Chronic malaria infections increase family inequalities and reduce parental fitness: experimental evidence from a wild bird population

Avian malaria parasites (Plasmodium) occur commonly in wild birds and are an increasingly popular model system for understanding host–parasite co‐evolution. However, whether these parasites have fitness consequences for hosts in endemic areas is much debated, particularly since wild‐caught individuals almost always harbour chronic infections of very low parasite density. We used the anti‐malarial drug Malarone^TM to test experimentally for fitness effects of chronic malaria infection in a wild population of breeding blue tits (Cyanistes caeruleus). Medication caused a pronounced reduction in Plasmodium infection intensity, usually resulting in complete clearance of these parasites from the blood, as revealed by quantitative PCR. Positive effects of medication on malaria‐infected birds were found at multiple stages during breeding, with medicated females showing higher hatching success, provisioning rates and fledging success compared to controls. Most strikingly, we found that treatment of maternal malaria infections strongly altered within‐family differences, with reduced inequality in hatching probability and fledging mass within broods reared by medicated females. These within‐brood effects appear to explain higher fledging success among medicated females and are consistent with a model of parental optimism in which smaller (marginal) offspring can be successfully raised to independence if additional resources become available during the breeding attempt. Overall, these results demonstrate that chronic avian malaria infections, far from being benign, can have significant effects on host fitness and may thus constitute an important selection pressure in wild bird populations.     

Effect of repeated exposure to Plasmodium relictum (lineage SGS1) on infection dynamics in domestic canaries

Parasites are known to exert strong selection pressures on their hosts and, as such, favour the evolution of defence mechanisms. The negative impact of parasites on their host can have substantial consequences in terms of population persistence and the epidemiology of the infection. In natural populations, however, it is difficult to assess the cost of infection while controlling for other potentially confounding factors. For instance, individuals are repeatedly exposed to a variety of parasite strains, some of which can elicit immunological memory, further protecting the host from subsequent infections. Cost of infection is, therefore, expected to be particularly strong for primary infections and to decrease for individuals surviving the first infectious episode that are re-exposed to the pathogen. We tested this hypothesis experimentally using avian malaria parasites (Plasmodium relictum-lineage SGS1) and domestic canaries (Serinus canaria) as a model. Hosts were infected with a controlled dose of P. relictum as a primary infection and control birds were injected with non-infected blood. The changes in haematocrit and body mass were monitored during a 20 day period. A protein of the acute phase response (haptoglobin) was assessed as a marker of the inflammatory response mounted in response to the infection. Parasite intensity was also monitored. Surviving birds were then re-infected 37 days post primary infection. In agreement with the predictions, we found that primary infected birds paid a substantially higher cost in terms of infection-induced reduction in haematocrit compared with re-exposed birds. After the secondary infection, re-exposed hosts were also able to clear the infection at a faster rate than after the primary infection. These results have potential consequences for the epidemiology of avian malaria, since birds re-exposed to the pathogen can maintain parasitemia with low fitness costs, allowing the persistence of the pathogen within the host population.     

Dynamics of parasitemia of malaria parasites in a naturally and experimentally infected migratory songbird, the great reed warbler Acrocephalus arundinaceus

Little is known about the development of infection of malaria parasites of the genus Plasmodium in wild birds. We used qPCR, targeting specific mitochondrial lineages of Plasmodium ashfordi (GRW2) and Plasmodium relictum (GRW4), to monitor changes in intensities of parasitemia in captive great reed warblers Acrocephalus arundinaceus from summer to spring. The study involved both naturally infected adults and experimentally infected juveniles. The experiment demonstrated that P. ashfordi and P. relictum lineages differ substantially in several life-history traits (e.g. prepatent period and dynamics of parasitemia) and that individual hosts show substantial differences in responses to these infections. The intensity of parasitemia of lineages in mixed infections co-varied positively, suggesting a control mechanism by the host that is general across the parasite lineages. The intensity of parasitemia for individual hosts was highly repeatable suggesting variation between the host individuals in their genetic or acquired control of the infections. In future studies, care must be taken to avoid mixed infections in wild caught donors, and when possible use mosquitoes for the experiments as inoculation of infectious blood ignores important initial stages of the contact between the bird and the parasite.     

