Temporal dynamics of freshwater planktonic parasites inferred using a DNA metabarcoding time-series.

Parasites are important components of biodiversity and contributors to ecosystem functioning, but are often neglected in ecological studies. Most studies examine model parasite systems or single taxa, thus our understanding of community composition is lacking. Here, the seasonal and annual dynamics of parasites was quantified using a 5-year metabarcoding time-series of freshwater plankton, collected weekly. We first identified parasites in the dataset using literature searches of the taxonomic match and using sequence metadata from the National Center for Biotechnology Information (NCBI) nucleotide database. In total, 441 amplicon sequence variants (belonging to 18 phyla/clades) were classified as parasites. The four phyla/clades with the highest relative read abundance and richness were Chytridiomycota, Dinoflagellata, Oomycota and Perkinsozoa. Relative read abundance of total parasite taxa, Dinoflagellata and Perkinsozoa significantly varied with season and was highest in summer. Parasite richness varied significantly with season and year, and was generally lowest in spring. Each season had distinct parasite communities, and the difference between summer and winter communities was most pronounced. Combining DNA metabarcoding with searches of the literature and NCBI metadata allowed us to characterize parasite diversity and community dynamics and revealed the extent to which parasites contribute to the diversity of freshwater plankton communities.

Trophic position, elemental ratios and nitrogen transfer in a planktonic host-parasite-consumer food chain including a fungal parasite.

Parasitism is arguably the most commonly occurring consumer strategy. However, only a few food web studies assess how well stable isotopes reflect the trophic position of parasitic consumers and results are variable. Even fewer studies have measured the nutrient transfer by parasitic consumers, hindering an assessment of their role in nutrient transfer through food webs. Here we used a food chain consisting of a diatom as host, a chytrid as its parasitic consumer and a rotifer as the predatory consumer of the chytrid, to assess the trophic position of all three food-chain components using their natural 13C and 15N isotope signatures, and to measure the nitrogen transfer from the host via the chytrid to the rotifer by tracing 15N of a labelled host up the food chain. Additionally, we measured the carbon to nitrogen (C:N) ratios of all food-chain components. Natural isotope abundance results showed no clear 15N enrichment in the chytrid or rotifer relative to the primary producer. However, estimates of nitrogen transfer indicated that about 14% of host nitrogen was transferred per day from host to chytrid during infection epidemics and that some of this nitrogen was also transferred onward to the rotifer. Moreover, C:N ratios decreased with trophic level, suggesting that the chytrid provided a high-quality food source to the rotifer. In conclusion, our results support the "mycoloop". The mycooloop proposes that chytrid infections allow the transfer of nutrients bound in large, inedible phytoplankton to zooplankton through the production of edible transmission spores, thereby rerouting nutrients back into the food web.

Integrating chytrid fungal parasites into plankton ecology: research gaps and needs.

Chytridiomycota, often referred to as chytrids, can be virulent parasites with the potential to inflict mass mortalities on hosts, causing e.g. changes in phytoplankton size distributions and succession, and the delay or suppression of bloom events. Molecular environmental surveys have revealed an unexpectedly large diversity of chytrids across a wide range of aquatic ecosystems worldwide. As a result, scientific interest towards fungal parasites of phytoplankton has been gaining momentum in the past few years. Yet, we still know little about the ecology of chytrids, their life cycles, phylogeny, host specificity and range. Information on the contribution of chytrids to trophic interactions, as well as co-evolutionary feedbacks of fungal parasitism on host populations is also limited. This paper synthesizes ideas stressing the multifaceted biological relevance of phytoplankton chytridiomycosis, resulting from discussions among an international team of chytrid researchers. It presents our view on the most pressing research needs for promoting the integration of chytrid fungi into aquatic ecology.

Winter severity determines functional trait composition of phytoplankton in seasonally ice-covered lakes.

