The influence of the genetic complexity of Plasmodium falciparum infections on the epidemiology of malaria

Genetic characterization of malaria parasites in human blood stage infections has provided important insights into the genetics of Plasmodium falciparum populations and given rise to a field frequently referred to as 'molecular epidemiology'. This might be defined as the combination of parasite population genetic analysis with clinical and epidemiological analysis of a study population in order to achieve a better understanding of infection and immunity and long-term patterns of disease incidence and severity. Longitudinal studies on infection and clinical disease incidence, combined with improvements in the sensitivity of detection of low level, normally asymptomatic, parasite infections have formed an important part of this effort. Two molecular epidemiological studies of malaria under low and moderate intensity transmission, in Sudan and Ghana respectively, are reviewed here to illustrate how the parasite genotyping approaches based on deoxyribonucleic acid which Douglas Barker pioneered in the study of Leishmania have developed after their application to malaria research.     

Evaluating malaria attributable morbidity in endemic areas

There are no specific clinical signs or symptoms of malaria. Fever attacks, anemia, or signs of severity like coma or respiratory distress cannot easily be attributed to malaria in people who are infected most of the time. Ascribing clinical manifestations to malaria is problematic in populations that are regularly exposed to the transmission of human plasmodia. The more transmission is intense and regular, the higher the prevalence of asymptomatic infections. In areas of intense and perennial malaria transmission, more than 90% of the population may be infected and the simple detection of a plasmodial infection is not enough to attribute clinical manifestations to malaria. Naturally acquired anti-malaria immunity permitting asymptomatic infections is incomplete and temporary. It is an obstacle to the estimation of the malaria burden in endemic areas. The positive association between parasite density and fever allows the attribution of clinical attacks to malaria. The relationship between parasitaemia and the risk of fever is not continuous. An age- and endemicity-dependent threshold effect of parasite density has been demonstrated and can be used to distinguish clinical attacks due to malaria from others. Clinical diagnosis and evaluation of malaria are problematic in three situations: in public health to estimate the malaria burden for health services, in clinical research to evaluate treatments or prophylactic measures (drug, vaccine, anti-vectorial devices), and in basic research on pathophysiology, immunology or genetic susceptibility to clinical malaria. No one diagnostic definition nor procedure for detecting cases is adequate for all three purposes. Case detection may be passive (in health structures for example) or active (in population). The choice of methods for diagnosis and recruitment depends on the objectives and whether a "pragmatic" or "explicative" approach is used. The radical differences between these approaches are often unsuspected or ignored.     

Complement activation in experimental human malaria infection

The objective of this study was to investigate complement activation in uncomplicated, early phases of human malaria. Fifteen healthy volunteers were experimentally infected with Plasmodium falciparum malaria. Parasitemia and complement activation products were assessed. During blood stage parasitemia, volunteers showed a significant increase in soluble terminal complement complex (TCC) formation. After start of a curative regimen of artemether/lumefantrine, TCC further increased due to activation of both the classical and the alternative pathway. In-vitro studies confirmed activation of complement by parasite cultures. We thus detected an increase in complement activation in volunteers with experimentally induced malaria, even before parasitemia could be detected microscopically. This significant increase in complement activation occurred despite the possible control of TCC formation by complement regulatory proteins on erythrocytes and the extremely low levels of parasitemia. Treatment with artemether/lumefantrine was followed by classical and alternative pathway complement activation, without evidence for mannan-binding-lectin-mediated complement activation.     

Iron and malaria: absorption, efficacy and safety

Febrile malaria and asymptomatic malaria parasitemia substantially decrease iron absorption in single-meal, stable isotope studies in women and children, but to date there is no evidence of decreased efficacy of iron-fortified foods in malaria-endemic regions. Without inadequate malarial surveillance or health care, giving iron supplements to children in areas of high transmission could increase morbidity and mortality. The most likely explanation is the appearance of non-transferrin-bound iron (NTBI) in the plasma. NTBI forms when the rate of iron influx into the plasma exceeds the rate of iron binding to transferrin. Two studies in women have reported substantially increased NTBI with the ingestion of iron supplements. Our studies confirm this, but found no significant increase in NTBI on consumption of iron-fortified food. It seems likely that the malarial parasite in hepatocytes can utilize NTBI, but it cannot do so in infected erythrocytes. NTBI however may increase the sequestration of parasite-infected erythrocytes in capillaries. Bacteremia is common in children with severe malaria and sequestration in villi capillaries could lead to a breaching of the intestinal barrier, allowing the passage of pathogenic bacteria into the systemic circulation. This is especially important as frequent high iron doses increase the number of pathogens in the intestine at the expense of the barrier bacteria.     

