Foraging mechanisms in excavate flagellates shed light on the functional ecology of early eukaryotes.

The phagotrophic flagellates described as "typical excavates" have been hypothesized to be morphologically similar to the Last Eukaryotic Common Ancestor and understanding the functional ecology of excavates may therefore help shed light on the ecology of these early eukaryotes. Typical excavates are characterized by a posterior flagellum equipped with a vane that beats in a ventral groove. Here, we combined flow visualization and observations of prey capture in representatives of the three clades of excavates with computational fluid dynamic modeling, to understand the functional significance of this cell architecture. We record substantial differences amongst species in the orientation of the vane and the beat plane of the posterior flagellum. Clearance rate magnitudes estimated from flow visualization and modeling are both like that of other similarly sized flagellates. The interaction between a vaned flagellum beating in a confinement is modeled to produce a very efficient feeding current at low energy costs, irrespective of the beat plane and vane orientation and of all other morphological variations. Given this predicted uniformity of function, we suggest that the foraging systems of typical excavates studied here may be good proxies to understand those potentially used by our distant ancestors more than 1 billion years ago.

The fluid dynamics of barnacle feeding.

Sessile barnacles feed by sweeping their basket-like cirral fan through the water, intercepting suspended prey. A primary component of the diet of adult barnacles is copepods that are sensitive to fluid disturbances and capable of escaping. How do barnacles manage to capture copepods despite the fluid disturbances they generate? We examined this question by describing the feeding current architecture of 1 cm sized Balanus crenatus using particle image velocimetry, and by studying the trajectories of captured copepods and the escapes of evading copepods. We found that barnacles produce a feeding current that arrives both from behind and the sides of the barnacle. The flow from the sides represents quiescent corridors of low fluid deformation and uninterrupted by the beating cirral fan. Potential prey arriving from behind are likely to encounter the cirral fan and, hence, capture here is highly unlikely. Accordingly, most captured copepods arrived through the quiet corridors, while most copepods arriving from behind managed to escape. Thus, it is the unique feeding flow architecture that allows feeding on evasive prey. We used the Landau-Squire jet as a simple model of the feeding current. For the Reynolds number of our experiments, the model reproduces the main features of the feeding current, including the lateral feeding corridors. Furthermore, the model suggests that smaller barnacle specimens, operating at lower Reynolds numbers, will produce a fore-aft symmetric feeding current without the lateral corridors. This suggests an ontogenetic diet shift from non-evasive prey to inclusion of evasive prey as the barnacle grows.

The function of the feeding groove of 'typical excavate' flagellates.

Phagotrophic flagellates are the main consumers of bacteria and picophytoplankton. Despite their ecological significance in the 'microbial loop', many of their predation mechanisms remain unclear. 'Typical excavates' bear a ventral groove, where prey is captured for ingestion. The consequences of feeding through a 'semi-rigid' furrow on the prey size range have not been explored. An unidentified moving element called 'the wave' that sweeps along the bottom of the groove toward the site of phagocytosis has been observed in a few species; its function is unclear. We investigated the presence, behavior, and function of the wave in four species from the three excavate clades (Discoba, Metamonada, and Malawimonadida) and found it present in all studied cases, suggesting the potential homology of this feature across all three groups. The wave displayed a species-specific behavior and was crucial for phagocytosis. The morphology of the feeding groove had an upper-prey size limit for successful prey captures, but smaller particles were not constrained. Additionally, the ingestion efficiencies were species dependent. By jointly studying these feeding traits, we speculate on adaptations to differences in food availability to better understand their ecological functions.

Foraging trade-offs, flagellar arrangements, and flow architecture of planktonic protists.