How climate change will affect the community dynamics and functionality of lake ecosystems during winter is still little understood. This is also true for phytoplankton in seasonally ice-covered temperate lakes which are particularly vulnerable to the presence or absence of ice. We examined changes in pelagic phytoplankton winter community structure in a north temperate lake (Müggelsee, Germany), covering 18 winters between 1995 and 2013. We tested how phytoplankton taxa composition varied along a winter-severity gradient and to what extent winter severity shaped the functional trait composition of overwintering phytoplankton communities using multivariate statistical analyses and a functional trait-based approach. We hypothesized that overwintering phytoplankton communities are dominated by taxa with trait combinations corresponding to the prevailing winter water column conditions, using ice thickness measurements as a winter-severity indicator. Winter severity had little effect on univariate diversity indicators (taxon richness and evenness), but a strong relationship was found between the phytoplankton community structure and winter severity when taxon trait identity was taken into account. Species responses to winter severity were mediated by the key functional traits: motility, nutritional mode, and the ability to form resting stages. Accordingly, one or the other of two functional groups dominated the phytoplankton biomass during mild winters (i.e., thin or no ice cover; phototrophic taxa) or severe winters (i.e., thick ice cover; exclusively motile taxa). Based on predicted milder winters for temperate regions and a reduction in ice-cover durations, phytoplankton communities during winter can be expected to comprise taxa that have a relative advantage when the water column is well mixed (i.e., need not be motile) and light is less limiting (i.e., need not be mixotrophic). A potential implication of this result is that winter severity promotes different communities at the vernal equinox, which may have different nutritional quality for the next trophic level and ecosystem-scale effects.

Hot summers raise public awareness of toxic cyanobacterial blooms.

Water quality of eutrophic lakes is threatened by harmful cyanobacterial blooms, which are favored by summer heatwaves and expected to intensify with global warming. Societal demands on surface water for drinking, irrigation and recreation are also highest in summer, especially during dry and warm conditions. Here, we analyzed trends in online searches to investigate how public awareness of cyanobacterial blooms is impacted by temperature in nine different countries over almost twenty years. Our findings reveal large seasonal and interannual variation, with more online searches for harmful cyanobacteria in temperate regions during hot summers. Online searches and media attention increased even more steeply with temperature than the incidence of cyanobacterial blooms, presumably because lakes attract more people during warm weather. Overall, our study indicates that warmer summers not only increase cyanobacterial bloom incidence, but also lead to a pronounced increase of the public awareness of toxic cyanobacterial blooms.

Herbicides in the environment alter infection dynamics in a microbial host-parasite system.

Parasites play an important role in the regulation of host population growth. How these ubiquitous stressors interact with anthropogenic stressors is less often studied. In a full factorial experiment we explored the independent and combined effects of the widely used herbicide diuron and a chytrid parasite on the fitness of genetically different monoclonal diatom populations. Furthermore, we evaluated how herbicide exposure influenced infection dynamics, parasite fitness and the impact of infectious disease on host populations. We found no evidence of host genetic variation for diuron sensitivity and parasite resistance. Instead, host population phenotype was a decisive factor in controlling parasite growth. Although herbicide exposure initially posed a constraint on disease transmission, it enhanced the spread of disease over time. Consequently, the nature of the parasite-toxicant stressor interaction shifted from antagonistic (on exponential host growth) towards additive (on final uninfected host density). We conclude that herbicide exposure can modify infection dynamics and impact of disease on host populations through the complex interplay between host and parasite growth dynamics and host population phenotype.

Moving beyond standard toxicological metrics: The effect of diclofenac on planktonic host-parasite interactions.

Pharmaceuticals are increasingly released into surface waters and therefore ubiquitous in aquatic systems. While pharmaceuticals are known to influence species interactions, their effect on host-parasite interactions is still underexplored despite potential ecosystem-level consequences. Here, we ask whether diclofenac, a widely used non-steroid anti-inflammatory drug, affects the interaction between a phytoplankton host (Staurastrum sp.; green alga) and its obligate fungal parasite (Staurastromyces oculus; chytrid fungus). We hypothesized that the effect of increasing diclofenac concentration on the host-parasite system depends on parasite exposure. We assessed acute and chronic effects of a wide range of diclofenac concentrations (0-150 mg/L) on host and parasite performance using a replicated long gradient design in batch cultures. Overall system response summarizing parameters related to all biotic components in an experimental unit i.e., number of bacteria and phytoplankton host cells along with photosynthetic yield (a measure of algal cell fitness), depended on diclofenac concentration and presence/absence of parasite. While host standing biomass decreased at diclofenac concentrations >10 mg/L in non-parasite-exposed treatments, it increased at ≥10 mg/L in parasite-exposed treatments since losses due to infection declined. During acute phase (0-48 h), diclofenac concentrations <0.1 mg/L had no effect on host net-production neither in parasite-exposed nor non-parasite-exposed treatments, but parasite infection ceased at 10 mg/L. During chronic phase (0-216 h), host net-production declined only at concentrations >10 mg/L in non-parasite-exposed cultures, while it was overall close to zero in parasite-exposed cultures. Our results suggest that chytrid parasites are more sensitive to diclofenac than their host, allowing a window of opportunity for growth of phytoplankton hosts, despite exposure to a parasite. Our work provides a first understanding about effects of a pharmaceutical on a host-parasite interaction beyond those defined by standard toxicological metrics.

