmodium falciparum infection in semi-immune African schoolchildren. Trans R Soc Trop Med Hyg. 1999;93:623–7.View ArticlePubMedGoogle Scholar
- Gahutu JB, Steininger C, Shyirambere C, Zeile I, Cwinya-Ay N, Danquah I, et al. Prevalence and risk factors of malaria among children in southern highland Rwanda. Malar J. 2011;10:134.View ArticlePubMedPubMed CentralGoogle Scholar
- Ehrhardt S, Burchard GD, Mantel C, Cramer JP, Kaiser S, Kubo M, et al. Malaria, anemia, and malnutrition in African children—defining intervention priorities. J Infect Dis. 2006;194:108–14.View ArticlePubMedGoogle Scholar
- Fernando SD, Rodrigo C, Rajapakse S. The ‘hidden’ burden of malaria: cognitive impairment following infection. Malar J. 2010;9:366.View ArticlePubMedPubMed CentralGoogle Scholar
- Okell LC, Ghani AC, Lyons E, Drakeley CJ. Submicroscopic infection in Plasmodium falciparum-endemic populations: a systematic review and meta-analysis. J Infect Dis. 2009;200:1509–17.View ArticlePubMedGoogle Scholar
- Karema C, Aregawi MW, Rukundo A, Kabayiza A, Mulindahabi M, Fall IS, et al. Trends in malaria cases, hospital admissions and deaths following scale-up of anti-malarial interventions, 2000–2010, Rwanda. Malar J. 2012;11:236.View ArticlePubMedPubMed CentralGoogle Scholar
- Staudacher O, Heimer J, Steiner F, Kayonga Y, Havugimana JM, Ignatius R, et al. Soil-transmitted helminths in southern highland Rwanda: associated factors and effectiveness of school-based preventive chemotherapy. Trop Med Int Health. 2014;19:812–24.View ArticlePubMedGoogle Scholar
- World Health Organization. WHO AnthroPlus software. Geneva: World Health Organization. 2009. http://www.who.int/growthref/tools/en/. Accessed 31 Oct 2016.
- Sullivan KM, Mei Z, Grummer-Strawn L, Parvanta I. Haemoglobin adjustments to define anaemia. Trop Med Int Health. 2008;13:1267–71.View ArticlePubMedGoogle Scholar
- Rubio JM, Post RJ, van Leeuwen WM, Henry MC, Lindergard G, Hommel M. Alternative polymerase chain reaction method to identify Plasmodium species in human blood samples: the semi-nested multiplex malaria PCR (SnM-PCR). Trans R Soc Trop Med Hyg. 2002;96(Suppl 1):S199–204.View ArticlePubMedGoogle Scholar
- Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, et al. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg. 2011;84:338–43.View ArticlePubMedPubMed CentralGoogle Scholar
- Drakeley CJ, Carneiro I, Reyburn H, Malima R, Lusingu JP, Cox J, et al. Altitude-dependent and -independent variations in Plasmodium falciparum prevalence in northeastern Tanzania. J Infect Dis. 2005;191:1589–98.View ArticlePubMedGoogle Scholar
- Mboera LE, Kamugisha ML, Rumisha SF, Kisinza WN, Senkoro KP, Kitua AY. Malaria and mosquito net utilisation among schoolchildren in villages with or without healthcare facilities at different altitudes in Iringa District. Tanzania. Afr Health Sci. 2008;8:114–9.PubMedGoogle Scholar
- Bødker R, Msangeni HA, Kisinza W, Lindsay SW. Relationship between the intensity of exposure to malaria parasites and infection in the Usambara Mountains, Tanzania. Am J Trop Med Hyg. 2006;74:716–23.PubMedGoogle Scholar
- Protopopoff N, Van Bortel W, Marcotty T, Van Herp M, Maes P, Baza D, et al. Spatial targeted vector control in the highlands of Burundi and its impact on malaria transmission. Malar J. 2007;6:158.View ArticlePubMedPubMed CentralGoogle Scholar
- Bizimana JP, Twarabamenye E, Kienberger S. Assessing the social vulnerability to malaria in Rwanda. Malar J. 2015;14:2.View ArticlePubMedPubMed CentralGoogle Scholar
- Zeile I, Gahutu JB, Shyirambere C, Steininger C, Musemakweri A, Sebahungu F, et al. Molecular markers of Plasmodium falciparum drug resistance in southern highland Rwanda. Acta Trop. 2012;121:50–4.View ArticlePubMedGoogle Scholar
- Tacoli C, Gai PP, Bayingana C, Sifft K, Geus D, Ndoli J, et al. Artemisinin resistance-associated K13 polymorphisms of Plasmodium falciparum in southern Rwanda, 2010–2015. Am J Trop Med Hyg. 2016. doi:10.4269/ajtmh.16-0483.PubMedGoogle Scholar
- Woolhouse ME, Dye C, Etard JF, Smith T, Charlwood JD, Garnett GP, et al. Heterogeneities in the transmission of infectious agents: implications for the design of control programs. Proc Natl Acad Sci USA. 1997;94:338–42.View ArticlePubMedPubMed CentralGoogle Scholar
- Boivin MJ, Giordani B, Ndanga K, Maky MM, Manzeki KM, Ngunu N, et al. Effects of treatment for intestinal parasites and malaria on the cognitive abilities of schoolchildren in Zaire, Africa. Health Psychol. 1993;12:220–6.View ArticlePubMedGoogle Scholar
- Babiker HA, Abdel-Muhsin AM, Ranford-Cartwright LC, Satti G, Walliker D. Characteristics of Plasmodium falciparum parasites that survive the lengthy dry season in eastern Sudan where malaria transmission is markedly seasonal. Am J Trop Med Hyg. 1998;59:582–90.PubMedGoogle Scholar
- Roucher C, Rogier C, Dieye-Ba F, Sokhna C, Tall A, Trape JF. Changing malaria epidemiology and diagnostic criteria for Plasmodium falciparum clinical malaria. PLoS ONE. 2012;7:e46188.View ArticlePubMedPubMed CentralGoogle Scholar
- Lin JT, Ubalee R, Lon C, Balasubramanian S, Kuntawunginn W, Rahman R, et al. Microscopic Plasmodium falciparum gametocytemia and infectivity to mosquitoes in Cambodia. J Infect Dis. 2016;213:1491–4.View ArticlePubMedGoogle Scholar
- Jarra W, Snounou G. Only viable parasites are detected by PCR following clearance of rodent malarial infections by drug treatment or immune responses. Infect Immun. 1998;66:3783–7.PubMedPubMed CentralGoogle Scholar
- Noor AM, Kirui VC, Brooker SJ, Snow RW. The use of insecticide treated nets by age: implications for universal coverage in Africa. BMC Public Health. 2009;9:369.View ArticlePubMedPubMed CentralGoogle Scholar
- Geerligs PD, Brabin BJ, Eggelte TA. Analysis of the effects of malaria chemoprophylaxis in children on haematological responses, morbidity and mortality. Bull World Health Organ. 2003;81:205–16.PubMedPubMed CentralGoogle Scholar
- Meremikwu MM, Donegan S, Esu E. Chemoprophylaxis and intermittent treatment for preventing malaria in children. Cochrane Database Syst Rev. 2008;2:CD003756.Google Scholar
- Halliday KE, Okello G, Turner EL, Njagi K, Mcharo C, Kengo J, et al. Impact of intermittent screening and treatment for malaria among school children in Kenya: a cluster randomised trial. PLoS Med. 2014;11:e1001594.View ArticlePubMedPubMed CentralGoogle Scholar