Malaria infection negatively affects feather growth rate in the house sparrow Passer domesticus

Birds often face various stressors during feather renewing, for example, enduring infection with blood parasites. Because nutritional resources are typically limited, especially for wild animals, when an individual allocates energy to one physiological system, there is subsequently less for other processes, thereby requiring a trade‐off. Surprisingly, potential trade‐offs between malaria infection and feather growth rate have not been experimentally considered yet. Here, we conducted three studies to investigate whether a trade‐off occurs among feather growth rate, malaria infection and host health conditions. First, we explored whether naturally infected and uninfected house sparrows differed in feather growth rate in the wild. Second, we asked whether experimental inoculation of malaria parasites and/or forcing the renewal of a tail feather. Lastly, we evaluated whether individual condition was affected by experimentally‐induced feather regrowth and/or malaria experimental infection. Our findings showed that feather growth rate was negatively affected by natural malaria infection status in free‐living birds and by experimental infection in captive birds. Furthermore, birds that did not increase body mass or hematocrit during the experimental study had slower feather growth. Together our results suggest that infection with blood parasites has more negative health effects than the growth of tail feathers and that these two processes (response to blood parasite infection and renewal of feathers) are traded‐off against each other. As such, our results highlight the role of malaria parasites as a potential mechanism driving other trade‐offs in wild passerines.     

Evolution of Plastic Transmission Strategies in Avian Malaria

Malaria parasites have been shown to adjust their life history traits to changing environmental conditions. Parasite relapses and recrudescences—marked increases in blood parasite numbers following a period when the parasite was either absent or present at very low levels in the blood, respectively—are expected to be part of such adaptive plastic strategies. Here, we first present a theoretical model that analyses the evolution of transmission strategies in fluctuating seasonal environments and we show that relapses may be adaptive if they are concomitant with the presence of mosquitoes in the vicinity of the host. We then experimentally test the hypothesis that Plasmodium parasites can respond to the presence of vectors. For this purpose, we repeatedly exposed birds infected by the avian malaria parasite Plasmodium relictum to the bites of uninfected females of its natural vector, the mosquito Culex pipiens, at three different stages of the infection: acute (∼34 days post infection), early chronic (∼122 dpi) and late chronic (∼291 dpi). We show that: (i) mosquito-exposed birds have significantly higher blood parasitaemia than control unexposed birds during the chronic stages of the infection and that (ii) this translates into significantly higher infection prevalence in the mosquito. Our results demonstrate the ability of Plasmodium relictum to maximize their transmission by adopting plastic life history strategies in response to the availability of insect vectors.     

Signs of a vector's adaptive choice: on the evasion of infectious hosts and parasite‐induced mortality

Laboratory and field experiments have demonstrated in many cases that malaria vectors do not feed randomly, but show important preferences either for infected or non‐infected hosts. These preferences are likely in part shaped by the costs imposed by the parasites on both their vertebrate and dipteran hosts. However, the effect of changes in vector behaviour on actual parasite transmission remains a debated issue. We used the natural associations between a malaria‐like parasite Polychromophilus murinus, the bat fly Nycteribia kolenatii and a vertebrate host the Daubenton's bat Myotis daubentonii to test the vector's feeding preference based on the host's infection status using two different approaches: 1) controlled behavioural assays in the laboratory where bat flies could choose between a pair of hosts; 2) natural bat fly abundance data from wild‐caught bats, serving as an approximation of realised feeding preference of the bat flies. Hosts with the fewest infectious stages of the parasite were most attractive to the bat flies that did switch in the behavioural assay. In line with the hypothesis of costs imposed by parasites on their vectors, bat flies carrying parasites had higher mortality. However, in wild populations, bat flies were found feeding more based on the bat's body condition, rather than its infection level. Though the absolute frequency of host switches performed by the bat flies during the assays was low, in the context of potential parasite transmission they were extremely high. The decreased survival of infected bat flies suggests that the preference for less infected hosts is an adaptive trait. Nonetheless, other ecological processes ultimately determine the vector's biting rate and thus transmission. Inherent vector preferences therefore play only a marginal role in parasite transmission in the field. The ecological processes rather than preferences per se need to be identified for successful epidemiological predictions.     