Changing patterns of autochthonous malaria transmission in the United States: a review of recent outbreaks.

Three recent outbreaks of locally acquired malaria in densely populated areas of the United States demonstrate the continued risk for mosquitoborne transmission of this disease. Increased global travel, immigration, and the presence of competent anopheline vectors throughout the continental United States contribute to the ongoing threat of malaria transmission. The likelihood of mosquitoborne transmission in the United States is dependent on the interactions between the human host, anopheline vector, malaria parasite, and environmental conditions. Recent changes in the epidemiology of locally acquired malaria and possible factors contributing to these changes are discussed.     

Human malaria: Segregation analysis of blood infection levels in a suburban area and a rural area in Burkina Faso

The genetic control of blood infection levels in human malaria remains unclear. Case control studies have not demonstrated a strong association between candidate genes and blood parasite densities as opposed to surveys that have focused on severe malaria. As an alternative approach, we used segregation analyses to determine the genetic control of blood parasitemia. We surveyed 509 residents (53 pedigrees) in a rural area and 389 residents (41 pedigrees) in an urban area during 18 months. Each family was visited 20 times and 28 times in the urban area and in the rural area; the mean number of parasitemia measurements per subject was 12.1 in the town and 14.9 in the village. The intensity of transmission of Plasmodium falciparum was 8-fold higher in the rural area than in the urban area. Using the class D regressive model for both populations, we found that blood parasite densities were correlated between sibs. We obtained strong evidence for a major effect, but we found that the transmission of this major effect was not compatible with a simple Mendelian model, suggesting a more complex mode of inheritance. Moreover, there was a strong interaction between major effect and age, suggesting that the influence of the putative major gene may be more prominent in children than in adults. Further nonparametric linkage studies, such as sib pair analysis, that focus on children would help us better understand the genetic control of blood infection levels. Genet. Epidemiol. 15:435–450,1998. © 1998 Wiley-Liss, Inc.     

Virulence in rodent malaria: host genotype by parasite genotype interactions.

In an effort to understand what limits the virulence of malaria parasites, we infected inbred mice of three genotypes (C57Bl/6J, CBA/Ca and DBA/2) with one of two parasite lines of the rodent malaria Plasmodium chabaudi. One of these parasite lines had been serially passaged through C57Bl/6J mice and had evolved higher asexual growth rate, virulence and transmission in the process. The other parasite line was the unadapted ancestral line which had low virulence. In all three host genotypes, the C57Bl/6J-adapted parasite line was more virulent than the ancestral line thus indicating that trade-offs in virulence between alternative host genotypes had not placed strong constraints on the evolution of high virulence in this system. By examining the infection dynamics for fitness-related components-asexual parasite population growth, transmission and virulence-we revealed alternative possible explanations for what sets the upper limit to virulence in nature. The total number of transmission forms (gametocytes) produced during the infection, a measure of parasite Darwinian fitness, was four-fold higher in mice that survived the infection than those which died. Among mice that survived, total gametocyte production was greatest in the host genotype that suffered intermediate levels of morbidity (anaemia and weight loss). Thus, there were transmission costs of high virulence that were partly due to host death (as most theoretical models of virulence evolution assume), but perhaps partly due to some factor related to high morbidity. Both mortality and morbidity-related factors might therefore influence the upper limit on virulence of malaria parasites.     