Unicellular flagellated protists are a key element in aquatic microbial food webs. They all use flagella to swim and to generate feeding currents to encounter prey and enhance nutrient uptake. At the same time, the beating flagella create flow disturbances that attract flow-sensing predators. Protists have highly diverse flagellar arrangements in terms of number of flagella and their position, beat pattern, and kinematics, but it is unclear how the various arrangements optimize the fundamental trade-off between resource acquisition and predation risk. Here we describe the near-cell flow fields produced by 15 species and demonstrate consistent relationships between flagellar arrangement and swimming speed and between flagellar arrangement and flow architecture, and a trade-off between resource acquisition and predation risk. The flow fields fall in categories that are qualitatively described by simple point force models that include the drag force of the moving cell body and the propulsive forces of the flagella. The trade-off between resource acquisition and predation risk varies characteristically between flow architectures: Flagellates with multiple flagella have higher predation risk relative to their clearance rate compared to species with only one active flagellum, with the exception of the highly successful dinoflagellates that have simultaneously achieved high clearance rates and stealth behavior due to a unique flagellar arrangement. Microbial communities are shaped by trade-offs and environmental constraints, and a mechanistic explanation of foraging trade-offs is a vital part of understanding the eukaryotic communities that form the basis of pelagic food webs.

The effect of tethering on the clearance rate of suspension-feeding plankton.

Many planktonic suspension feeders are attached to particles or tethered by gravity when feeding. It is commonly accepted that the feeding flows of tethered suspension feeders are stronger than those of their freely swimming counterparts. However, recent flow simulations indicate the opposite, and the cause of the opposing conclusions is not clear. To explore the effect of tethering on suspension feeding, we use a low-Reynolds-number flow model. We find that it is favorable to be freely swimming instead of tethered since the resulting feeding flow past the cell body is stronger, leading to a higher clearance rate. Our result underscores the significance of the near-field flow in shaping planktonic feeding modes, and it suggests that organisms tether for reasons that are not directly fluid dynamical (e.g., to stay near surfaces where the concentration of bacterial prey is high).

Heterotrophic eukaryotes show a slow-fast continuum, not a gleaner-exploiter trade-off.

Gleaners and exploiters (opportunists) are organisms adapted to feeding in nutritionally poor and rich environments, respectively. A trade-off between these two strategies-a negative relationship between the rate at which organisms can acquire food and ingest it-is a critical assumption in many ecological models. Here, we evaluate evidence for this trade-off across a wide range of heterotrophic eukaryotes from unicellular nanoflagellates to large mammals belonging to both aquatic and terrestrial realms. Using data on the resource acquisition and ingestion rates in >500 species, we find no evidence of a trade-off across species. Instead, there is a positive relationship between maximum clearance rate and maximum ingestion rate. The positive relationship is not a result of lumping together diverse taxa; it holds within all subgroups of organisms we examined as well. Correcting for differences in body mass weakens but does not reverse the positive relationship, so this is not an artifact of size scaling either. Instead, this positive relationship represents a slow-fast gradient in the "pace of life" that overrides the expected gleaner-exploiter trade-off. Other trade-offs must therefore shape ecological processes, and investigating them may provide deeper insights into coexistence, competitive dynamics, and biodiversity patterns in nature. A plausible target for study is the well-documented trade-off between growth rate and predation avoidance, which can also drive the slow-fast gradient we observe here.

Evolution of toxins as a public good in phytoplankton.

Toxic phytoplankton blooms have increased in many waterbodies worldwide with well-known negative impacts on human health, fisheries and ecosystems. However, why and how phytoplankton evolved toxin production is still a puzzling question, given that the producer that pays the costs often shares the benefit with other competing algae and thus provides toxins as a 'public good' (e.g. damaging a common competitor or predator). Furthermore, blooming phytoplankton species often show a high intraspecific variation in toxicity and we lack an understanding of what drives the dynamics of coexisting toxic and non-toxic genotypes. Here, by using an individual-based two-dimensional model, we show that small-scale patchiness of phytoplankton strains caused by demography can explain toxin evolution in phytoplankton with low motility and the maintenance of genetic diversity within their blooms. This patchiness vanishes for phytoplankton with high diffusive motility, suggesting different evolutionary pathways for different phytoplankton groups. In conclusion, our study reveals that small-scale spatial heterogeneity, generated by cell division and counteracted by diffusive cell motility and turbulence, can crucially affect toxin evolution and eco-evolutionary dynamics in toxic phytoplankton species. This contributes to a better understanding of conditions favouring toxin production and the evolution of public goods in asexually reproducing organisms in general.