Testing Local Adaptation in a Natural Great Tit-Malaria System: An Experimental Approach

Finding out whether Plasmodium spp. are coevolving with their vertebrate hosts is of both theoretical and applied interest and can influence our understanding of the effects and dynamics of malaria infection. In this study, we tested for local adaptation as a signature of coevolution between malaria blood parasites, Plasmodium spp. and its host, the great tit, Parus major. We conducted a reciprocal transplant experiment of birds in the field, where we exposed birds from two populations to Plasmodium parasites. This experimental set-up also provided a unique opportunity to study the natural history of malaria infection in the wild and to assess the effects of primary malaria infection on juvenile birds. We present three main findings: i) there was no support for local adaptation; ii) there was a male-biased infection rate; iii) infection occurred towards the end of the summer and differed between sites. There were also site-specific effects of malaria infection on the hosts. Taken together, we present one of the few experimental studies of parasite-host local adaptation in a natural malaria system, and our results shed light on the effects of avian malaria infection in the wild.     

Diversification and host switching in avian malaria parasites

The switching of parasitic organisms to novel hosts, in which they may cause the emergence of new diseases, is of great concern to human health and the management of wild and domesticated populations of animals. We used a phylogenetic approach to develop a better statistical assessment of host switching in a large sample of vector-borne malaria parasites of birds (Plasmodium and Haemoproteus) over their history of parasite-host relations. Even with sparse sampling, the number of parasite lineages was almost equal to the number of avian hosts. We found that strongly supported sister lineages of parasites, averaging 1.2% sequence divergence, exhibited highly significant host and geographical fidelity. Event-based matching of host and parasite phylogenetic trees revealed significant cospeciation. However, the accumulated effects of host switching and long distance dispersal cause these signals to disappear before 4% sequence divergence is achieved. Mitochondrial DNA nucleotide substitution appears to occur about three times faster in hosts than in parasites, contrary to findings on other parasite-host systems. Using this mutual calibration, the phylogenies of the parasites and their hosts appear to be similar in age, suggesting that avian malaria parasites diversified along with their modern avian hosts. Although host switching has been a prominent feature over the evolutionary history of avian malaria parasites, it is infrequent and unpredictable on time scales germane to public health and wildlife management.     

Avian malaria is associated with increased reproductive investment in the blue tit

Haemosporidians causing avian malaria are very common parasites among bird species. Their negative effects have been repeatedly reported in terms of deterioration in survival prospects or reproductive success. However, a positive association between blood parasites and avian fitness has also been reported. Here, we studied a relationship between presence of malaria parasites and reproductive performance of the host, a hole‐breeding passerine – the blue tit Cyanistes caeruleus. Since the malaria parasites might affect their hosts differently depending on environmental conditions, we performed brood size manipulation experiment to differentiate parental reproductive effort and study the potential interaction between infection status and brood rearing conditions on reproductive performance. We found individuals infected with malaria parasites to breed later in the season in comparison with uninfected birds, but no differences were detected in clutch size. Interestingly, infected parents produced heavier and larger offspring with stronger reaction to phytohemagglutinin. More importantly, we found a significant interaction between infection status and brood size manipulation in offspring tarsus length and reaction to phytohemagglutinin: presence of parasites had stronger positive effect among birds caring for experimentally enlarged broods. Our results might be interpreted either in the light of the parasite‐mediated selection or terminal investment hypothesis.     

Does avian malaria reduce fledging success: an experimental test of the selection hypothesis

Like many parasites, avian haematozoa are often found at lower infection intensities in older birds than young birds. One explanation, known as the “selection” hypothesis, is that infected young birds die before reaching adulthood, thus removing the highest infection intensities from the host population. We tested this hypothesis in the field by experimentally infecting nestling rock pigeons (Columba livia) with the malaria parasite Haemoproteus columbae. We compared the condition and fledging success of infected nestlings to that of uninfected controls. There was no significant difference in the body mass, fledging success, age at fledging, or post-fledging survival of experimental versus control birds. These results were unexpected, given that long-term studies of older pigeons have demonstrated chronic effects of H. columbae. We conclude that H. columbae has little impact on nestling pigeons, even when they are directly infected with the parasite. Our study provides no support for the selection hypothesis that older birds have lower parasite loads because parasites are removed from the population by infected nestlings dying. To our knowledge, this is the first study to test the impact of avian malaria using experimental inoculations under natural conditions.     

