Sarah E. Reece’s research while affiliated with University of Edinburgh and other places

The discovery of rhythmicity in host and pathogen activities dates back to the Hippocratic era, but the causes and consequences of these biological rhythms have remained poorly understood. Rhythms in infection phenotypes or traits are observed across taxonomically diverse hosts and pathogens, suggesting general evolutionary principles. Understanding these principles may enable rhythms to be leveraged in manners that improve drug and vaccine efficacy or disrupt pathogen timekeeping to reduce virulence and transmission. Explaining and exploiting rhythms in infections require an integrative and multidisciplinary approach, which is a hallmark of research within chronobiology. Many researchers are welcomed into chronobiology from other fields after observing an unexpected rhythm or time-of-day effect in their data. Such findings can launch a rich new research topic, but engaging with the concepts, approaches and dogma in a new discipline can be daunting. Fortunately, chronobiology has well-developed frameworks for interrogating rhythms that can be readily applied in novel contexts. Here, we provide a ‘how to’ guide for exploring unexpected daily rhythms in infectious disease research. We outline how to establish: whether the rhythm is circadian, to what extent the host and pathogen are responsible, the relevance for host–pathogen interactions, and how to explore therapeutic potential. This article is part of the Theo Murphy meeting issue ‘Circadian rhythms in infection and immunity’.

Chronobiology is a multidisciplinary field that extends across the tree of life, transcends all scales of biological organization, and has huge translational potential. For the UK to harness the opportunities presented within applied chronobiology, we need to build our network outwards to reach stakeholders that can directly benefit from our discoveries. In this article, we discuss the importance of biological rhythms to our health, society, economy and environment, with a particular focus on circadian rhythms. We subsequently introduce the vision and objectives of BioClocks UK, a newly formed research network, whose mission is to stimulate researcher interactions and sustain discovery-impact cycles between chronobiologists, wider research communities and multiple industry sectors. This article is part of the Theo Murphy meeting issue ‘Circadian rhythms in infection and immunity’.

The environments that parasites experience within hosts change dramatically over 24 h. How rhythms shape host–parasite–vector interactions is poorly understood owing to the challenges of disentangling the roles of rhythms of multiple interacting species in the context of the complex lifecycles of parasites. Using canonical circadian clock-disrupted hosts, we probe the limits of flexibility in the rhythmic replication of malaria (Plasmodium) parasites and quantify the consequences for fitness proxies of both parasite and host. We reveal that parasites alter the duration of their replication rhythm to resonate with host rhythms that have short (21 h) daily T-cycles as accurately as when infecting hosts with 24 h cycles, but appear less capable of extending their replication rhythm in hosts with long (27 h) cycles. Despite matching the period of short T-cycle hosts, parasites are unable to lock to the correct phase, likely leading to lower within-host productivity and a reduction in transmission potential. However, parasites in long T-cycle hosts do not experience substantial fitness costs. Furthermore, T-cycle duration does not affect disease severity in clock-disrupted hosts. Understanding the rhythmic replication of malaria parasites offers the opportunity to interfere with parasite timing to improve health and reduce transmission. This article is part of the Theo Murphy meeting issue issue ‘Circadian rhythms in infection and immunity’.

Parasites rely on exploiting resources from their hosts and vectors for survival and transmission. This includes nutritional resources, which vary in availability between different hosts and changes during infections. For malaria (Plasmodium) parasites, sexual reproduction (sporogony) and subsequent development of oocysts, which produce sporozoites infectious to the vertebrate host, occurs in the mosquito vector. Mosquitoes in the field exhibit diversity in the amount and type of food they acquire, directly impacting the nutrients available for the replication and development of parasites. While the rate of parasite transmission from vector to host is influenced by the nutritional state of mosquitoes, whether this is due to resource limitation mediating parasite development and productivity is poorly understood. We use the rodent model parasite P. chabaudi and the vector Anopheles stephensi to ask how variation in the amount of sugar and blood provided to malaria-infected mosquitoes affects the potential for parasites to transmit from vector to host. We show that parasites in well-resourced mosquitoes reach a larger oocyst size earlier in development, suggesting faster growth, and have a 1.7-fold higher sporozoite burden than parasites whose vectors only receive sugar. However, this increase in productivity is only partly explained by oocyst development, suggesting that resource availability also impacts the ability of sporozoites to reach the salivary glands. This challenges the assumption of a simple relationship between the number or size of oocysts and onward transmission potential. Furthermore, our findings suggest malaria parasites may actively adjust oocyst growth rate to best exploit nutritional resources; while parasites in low-resourced mosquitoes exhibited a reduction in oocyst burden during sporogony, the remaining oocysts developed more rapidly in the later stages of oocyst development, catching up to reach a similar size to those in well-resourced mosquitoes. Understanding the impacts of resource availability for malaria transmission is urgent given that parasites encounter increasingly variable vectors as consequences of climate change and vector control tools.