Costs and benefits of predator-induced defence in a toxic diatom.

Phytoplankton employ a variety of defence mechanisms against predation, including production of toxins. Domoic acid (DA) production by the diatom Pseudo-nitzschia spp. is induced by the presence of predators and is considered to provide defence benefits, but the evidence is circumstantial. We exposed eight different strains of P. seriata to chemical cues from copepods and examined the costs and the benefits of toxin production. The magnitude of the induced toxin response was highly variable among strains, while the costs in terms of growth reduction per DA cell quota were similar and the trade-off thus consistent. We found two components of the defence in induced cells: (i) a 'private good' in terms of elevated rejection of captured cells and (ii) a 'public good' facilitated by a reduction in copepod feeding activity. Induced cells were more frequently rejected by copepods and rejections were directly correlated with DA cell quota and independent of access to other food items. By contrast, the public-good effect was diminished by the presence of alternative prey suggesting that it does not play a major role in bloom formation and that its evolution is closely associated with the grazing-deterrent private good.

Machine learning techniques to characterize functional traits of plankton from image data.

Plankton imaging systems supported by automated classification and analysis have improved ecologists' ability to observe aquatic ecosystems. Today, we are on the cusp of reliably tracking plankton populations with a suite of lab-based and in situ tools, collecting imaging data at unprecedentedly fine spatial and temporal scales. But these data have potential well beyond examining the abundances of different taxa; the individual images themselves contain a wealth of information on functional traits. Here, we outline traits that could be measured from image data, suggest machine learning and computer vision approaches to extract functional trait information from the images, and discuss promising avenues for novel studies. The approaches we discuss are data agnostic and are broadly applicable to imagery of other aquatic or terrestrial organisms.

Hydrodynamics of microbial filter feeding.

Microbial filter feeders are an important group of grazers, significant to the microbial loop, aquatic food webs, and biogeochemical cycling. Our understanding of microbial filter feeding is poor, and, importantly, it is unknown what force microbial filter feeders must generate to process adequate amounts of water. Also, the trade-off in the filter spacing remains unexplored, despite its simple formulation: A filter too coarse will allow suitably sized prey to pass unintercepted, whereas a filter too fine will cause strong flow resistance. We quantify the feeding flow of the filter-feeding choanoflagellate Diaphanoeca grandis using particle tracking, and demonstrate that the current understanding of microbial filter feeding is inconsistent with computational fluid dynamics (CFD) and analytical estimates. Both approaches underestimate observed filtration rates by more than an order of magnitude; the beating flagellum is simply unable to draw enough water through the fine filter. We find similar discrepancies for other choanoflagellate species, highlighting an apparent paradox. Our observations motivate us to suggest a radically different filtration mechanism that requires a flagellar vane (sheet), something notoriously difficult to visualize but sporadically observed in the related choanocytes (sponges). A CFD model with a flagellar vane correctly predicts the filtration rate of D. grandis, and using a simple model we can account for the filtration rates of other microbial filter feeders. We finally predict how optimum filter mesh size increases with cell size in microbial filter feeders, a prediction that accords very well with observations. We expect our results to be of significance for small-scale biophysics and trait-based ecological modeling.

Nutrient affinity, half-saturation constants and the cost of toxin production in dinoflagellates.

The two parameters of the Michaelis-Menten model, the maximum uptake rate and the half-saturation constant, are not stochastically independent, and the half-saturation constant is not a measure of nutrient affinity, as commonly assumed. Failure to realise their interdependence and mechanistic interpretation may lead to the emergence of false trade-offs.

Dense Dwarfs versus Gelatinous Giants: The Trade-Offs and Physiological Limits Determining the Body Plan of Planktonic Filter Feeders.