Molecular characterization of malarial parasites in captive passerine birds

Seven of 28 passerine birds that died in captivity were positive for malarial parasites by polymerase chain reaction targeting the mitochondrial cytochrome b (cytB) and apicoplast ribosomal RNA (rRNA) genes. Each bird was infected with a single parasite lineage having a unique genotype. Apicoplast rRNA sequences were present both in Haemoproteus spp. and Plasmodium spp. and had typically high adenosine + thymidine content. Phylogenies for cytB and apicoplast rRNA sequences were largely congruent and supported previous studies that suggest that Plasmodium-Haemoproteus spp. underwent synchronous speciation with their avian hosts, interrupted by sporadic episodes of host switching. Apicoplast phylogeny further indicated that Haemoproteus spp. are ancestral to Plasmodium spp. All the 7 infected passerine birds had histologic lesions of malaria, and malarial parasites may have contributed to the death of at least 4 animals. These findings provide new genetic data on passerine hematozoa, including initial sequences of apicoplast DNA, and emphasize the relevance of parasite prevalence, evolutionary relationships, and host switching to modern management and husbandry practices of captive birds.     

Malaria parasite pre-erythrocytic stage infection: gliding and hiding

In malaria, the red blood cell-infectious form of the Plasmodium parasite causes illness and the possible death of infected hosts. The initial infection in the liver caused by the mosquito-borne sporozoite parasite stage, however, causes little pathology and no symptoms. Nevertheless, pre-erythrocytic parasite stages are attracting passionate research efforts not least because they are the most promising targets for malaria vaccine development. Here, we review how the infectious sporozoite makes its way to the liver and subsequently develops within hepatocytes. We discuss the factors, both parasite and host, involved in the interactions that occur during this "silent" phase of infection.     

Environmental stress increases the prevalence and intensity of blood parasite infection in the common lizard Lacerta vivipara

Parasites affect the life-histories and fitness of their hosts. It has been demonstrated that the ability of the immune system to cope with parasites partly depends on environmental conditions. In particular, stressful conditions have an immunosuppressive effect and may affect disease resistance. The relationship between environmental stress and parasitism was investigated using a blood parasite of the common lizard Lacerta vivipara. In laboratory cages, density and additional stressors had a significant effect on the intensity of both natural parasitaemia and parasitaemia induced by experimental infection. Four weeks after infection, crowded lizards had three times more parasites than noncrowded lizards. After 1 month of stress treatment, naturally infected lizards had a significantly higher level of plasma corticosterone and a higher parasite load than nonstressed individuals. In seminatural enclosures, stress induced by the habitat quality affected both the natural blood parasite prevalence and the intensity of parasitaemia of the host.     

Effects of malaria double infection in birds: one plus one is not two

Avian malaria parasites are supposed to exert negative effects on host fitness because these intracellular parasites affect host metabolism. Recent advances in molecular genotyping and microscopy have revealed that coinfections with multiple parasites are frequent in bird-malaria parasite systems. However, studies of the fitness consequences of such double infections are scarce and inconclusive. We tested if the infection with two malaria parasite lineages has more negative effects than single infection using 6 years of data from a natural population of house martins. Survival was negatively affected by both types of infections. We found an additive cost from single to double infection in body condition, but not in reproductive parameters (double-infected had higher reproductive success). These results demonstrate that malaria infections decrease survival, but also have different consequences on the breeding performance of single- and double-infected wild birds.     

Epizootiology of Plasmodium hermani in Florida: chronicity of experimental infections in domestic turkeys and northern bobwhites

A pen-reared northern bobwhite and a domestic turkey were infected with a strain of Plasmodium hermani obtained originally from a wild turkey in southern Florida. Blood films from these 2 birds were positive microscopically for 188 and 370 days postinfection (PI), respectively. Culicine mosquitoes (Culex nigripalpus and C. salinarius) were blood fed on the bobwhite and the turkey at different times during the infection and used to transmit the malaria to other bobwhites and turkeys up to days 298 and 473 PI, respectively. It was concluded that in nature, P. hermani could remain in a chronic phase in avian hosts for a year, or longer, allowing survival of the parasite between seasons of mosquito transmission.     