Clinical epidemiology of malaria in the highlands of western Kenya

Malaria in the highlands of Kenya is traditionally regarded as unstable and limited by low temperature. Brief warm periods may facilitate malaria transmission and are therefore able to generate epidemic conditions in immunologically naive human populations living at high altitudes. The adult:child ratio (ACR) of malaria admissions is a simple tool we have used to assess the degree of functional immunity in the catchment population of a health facility. Examples of ACR are collected from inpatient admission data at facilities with a range of malaria endemicities in Kenya. Two decades of inpatient malaria admission data from three health facilities in a high-altitude area of western Kenya do not support the canonical view of unstable transmission. The malaria of the region is best described as seasonal and meso-endemic. We discuss the implications for malaria control options in the Kenyan highlands.     

Bayesian analysis of an epidemiologic model of Plasmodium falciparum malaria infection in Ndiop, Senegal

Plasmodium falciparum has a complex transmission cycle. Public health planning and research would benefit from the ability of a calibrated model to predict the epidemiologic characteristics of populations living in areas of malaria endemicity. This paper describes the application of Bayesian calibration to a malaria transmission model using longitudinal data gathered from 176 subjects in Ndiop, Senegal, from July 1, 1993, to July 31, 1994. The model was able to adequately predict P. falciparum parasitemia prevalence in the study population. Further insight into the dynamics of malaria in Ndiop was provided. During the dry season, the estimated fraction of nonimmune subjects goes down to 20% and then increases up to 80%. The model-predicted time-weighted average incidences contributed by nonimmune and immune individuals are 0.52 cases per day and 0.47 cases per day, respectively. The median times needed to acquire infection (conversion delay) for nonimmune and immune individuals are estimated at 39 days and 285 days, respectively.     

A return of endemic malaria to the Netherlands is highly unlikely]

The Netherlands has been free from malaria since the early 1960, due to a combination of factors: active search and treatment of patients and parasite carriers, targeted use of insecticides, changes in farming and in housing of man and cattle, pollution of surface water with phosphates and the fact that surface waters became fresher. These factors reduced the mosquito population that is dependent on brackish water. The Dutch malaria mosquito cannot transmit the parasite of tropical malaria. The mosquito population could possibly increase due to measures to 'develop nature' but the number of parasite carriers, the acute disease manifestations, the quality and organization of the health care system make it extremely unlikely that local transmission will occur. Fears that malaria may become endemic and that the population in the western parts of the country will have to apply malaria chemoprophylaxis in the near future, are unfounded.     

Unstable malaria in Sudan: the influence of the dry season. Clone multiplicity of Plasmodium falciparum infections in individuals exposed to variable levels of disease transmission.

Studies of infection and immunity to malaria often take little account of the fact that the amount of infectious challenge individuals receive is very variable. Classic studies in areas of holoendemic transmission showed that clinical immunity develops quite rapidly during childhood, although the processes through which increasing levels of resistance to infection are acquired are still not understood. However, holoendemic transmission is one end of the spectrum of malaria epidemiology and the development of clinical immunity is also affected by factors such as the infection rate and the local parasite species composition. An exceptionally simple type of malaria transmission occurs during the short, autumnal malaria outbreaks of the Sudanese sahel-savannah belt, where a sparse 200-500 mm of rain falls in 2-3 summer months, Plasmodium falciparum causes > 95% of malaria cases in most areas, and the entomological inoculation rate (EIR) is very low by African standards; thus the population dynamics of malaria parasites are less affected by super-infection. A comparison of certain features of parasite genetic diversity, particularly the average number of parasite clones present in infections in the Sudanese sahel and in malaria study sites with different levels of transmission, is presented. It is proposed that increasing EIRs are associated with progressively smaller increases in the average number of malaria parasite clones per host and the implications of this relationship for studies on malaria infection and immunity are discussed.     

Permethrin-impregnated curtains in malaria control

The impact of permethrin-impregnated curtains on the incidence of malaria episodes, parasitaemia and splenomegaly was assessed during a 22 month period in 2 groups of children aged 0.5-6 years. One group lived in houses where permethrin-impregnated curtains had been installed, the other group lived in houses without curtains. A significant reduction of incidence of malaria episodes, mean parasite density, parasite prevalence and splenomegaly was consistently observed in the intervention group towards the end of the period of moderate transmission, whereas no clear-cut impact could be demonstrated during the high transmission period. The influence of malaria pressure and community utilization on the protective efficiency of curtains is discussed. Because of their acceptability and the ease of reimpregnation, curtains proved to be a suitable technique for integration into primary health care.     