Environmental refuges from disease in host-parasite interactions under global change.

The physiological performance of organisms depends on their environmental context, resulting in performance-response curves along environmental gradients. Parasite performance-response curves are generally expected to be broader than those of their hosts due to shorter generation times and hence faster adaptation. However, certain environmental conditions may limit parasite performance more than that of the host, thereby providing an environmental refuge from disease. Thermal disease refuges have been extensively studied in response to climate warming, but other environmental factors may also provide environmental disease refuges which, in turn, respond to global change. Here, we (1) showcase laboratory and natural examples of refuges from parasites along various environmental gradients, and (2) provide hypotheses on how global environmental change may affect these refuges. We strive to synthesize knowledge on potential environmental disease refuges along different environmental gradients including salinity and nutrients, in both natural and food-production systems. Although scaling up from single host-parasite relationships along one environmental gradient to their interaction outcome in the full complexity of natural environments remains difficult, integrating host and parasite performance-response can serve to formulate testable hypotheses about the variability in parasitism outcomes and the occurrence of environmental disease refuges under current and future environmental conditions.

Domestic dog demographic structure and dynamics relevant to rabies control planning in urban areas in Africa: the case of Iringa, Tanzania.

BACKGROUND: Mass vaccinations of domestic dogs have been shown to effectively control canine rabies and hence human exposure to rabies. Knowledge of dog population demography is essential for planning effective rabies vaccination programmes; however, such information is still rare for African domestic dog populations, particularly so in urban areas. This study describes the demographic structure and population dynamics of a domestic dog population in an urban sub-Saharan African setting. In July to November 2005, we conducted a full household-level census and a cross-sectional dog demography survey in four urban wards of Iringa Municipality, Tanzania. The achievable vaccination coverage was assessed by a two-stage vaccination campaign, and the proportion of feral dogs was estimated by a mark-recapture transect study. RESULTS: The estimated size of the domestic dog population in Iringa was six times larger than official town records assumed, however, the proportion of feral dogs was estimated to account for less than 1% of the whole population. An average of 13% of all households owned dogs which equalled a dog:human ratio of 1:14, or 0.31 dogs per household or 334 dogs km-2. Dog female:male ratio was 1:1.4. The average age of the population was 2.2 years, 52% of all individuals were less than one year old. But mortality within the first year was high (72%). Females became fertile at the age of 10 months and reportedly remained fertile up to the age of 11 years. The average number of litters whelped per fertile female per year was 0.6 with an average of 5.5 pups born per litter. The population growth was estimated at 10% y-1. CONCLUSIONS: Such high birth and death rates result in a rapid replacement of anti-rabies immunised individuals with susceptible ones. This loss in herd immunity needs to be taken into account in the design of rabies control programmes. The very small proportion of truly feral dogs in the population implies that vaccination campaigns aimed at the owned dog population are sufficient to control rabies in urban Iringa, and the same may be valid in other, comparable urban settings.

Phytoplankton responses to repeated pulse perturbations imposed on a trend of increasing eutrophication.

While eutrophication remains one of the main pressures acting on freshwater ecosystems, the prevalence of anthropogenic and nature-induced stochastic pulse perturbations is predicted to increase due to climate change. Despite all our knowledge on the effects of eutrophication and stochastic events operating in isolation, we know little about how eutrophication may affect the response and recovery of aquatic ecosystems to pulse perturbations. There are multiple ways in which eutrophication and pulse perturbations may interact to induce potentially synergic changes in the system, for instance, by increasing the amount of nutrients released after a pulse perturbation. Here, we performed a controlled press and pulse perturbation experiment using mesocosms filled with natural lake water to address how eutrophication modulates the phytoplankton response to sequential mortality pulse perturbations; and what is the combined effect of press and pulse perturbations on the resistance and resilience of the phytoplankton community. Our experiment showed that eutrophication increased the absolute scale of the chlorophyll-a response to pulse perturbations but did not change the proportion of the response relative to its pre-event condition (resistance). Moreover, the capacity of the community to recover from pulse perturbations was significantly affected by the cumulative effect of sequential pulse perturbations but not by eutrophication itself. By the end of the experiment, some mesocosms could not recover from pulse perturbations, irrespective of the trophic state induced by the press perturbation. While not resisting or recovering any less from pulse perturbations, phytoplankton communities from eutrophying systems showed chlorophyll-a levels much higher than non-eutrophying ones. This implies that the higher absolute response to stochastic pulse perturbations in a eutrophying system may increase the already significant risks for water quality (e.g., algal blooms in drinking water supplies), even if the relative scale of the response to pulse perturbations between eutrophying and non-eutrophying systems remains the same.