Undertaking certain activities at the time of day that maximises fitness is assumed to explain the evolution of circadian clocks. Organisms often use daily environmental cues such as light and food availability to set the timing of their clocks. These cues may be the environmental rhythms that ultimately determine fitness, act as proxies for the timing of less tractable ultimate drivers, or are used simply to maintain internal synchrony. While many pathogens/parasites undertake rhythmic activities, both the proximate and ultimate drivers of their rhythms are poorly understood. Explaining the roles of rhythms in infections offers avenues for novel interventions to interfere with parasite fitness and reduce the severity and spread of disease. Here, we perturb several rhythms in the hosts of malaria parasites to investigate why parasites align their rhythmic replication to the host's feeding‐fasting rhythm. We manipulated host rhythms governed by light, food or both, and assessed the fitness implications for parasites, and the consequences for hosts, to test which host rhythms represent ultimate drivers of the parasite's rhythm. We found that alignment with the host's light‐driven rhythms did not affect parasite fitness metrics. In contrast, aligning with the timing of feeding‐fasting rhythms may be beneficial for the parasite, but only when the host possess a functional canonical circadian clock. Because parasites in clock‐disrupted hosts align with the host's feeding‐fasting rhythms and yet derive no apparent benefit, our results suggest cue(s) from host food act as a proxy rather than being a key selective driver of the parasite's rhythm. Alternatively, parasite rhythmicity may only be beneficial because it promotes synchrony between parasite cells and/or allows parasites to align to the biting rhythms of vectors. Our results also suggest that interventions can disrupt parasite rhythms by targeting the proxies or the selective factors driving them without impacting host health.

Malaria infections represent an iconic example of developmental synchrony, where periodic fevers can result when the population of parasites develops synchronously within host red blood cells. The level of synchrony appears to vary across individual hosts and across parasite species and strains, variation that—once quantified—can illuminate the ecological and evolutionary drivers of synchrony. Yet current approaches for quantifying synchrony in parasites are either biased by population dynamics or unsuitable when population growth rates vary through time, features ubiquitous to parasite populations in vitro and in vivo . Here we develop an approach to estimate synchrony that accounts for population dynamics, including changing population growth rates, and validate it with simulated time series data encompassing a range of synchrony levels in two different host-parasite systems: malaria infections of mice and human malaria parasites in vitro . This new method accurately quantifies developmental synchrony from per capita growth rates using obtainable abundance data even with realistic sampling noise, without the need to sort parasites into developmental stages. Our approach enables variability in developmental schedules to be disentangled from even extreme variation in population dynamics, providing a comparative metric of developmental synchrony.

Most malaria (Plasmodium spp.) parasite species undergo asexual replication synchronously within the red blood cells of their vertebrate host. Rhythmicity in this intraerythrocytic developmental cycle (IDC) enables parasites to maximise exploitation of the host and align transmission activities with the time of day that mosquito vectors blood feed. The IDC is also responsible for the major pathologies associated with malaria, and plasticity in the parasite's rhythm can confer tolerance to antimalarial drugs. Both the severity of infection (virulence) and synchrony of the IDC vary across species and between genotypes of Plasmodium; however, this variation is poorly understood. The theory predicts that virulence and IDC synchrony are negatively correlated, and we tested this hypothesis using two closely related genotypes of the rodent malaria model Plasmodium chabaudi that differ markedly in virulence. We also test the predictions that, in response to perturbations to the timing (phase) of the IDC schedule relative to the phase of host rhythms (misalignment), the virulent parasite genotype recovers the correct phase relationship faster, incurs less fitness losses and so hosts benefit less from misalignment when infected with a virulent genotype. Our predictions are partially supported by results suggesting that the virulent parasite genotype is less synchronous in some circumstances and recovers faster from misalignment. While hosts were less anaemic when infected by misaligned parasites, the extent of this benefit did not depend on parasite virulence. Overall, our results suggest that interventions to perturb the alignment between the IDC schedule, and host rhythms and increase synchrony between parasites within each IDC, could alleviate disease symptoms. However, virulent parasites, which are better at withstanding conventional antimalarial treatment, would also be intrinsically better able to tolerate such interventions.