Clearing asymptomatic parasitaemia increases the specificity of the definition of mild febrile malaria

In clinical trials, the specificity of the disease endpoint is critical to an accurate estimate of vaccine efficacy. We used a logistic regression model to analyse parasite densities among children before and after treatment with antimalarials, in order to estimate the impact that clearing asymptomatic parasitaemia had on the specificity of the endpoint of febrile malaria. The malaria attributable fever fraction was higher after antimalarial treatment (i.e. fever and parasitaemia were more likely to be causally related), implying that drug treatment prior to monitoring decreased the misclassification of febrile malaria. In intervention studies with febrile malaria as an endpoint, clearing asymptomatic parasitaemia increases the study's power more effectively than raising the threshold parasitaemia.     

A cohort study of Plasmodium falciparum diversity during the dry season in Ndiop, a Senegalese village with seasonal, mesoendemic malaria

Prolonged carriage of Plasmodium falciparum in humans during the dry season is critical for parasite survival, as the infected subjects constitute a major reservoir in the absence of transmission. Yet, very little is known about the host/parasite interactions contributing to parasite persistence. In order to study the characteristics of P. falciparum infections during the dry season, we have genotyped parasites collected from untreated, asymptomatic individuals during 3 cross-sectional surveys conducted during the dry season in Ndiop, a Senegalese village with seasonal, mesoendemic malaria. Monthly entomological surveillance did not detect any transmission during that period. Parasite prevalence decreased markedly in the children aged < 7 years after 7 months of undetected transmission, but was stable in older children and adults throughout the dry season. In all chronically infected individuals, infection complexity remained stable, but there were substantial fluctuations of individual genotype(s), reflecting complex dynamics of multiple-clone infections during chronic asymptomatic parasite carriage. This fluctuation resulted in changes in the msp1 and msp2 allelic distribution within the cohort after 7 months of undetected transmission, contrasting with the stability observed during the preceding rainy season in that village.     

Potential impact of global climate change on malaria risk.

The biological activity and geographic distribution of the malarial parasite and its vector are sensitive to climatic influences, especially temperature and precipitation. We have incorporated General Circulation Model-based scenarios of anthropogenic global climate change in an integrated linked-system model for predicting changes in malaria epidemic potential in the next century. The concept of the disability-adjusted life years is included to arrive at a single measure of the effect of anthropogenic climate change on the health impact of malaria. Assessment of the potential impact of global climate change on the incidence of malaria suggests a widespread increase of risk due to expansion of the areas suitable for malaria transmission. This predicted increase is most pronounced at the borders of endemic malaria areas and at higher altitudes within malarial areas. The incidence of infection is sensitive to climate changes in areas of Southeast Asia, South America, and parts of Africa where the disease is less endemic; in these regions the numbers of years of healthy life lost may increase significantly. However, the simulated changes in malaria risk must be interpreted on the basis of local environmental conditions, the effects of socioeconomic developments, and malaria control programs or capabilities.     

State transition detection in the spatio-temporal incidence of malaria

Mosquito-borne disease spread might exhibit irregular epidemic fronts caused by ecological heterogeneity in the risk factors. To determine Plasmodium vivax infection spread in north-eastern Venezuela, we used the State Transition Index (STI) to detect the spatial locations of malaria incidence boundaries and their dynamics over time. Then, we evaluated the role of population size on disease persistence. Boundary locations of malaria were found to be highly spatially variable. Waves of infection were observed in the spatial mosaics of large and small nearby localities due to a strong asynchrony in the epidemic dynamics between both host populations. Our results suggest that the epidemic spatial diffusion follows a hierarchy from large, populated villages (with few or no seasonal parasite fadeouts) to smaller, less populated localities, where infection outbreak was irregular or disease dynamics showed frequent fadeouts. Our findings stress the importance of malaria surveillance and control in these larger communities.     