Making waves: Lessons learned from the COVID-19 anthropause in the Netherlands on urban aquatic ecosystem services provisioning and management.

The anomalous past two years of the COVID-19 pandemic have been a test of human response to global crisis management as typical human activities were significantly altered. The COVID-instigated anthropause has illustrated the influence that humans and the biosphere have on each other, especially given the variety of national mobility interventions that have been implemented globally. These local COVID-19-era restrictions influenced human-ecosystem interactions through changes in accessibility of water systems and changes in ecosystem service demand. Four urban aquatic case studies in the Netherlands demonstrated shifts in human demand during the anthropause. For instance, reduced boat traffic in Amsterdam canals led to improved water clarity. In comparison, ongoing service exploitation from increased recreational fishing, use of bathing waters and national parks visitation are heightening concerns about potential ecosystem degradation. We distilled management lessons from both the case studies as well as from recent literature pertaining to ecological intactness and social relevance. Equally important to the lessons themselves, however, is the pace at which informed management practices are established after the pandemic ends, particularly as many communities currently recognize the importance of aquatic ecosystems and are amenable to their protection.

The potential of zooplankton in constraining chytrid epidemics in phytoplankton hosts.

Fungal diseases threaten natural and man-made ecosystems. Chytridiomycota (chytrids) infect a wide host range, including phytoplankton species that form the basis of aquatic food webs and produce roughly half of Earth's oxygen. However, blooms of large or toxic phytoplankton form trophic bottlenecks, as they are inedible to zooplankton. Chytrids infecting inedible phytoplankton provide a trophic link to zooplankton by producing edible zoospores of high nutritional quality. By grazing chytrid zoospores, zooplankton may induce a trophic cascade, as a decreased zoospore density will reduce new infections. Conversely, fewer infections will not produce enough zoospores to sustain long-term zooplankton growth and reproduction. This intricate balance between zoospore density necessary for zooplankton energetic demands (growth/survival), and the loss in new infections (and thus new zoospores) because of grazing was tested empirically. To this end, we exposed a cyanobacterial host (Planktothrix rubescens) infected by a chytrid (Rizophydium megarrhizum) to a grazer density gradient (the rotifer Keratella cf. cochlearis). Rotifers survived and reproduced on a zoospore diet, but the Keratella population growth was limited by the amount of zoospores provided by chytrid infections, resulting in a situation where zooplankton survived but were restricted in their ability to control disease in the cyanobacterial host. We subsequently developed and parameterized a dynamical food-chain model using an allometric relationship for clearance rate to assess theoretically the potential of different-sized zooplankton groups to restrict disease in phytoplankton hosts. Our model suggests that smaller-sized zooplankton may have a high potential to reduce chytrid infections on inedible phytoplankton. Together, our results point out the complexity of three-way interactions between hosts, parasites, and grazers and highlight that trophic cascades are not always sustainable and may depend on the grazer's energetic demand.

Changes in N:P Supply Ratios Affect the Ecological Stoichiometry of a Toxic Cyanobacterium and Its Fungal Parasite.

Human activities have dramatically altered nutrient fluxes from the landscape into receiving waters. As a result, not only the concentration of nutrients in surface waters has increased, but also their elemental ratios have changed. Such shifts in resource supply ratios will alter autotroph stoichiometry, which may in turn have consequences for higher trophic levels, including parasites. Here, we hypothesize that parasite elemental composition will follow changes in the stoichiometry of its host, and that its reproductive success will decrease with host nutrient limitation. We tested this hypothesis by following the response of a host-parasite system to changes in nitrogen (N) and phosphorus (P) supply in a controlled laboratory experiment. To this end, we exposed a fungal parasite (the chytrid Rhizophydium megarrhizum) to its host (the freshwater cyanobacterium Planktothrix rubescens) under control, low N:P and high N:P conditions. Host N:P followed treatment conditions, with a decreased N:P ratio under low N:P supply, and an increased N:P ratio under high N:P supply, as compared to the control. Shifts in host N:P stoichiometry were reflected in the parasite stoichiometry. Furthermore, at low N:P supply, host intracellular microcystin concentration was lowered as compared to high N:P supply. In contrast to our hypothesis, zoospore production decreased at low N:P and increased at high N:P ratio as compared to the control. These findings suggest that fungal parasites have a relatively high N, but low P requirement. Furthermore, zoospore elemental content, and thereby presumably their size, decreased at high N:P ratios. From these results we hypothesize that fungal parasites may exhibit a trade-off between zoospore size and production. Since zooplankton can graze on chytrid zoospores, changes in parasite production, stoichiometry and cell size may have implications for aquatic food web dynamics.