Malaria infections represent an iconic example of developmental synchrony, where periodic fevers can result when the population of parasites develops synchronously within host red blood cells. The level of synchrony appears to vary across individual hosts and across parasite species and strains, variation that--once quantified--can illuminate the ecological and evolutionary drivers of synchrony. Yet current approaches for quantifying synchrony in parasites are either biased by population dynamics or unsuitable when population growth rates vary through time, features ubiquitous to parasite populations in vitro and in vivo. Here we develop an approach to estimate synchrony that accounts for population dynamics, including changing population growth rates, and validate it with simulated time series data encompassing a range of synchrony levels in two different host-parasite systems: malaria infections of mice and human malaria parasites in vitro. This new method accurately quantifies developmental synchrony from per capita growth rates using obtainable abundance data even with realistic sampling noise, without the need to sort parasites into developmental stages. Our approach enables variability in developmental schedules to be disentangled from even extreme variation in population dynamics, providing a comparative metric of developmental synchrony.

Circadian rhythms are ubiquitous in nature and endogenous circadian clocks drive the daily expression of many fitness-related behaviors. However, little is known about whether such traits are targets of selection imposed by natural enemies. In Hawaiian populations of the nocturnally active Pacific field cricket (Teleogryllus oceanicus), males sing to attract mates, yet sexually selected singing rhythms are also subject to natural selection from the acoustically orienting and deadly parasitoid fly, Ormia ochracea. Here, we use T. oceanicus to test whether singing rhythms are endogenous and scheduled by circadian clocks, making them possible targets of selection imposed by flies. We also develop a novel audio-to-circadian analysis pipeline, capable of extracting useful parameters from which to train machine learning algorithms and process large quantities of audio data. Singing rhythms fulfilled all criteria for endogenous circadian clock control, including being driven by photoschedule, self-sustained periodicity of approximately 24 h, and being robust to variation in temperature. Furthermore, singing rhythms varied across individuals, which might suggest genetic variation on which natural and sexual selection pressures can act. Sexual signals and ornaments are well-known targets of selection by natural enemies, but our findings indicate that the circadian timing of those traits’ expression may also determine fitness.

Background The extrinsic incubation period (EIP), defined as the time it takes for malaria parasites in a mosquito to become infectious to a vertebrate host, is one of the most influential parameters for malaria transmission but remains poorly understood. The EIP is usually estimated by quantifying salivary gland sporozoites in subsets of mosquitoes, which requires terminal sampling. However, assays that allow repeated sampling of individual mosquitoes over time could provide better resolution of the EIP. Methods We tested a non-destructive assay to quantify sporozoites of two rodent malaria species, Plasmodium chabaudi and Plasmodium berghei, expelled throughout 24-h windows, from sugar-soaked feeding substrates using quantitative-PCR. Results The assay is able to quantify sporozoites from sugar-soaked feeding substrates, but the prevalence of parasite-positive substrates was low. Various methods were attempted to increase the detection of expelled parasites (e.g. running additional technical replicates; using groups rather than individual mosquitoes), but these did not increase the detection rate, suggesting that expulsion of sporozoites is variable and infrequent. Conclusions We reveal successful detection of expelled sporozoites from sugar-soaked feeding substrates. However, investigations of the biological causes underlying the low detection rate of sporozoites (e.g. mosquito feeding behaviour, frequency of sporozoite expulsion or sporozoite clumping) are needed to maximise the utility of using non-destructive assays to quantify sporozoite dynamics. Increasing detection rates will facilitate the detailed investigation on infection dynamics within mosquitoes, which is necessary to explain the highly variable EIP of Plasmodium and to improve understanding of malaria transmission dynamics. Graphical Abstract