Importance and prevention of malaria in pregnancy

Malaria in pregnancy is one of the most important preventable causes of low birthweight deliveries worldwide. It is also a major cause of severe maternal anaemia contributing to maternal mortality. It is estimated that 40% of the world's pregnant women are exposed to malaria infection during pregnancy. The clinical features of Plasmodium falciparum malaria in pregnancy depend to a large extent on the immune status of the woman, which in turn is determined by her previous exposure to malaria. In pregnant women with little or no pre-existing immunity, such as women from non-endemic areas or travellers to malarious areas, infection is associated with high risks of severe disease with maternal and perinatal mortality. Women are at particular risk of cerebral malaria, hypoglycaemia, pulmonary oedema and severe haemolytic anaemia. Fetal and perinatal loss has been documented to be as high as 60-70% in non-immune women with malaria. Adults who are long-term residents of areas of moderate or high malaria transmission, including large parts of sub-Saharan Africa, usually have a high level of immunity to malaria. Infection is frequently asymptomatic and severe disease is uncommon. During pregnancy this immunity to malaria is altered. Infection is still frequently asymptomatic, so may go unsuspected and undetected, but is associated with placental parasitization. Malaria in pregnancy is a common cause of severe maternal anaemia and low birthweight babies, these complications being more common in primigravidae than multigravidae. Preventative strategies include regular chemoprophylaxis, intermittent preventative treatment with antimalarials and insecticide-treated bednets.     

Age reduces resistance and tolerance in malaria-infected mice

Once infected, hosts can rely on two strategies to cope with parasites: fight them (resist the infection) or minimize the damage they induce (tolerate the infection). While there is evidence that aging reduces resistance, how tolerance varies as hosts become old has been barely studied. Here, we used a rodent malaria parasite (Plasmodium yoelii) to investigate whether 2- and 12-month old house mice differ in their capacity to resist and tolerate the infection. We found that 12-month old mice harbored higher parasitemia, showing that age reduces resistance to malaria. Infection-induced deterioration of host health was assessed using red blood cell and body mass loss. Using both traits, the rate of decline in host health, as parasitemia increased, was more pronounced in 12- than in 2-month old mice, showing that age is also associated with impaired tolerance to malaria. Overall, resistance and tolerance positively covaried; however, this was only due to the age effect, since, within age classes, the two traits were not correlated. These results show that senescing individuals might be both more susceptible to infectious diseases and less able to cope with the damage that infection induces.     

疟疾疫苗研究进展

疟疾是一种世界范围内的传染病,严重影响人类的身体健康和生命安全.疫苗作为控制乃至消灭传染病的有效手段,在疟疾研究中受到广泛关注.目前针对疟原虫生活史各期的期特异性疫苗、传播阻断型疫苗、多阶段/多抗原疫苗以及减毒活疫苗都处于研究中.尽管尚无成熟疫苗推入市场,但一些候选疫苗已进入临床实验,并产生了非常有希望的结果.该文就疟疾疫苗的研究进展做一综述.     

Congenital malaria. Parasitological and serological studies in Niamey (Niger)

Congenital malaria is defined as the presence of Plasmodium parasites in the erythrocytes of newborns less than seven days old. The aim of this study was to determine the incidence of congenital malaria and its possible clinical consequences. We carried out a prospective survey in Niamey, the capital of Niger (600,000 inhabitants) from July to September 1993. Niamey is in an area of mesoendemic malaria and this period of the year corresponds to the rainy season, when malaria transmission is maximal. Ninety mothers and their newborns were included. We assessed the clinical status of the mother and child at the time of the delivery, and took blood smears to check for parasitemia and blood samples to check for antimalaria antibodies by indirect immunofluorescence (IIF). The placenta was not examined. Clinical signs of malaria (fever, splenomegaly, anemia and jaundice) were absent in all mothers and children and 88 of the 90 children had normal birth weights. Plasmodium falciparum was the only parasite detected, with 49 of the 90 mothers and 12 of the 90 newborns having positive blood smears. Serological tests detected the presence of antimalaria antibodies in 73 of the 90 mothers (81.1%) and 68 of the 90 newborns (75.5%). Thus, we found no cases of congenital malaria with clinical signs in this study, despite the high frequency of parasites and antimalaria antibodies. The reasons for this absence of cases of congenital malaria with symptoms are discussed.