Most marine plankton have a high energy (carbon) density, but some are gelatinous with approximately 100 times more watery bodies. How do those distinctly different body plans emerge, and what are the trade-offs? We address this question by modeling the energy budget of planktonic filter feeders across life-forms, from micron-sized unicellular microbes such as choanoflagellates to centimeter-sized gelatinous tunicates such as salps. We find two equally successful strategies, one being small with high energy density (dense dwarf) and the other being large with low energy density (gelatinous giant). The constraint that forces large-but not small-filter feeders to be gelatinous is identified as a lower limit to the size-specific filter area, below which the energy costs lead to starvation. A further limit is found from the maximum size-specific motor force that restricts the access to optimum strategies. The quantified constraints are discussed in the context of other resource-acquisition strategies. We argue that interception feeding strategies can be accessed by large organisms only if they are gelatinous. On the other hand, organisms that use remote prey sensing do not need to be gelatinous, even if they are large.

Trophic interactions drive the emergence of diel vertical migration patterns: a game-theoretic model of copepod communities.

Diel vertical migration (DVM), the daily movement of organisms through oceanic water columns, is mainly driven by spatio-temporal variations in the light affecting the intensity of predator-prey interactions. Migration patterns of an organism are intrinsically linked to the distribution of its conspecifics, its prey and its predators, each with their own fitness-seeking imperatives. We present a mechanistic, trait-based model of DVM for the different components of a pelagic community. Specifically, we consider size, sensory mode and feeding mode as key traits, representing a community of copepods that prey on each other and are, in turn, preyed upon by fish. Using game-theoretic principles, we explore the optimal distribution of the main groups of a planktonic pelagic food web simultaneously. Within one single framework, our model reproduces a whole suite of observed patterns, such as size-dependent DVM patterns of copepods and reverse migrations. These patterns can only be reproduced when different trophic levels are considered at the same time. This study facilitates a quantitative understanding of the drivers of DVM, and is an important step towards mechanistically underpinned predictions of DVM patterns and biologically mediated carbon export.

Bottom-up behaviourally mediated trophic cascades in plankton food webs.

Our traditional view of the interactions between marine organisms is conceptualized as food webs where species interact with one another mainly via direct consumption. However, recent research suggests that understudied non-consumptive interactions, such as behaviourally mediated indirect interactions (BMIIs), can influence marine ecosystems as much as consumptive effects. Here, we show, to our knowledge, the first experimental evidence and quantification of bottom-up BMIIs in plankton food webs. We used observational, modelling and experimental approaches to investigate how behavioural responses to resource availability influence predation mortality on grazers with different foraging strategies (ambushing versus active foraging). A three-level food chain was used: phytoplankton as resource, copepod nauplii as grazers of phytoplankton and a large copepod as a predator. Ambushers showed little change in foraging activity with resource availability, whereas active foragers decreased their foraging activity with increasing resources, which led to a decrease (24-50%) in predation mortality. Therefore, an increase in resources ('initiator') causes behavioural changes in active grazers ('transmitter'), which ultimately negatively affects predator ('receiver') consumption rates. Consequently, increase in resource abundance may result in decreasing energy transfer to higher trophic levels. These results indicate that behaviourally mediated interactions drive marine food web dynamics differently from that predicted by only density-mediated or consumptive interactions.

Silicified cell walls as a defensive trait in diatoms.

Diatoms contribute nearly half of the marine primary production. These microalgae differ from other phytoplankton groups in having a silicified cell wall, which is the strongest known biological material relative to its density. While it has been suggested that a siliceous wall may have evolved as a mechanical protection against grazing, empirical evidence of its defensive role is limited. Here, we experimentally demonstrate that grazing by adult copepods and nauplii on diatoms is approximately inversely proportional to their silica content, both within and among diatom species. While a sixfold increase in silica content leads to a fourfold decrease in copepod grazing, silicification provides no protection against protozoan grazers that directly engulf their prey. We also found that the wall provides limited protection to cells ingested by copepods, since less than 1% of consumed cells were alive in the faecal pellets. Moreover, silica deposition in diatoms decreases with increasing growth rates, suggesting a possible cost of defence. Overall, our results demonstrate that thickening of silica walls is an effective defence strategy against copepods. This suggests that the plasticity of silicification in diatoms may have evolved as a response to copepod grazing pressure, whose specialized tools to break silicified walls have coevolved with diatoms.