Ecological resilience in lakes and the conjunction fallacy.

There is a pressing need to apply stability and resilience theory to environmental management to restore degraded ecosystems effectively and to mitigate the effects of impending environmental change. Lakes represent excellent model case studies in this respect and have been used widely to demonstrate theories of ecological stability and resilience that are needed to underpin preventative management approaches. However, we argue that this approach is not yet fully developed because the pursuit of empirical evidence to underpin such theoretically grounded management continues in the absence of an objective probability framework. This has blurred the lines between intuitive logic (based on the elementary principles of probability) and extensional logic (based on assumption and belief) in this field.

Temperature alters host genotype-specific susceptibility to chytrid infection.

The cost of parasitism often depends on environmental conditions and host identity. Therefore, variation in the biotic and abiotic environment can have repercussions on both, species-level host-parasite interaction patterns but also on host genotype-specific susceptibility to disease. We exposed seven genetically different but concurrent strains of the diatom Asterionella formosa to one genotype of its naturally co-occurring chytrid parasite Zygorhizidium planktonicum across five environmentally relevant temperatures. We found that the thermal tolerance range of the tested parasite genotype was narrower than that of its host, providing the host with a "cold" and "hot" thermal refuge of very low or no infection. Susceptibility to disease was host genotype-specific and varied with temperature level so that no genotype was most or least resistant across all temperatures. This suggests a role of thermal variation in the maintenance of diversity in disease related traits in this phytoplankton host. The duration and intensity of chytrid parasite pressure on host populations is likely to be affected by the projected changes in temperature patterns due to climate warming both through altering temperature dependent disease susceptibility of the host and, potentially, through en- or disabling thermal host refugia. This, in turn may affect the selective strength of the parasite on the genetic architecture of the host population.

Chytrid epidemics may increase genetic diversity of a diatom spring-bloom.

Contrary to expectation, populations of clonal organisms are often genetically highly diverse. In phytoplankton, this diversity is maintained throughout periods of high population growth (that is, blooms), even though competitive exclusion among genotypes should hypothetically lead to the dominance of a few superior genotypes. Genotype-specific parasitism may be one mechanism that helps maintain such high-genotypic diversity of clonal organisms. Here, we present a comparison of population genetic similarity by estimating the beta-dispersion among genotypes of early and peak bloom populations of the diatom Asterionella formosa for three spring-blooms under high or low parasite pressure. The Asterionella population showed greater beta-dispersion at peak bloom than early bloom in the 2 years with high parasite pressure, whereas the within group dispersion did not change under low parasite pressure. Our findings support that high prevalence parasitism can promote genetic diversification of natural populations of clonal hosts.

GENOTYPE-BY-TEMPERATURE INTERACTIONS MAY HELP TO MAINTAIN CLONAL DIVERSITY IN ASTERIONELLA FORMOSA (BACILLARIOPHYCEAE).

Marine and freshwater phytoplankton populations often show large clonal diversity, which is in disagreement with clonal selection of the most vigorous genotype(s). Temporal fluctuation in selection pressures in variable environments is a leading explanation for maintenance of such genetic diversity. To test the influence of temperature as a selection force in continually (seasonally) changing aquatic systems we carried out reaction norms experiments on co-occurring clonal genotypes of a ubiquitous diatom species, Asterionella formosa Hassall, across an environmentally relevant range of temperatures. We report within population genetic diversity and extensive diversity in genotype-specific reaction norms in growth rates and cell size traits. Our results showed genotype by environment interactions, indicating that no genotype could outgrow all others across all temperature environments. Subsequently, we constructed a model to simulate the relative proportion of each genotype in a hypothetical population based on genotype and temperature-specific population growth rates. This model was run with different seasonal temperature patterns. Our modeling exercise showed a succession of two to several genotypes becoming numerically dominant depending on the underlying temperature pattern. The results suggest that (temperature) context dependent fitness may contribute to the maintenance of genetic diversity in isolated populations of clonally reproducing microorganisms in temporally variable environments.