Effect of haemosporidian infections on host survival and recapture rate in the blue tit

Parasites are ubiquitous in the wild and by imposing fitness costs on their hosts they constitute an important selection factor. One of the most common parasites of wild birds are Plasmodium and Haemoproteus, protozoans inhabiting the blood, which cause avian malaria and malaria‐like disease, respectively. Although they are expected to cause negative effects in infected individuals, in many cases studies in natural populations failed to detect such effect. Using data from seven breeding seasons (2008–2014), we applied a multistate capture–mark–recapture approach to study the effect of infection with malaria and malaria‐like parasites, individual age and sex on the probability of survival and recapture rate in a small passerine, the blue tit Cyanistes caeruleus, inhabiting the island of Gotland, Sweden. We found no effect of infection on survival prospects. However, the recapture rate of infected individuals was higher than that of uninfected ones. Thus, while our data do not support the presence of infection costs in terms of host survival, it suggests that parasites from the genera Plasmodium and Haemoproteus may affect some aspects of host behaviour, which translates into biased estimation of infection frequency at the population level.     

Haemosporidian parasitism in the blackcap Sylvia atricapilla in relation to spring arrival and body condition

Parasites can exert strong selection on hosts. Throughout the year migrants are exposed to different sets of parasites, which may affect life history traits such as migratory schedules. Here, we studied the relationship between parasite infection and arrival date of blackcaps Sylvia atricapilla to their breeding grounds in Germany throughout a period of six years (2007–2012). We used two data sets, one that included all blackcaps and one that included only recaptured birds. We assesed whether parasites influence spring arrival to breeding grounds, and for the recaptured data set, we analysed temporal variation in parasitism (i.e. infection status and parasitaemia) throughout the breeding season. We used both microscopy and PCR (a fragment of ? 479 bp of the mtDNA cyt b) to determine haemosporidian infection. Blackcaps were mostly infected with Haemoproteus parabelopolskyi (lineages SYAT01 and SYAT02). Infection status, but not parasitaemia, was constant through time for individual birds; meaning that once a bird is infected, it most likely will retain the infection for life. We found that infection by haemosporidian parasites has no relationship to arrival date in this blackcap population; however, infection by H. parabelopolskyi has a marginally significant effect on arrival date of recaptured blackcaps, somewhat delaying their arrival to breeding grounds. Birds captured later in the season were more likely to be infected than those from early spring, and parasitaemia was frequently lower in birds captured earlier in the season compared to those captured later (summer).     

Malaria infection increases bird attractiveness to uninfected mosquitoes

The epidemiology of vector‐borne pathogens is largely determined by the host‐choice behaviour of their vectors. Here, we investigate whether a Plasmodium infection renders the host more attractive to host‐seeking mosquitoes. For this purpose, we work on a novel experimental system: the avian malaria parasite Plasmodium relictum, and its natural vector, the mosquito Culex pipiens. We provide uninfected mosquitoes with a choice between an uninfected bird and a bird undergoing either an acute or a chronic Plasmodium infection. Mosquito choice is assessed by microsatellite typing of the ingested blood. We show that chronically infected birds attract significantly more vectors than either uninfected or acutely infected birds. Our results suggest that malaria parasites manipulate the behaviour of uninfected vectors to increase their transmission. We discuss the underlying mechanisms driving this behavioural manipulation, as well as the broader implications of these effects for the epidemiology of malaria.     

Cross-species regulation of malaria parasitaemia in the human host

Longitudinal genetic analysis of the composition of malaria parasites infecting humans has demonstrated that individuals living in endemic areas are chronically infected with multiple genotypes and species of Plasmodium. The accumulation of infections is a consequence of superinfection from the bites of many infected anopheline mosquitoes. The clinical outcome of infection is determined by the host's ability to regulate the density of malaria parasites in the blood. Interestingly, most infections do not cause symptoms of malarial disease after a degree of immunity is acquired. Here, we review data from the first genetic study of the longitudinal dynamics of multiple Plasmodium species and genotypes in humans. The data show that the total parasite density of Plasmodium species oscillates around a threshold and that peaks of infection with each species do not coincide. We propose that malaria parasitaemia is controlled in a density-dependent manner in these semi-immune children. This implies that a cross-species mechanism of parasite regulation exists. A model of how multiple immune responses could act in concert to explain these within host dynamics is discussed in relation to known regulatory mechanisms.