Toxic dinoflagellates produce true grazer deterrents.

Many phytoplankton species produce toxic substances, but their functional role is unclear. Specifically, it remains uncertain whether these compounds have a toxic or deterrent effect on grazers; only, the latter is consistent with toxins as defensive tools. Here, we show that 10 of 12 species or strains of toxic dinoflagellates were consumed at lower rates than a similarly sized nontoxic dinoflagellate by a copepod. Through video observations of individual prey-grazer interactions, we further demonstrate that the dominating mechanism is through capture, examination, and subsequent rejection of vital cells, that is, a true deterrent effect that offers a straightforward explanation to its evolution. We argue that the diversity of grazer responses to toxic phytoplankton reported in the literature, including toxic effects, and the high diversity of toxin profiles between strains of the same phytoplankton species reflect different stages of an ever-ongoing evolutionary arms race, facilitated by rapid adaptation of grazers to toxic substances. We further argue that defensive toxicity requires a chemical signal exterior to the cell that informs the grazer about the toxicity of the cell. The signal can be the toxin itself or just an aposematic signal of toxicity. In the former case, allelochemical effects may emerge at high cell concentrations as a nonadaptive side effect of a predator defenses.

Selection for life-history traits to maximize population growth in an invasive marine species.

Species establishing outside their natural range, negatively impacting local ecosystems, are of increasing global concern. They often display life-history features characteristic for r-selected populations with fast growth and high reproduction rates to achieve positive population growth rates (r) in invaded habitats. Here, we demonstrate substantially earlier maturation at a 2 orders of magnitude lower body mass at first reproduction in invasive compared to native populations of the comb jelly Mnemiopsis leidyi. Empirical results are corroborated by a theoretical model for competing life-history traits that predicts maturation at the smallest possible size to optimize r, while individual lifetime reproductive success (R0 ), optimized in native populations, is near constant over a large range of intermediate maturation sizes. We suggest that high variability in reproductive tactics in native populations is an underappreciated determinant of invasiveness, acting as substrate upon which selection can act during the invasion process.

Swim and fly: escape strategy in neustonic and planktonic copepods.

Copepods can respond to predators by powerful escape jumps that in some surface-dwelling forms may propel the copepod out of the water. We studied the kinematics and energetics of submerged and out-of-water jumps of two neustonic pontellid copepods, Anomalocera patersoni and Pontella mediterranea, and one pelagic calanoid copepod, Calanus helgolandicus (euxinus). We show that jumping out of the water does not happen just by inertia gained during the copepod's acceleration underwater, but also requires the force generated by the thoracic limbs when breaking through the water's surface to overcome surface tension, drag and gravity. The timing of this appears to be necessary for success. At the moment of breaking the water interface, the instantaneous velocity of the two pontellids reached 125 cm s-1, while their maximum underwater speed (115 cm s-1) was close to that of similarly sized C. helgolandicus (106 cm s-1). The average specific power produced by the two pontellids during out-of-water jumps (1700-3300 W kg-1 muscle mass) was close to that during submerged jumps (900-1600 W kg-1 muscle mass) and, in turn, similar to that produced during submerged jumps of C. helgolandicus (1300 W kg-1 muscle mass). The pontellids may shake off water adhering to their body by repeated strokes of the limbs during flight, which leads to a slight acceleration in the air. Our observations suggest that out-of-water jumps of pontellids are not dependent on any exceptional ability to perform this behavior but have the same energetic cost and are based on the same kinematic patterns and contractive capabilities of muscles as those of copepods swimming submerged.