Mixotrophic Nanoflagellates Research Articles

Disentangling the role of light and nutrient limitation on bacterivory by mixotrophic nanoflagellates.

Many phytoplankton taxa function on multiple trophic levels by combining photosynthesis and ingestion of bacteria, termed mixotrophy. Despite the recognition of mixotrophy as a universal functional trait, we have yet to fully resolve how environmental conditions influence community grazing rates in situ. A microcosm study was used to assess bacterivory by mixotrophic nanoflagellates following nutrient enrichment and light attenuation in a temperate lake. We found contrasting results based on assessment of mixotroph abundance or bacterivory. Despite an interactive effect of nutrient enrichment and light attenuation on mixotroph abundance, significant differences within light treatments were observed only after enrichment with P or N + P. The greatest abundance of mixotrophs across treatments occurred under co-nutrient enrichment with full exposure to irradiance. However, bacterivory by mixotrophic nanoflagellates was greatest under shaded conditions after either N or P enrichment. We suggest that PAR availability dampened the stimulatory effect of nutrient limitation, and bacterivory supplemented a suboptimal photosynthetic environment. In a saturating light regime, the mixotrophic community was less driven to ingest bacteria because photosynthesis was able to satisfy energetic demands. These findings quantify community bacterivory in response to environmental drivers that may characterize future ecosystem conditions and highlight the importance of considering grazing rates in conjunction with abundance of mixotrophic protists.

Evidence for evolutionary adaptation of mixotrophic nanoflagellates to warmer temperatures.

Mixotrophs, organisms that combine photosynthesis and heterotrophy to gain energy, play an important role in global biogeochemical cycles. Metabolic theory predicts that mixotrophs will become more heterotrophic with rising temperatures, potentially creating a positive feedback loop that accelerates carbon dioxide accumulation in the atmosphere. Studies testing this theory have focused on phenotypically plastic (short-term, non-evolutionary) thermal responses of mixotrophs. However, as small organisms with short generation times and large population sizes, mixotrophs may rapidly evolve in response to climate change. Here, we present data from a 3-year experiment quantifying the evolutionary response of two mixotrophic nanoflagellates to temperature. We found evidence for adaptive evolution (increased growth rates in evolved relative to acclimated lineages) in the obligately phototrophic strain, but not in the facultative phototroph. All lineages showed trends of increased carbon use efficiency, flattening of thermal reaction norms, and a return to homeostatic gene expression. Generally, mixotrophs evolved reduced photosynthesis and higher grazing with increased temperatures, suggesting that evolution may act to exacerbate mixotrophs' effects on global carbon cycling.

Annual Cycle of the Synechococcus spp. and Picoeukaryotic Growth and Loss Rates in a Subtropical Coastal Ecosystem

Seasonal variations in the picophytoplankton community structure (Synechococcus spp. and picoeukaryotes) were studied by flow cytometry in the coastal ecosystem of the subtropical western Pacific from October 2019 to September 2020. Synechococcus spp. was dominant in abundance during the study period, with its density ranging from 0.05 to 5.6 × 104 cells mL−1; its maximum occurred in July 2020. Picoeukaryotes were less abundant, with their density ranging from 0.2 to 13.6 × 103 cells mL−1. Their highest abundance was recorded in January 2020. The growth rates of Synechococcus spp. and picoeukaryotes ranged from −0.39 to 1.42 d−1 and 0.38 to 2.46 d−1, respectively, throughout the study period. Overall, the growth rate of the picoeukaryotes was significantly higher than that of Synechococcus spp. It is interesting to note that the grazing mortality of Synechococcus spp. and picoeukaryotes during the warmer period (April to September) was relatively low. Based on this study, we suggest that mixotrophic nanoflagellates lowered their feeding activity that obtained nutrients from prey and instead used additional nutrients during the incubation experiments. Our study demonstrated that a shift in the picophytoplankton community composition and grazing activity of predacious nanoflagellates in cold and warm periods can impact on the seasonal dynamics of the microbial food web.

Response of Growth and Grazing Rate of Nanoflagellates on Synechococcus spp. to Experimental Nutrient Enrichment

As important bacterivores in planktonic food webs, mixotrophic nanoflagellates cancause mortality in marine Synechococcus spp. Our previous study found that the pigmented nanoflagellate (PNF) has a significant grazing impact on Synechococcus spp. In the current study, we applied the dilution approach to test the growth and grazing rates of nanoflagellates on Synechococcus spp. We then compared the differences between experimental nutrient additions and in situ conditions in the coastal waters of the East China Sea during the summer season from July to September. The growth rates of Synechococcus spp. in the ambient environment were between 0.54 and 0.62 day−1, which were slightly higher than the 0.56 and 0.66 day−1 with nutrient enrichment in summer. In contrast, our nutrient enrichment experiments produced a marked decline approximately from 21% to 58% in the nanoflagellate grazing rate on Synechococcus spp. The reason was that the mixotrophic PNFs directly used the added nutrients and reduced their supply of nutrients from prey during the incubation experiments.

Toxins and Other Bioactive Metabolites in Deep Chlorophyll Layers Containing the Cyanobacteria Planktothrix cf. isothrix in Two Georgian Bay Embayments, Lake Huron.

The understanding of deep chlorophyll layers (DCLs) in the Great Lakes—largely reported as a mix of picoplankton and mixotrophic nanoflagellates—is predominantly based on studies of deep (>30 m), offshore locations. Here, we document and characterize nearshore DCLs from two meso-oligotrophic embayments, Twelve Mile Bay (TMB) and South Bay (SB), along eastern Georgian Bay, Lake Huron (Ontario, Canada) in 2014, 2015, and 2018. Both embayments showed the annual formation of DCLs, present as dense, thin, metalimnetic plates dominated by the large, potentially toxic, and bloom-forming cyanobacteria Planktothrix cf. isothrix. The contribution of P. cf. isothrix to the deep-living total biomass (TB) increased as thermal stratification progressed over the ice-free season, reaching 40% in TMB (0.6 mg/L at 9.5 m) and 65% in South Bay (3.5 mg/L at 7.5 m) in 2015. The euphotic zone in each embayment extended down past the mixed layer, into the nutrient-enriched hypoxic hypolimnia, consistent with other studies of similar systems with DCLs. The co-occurrence of the metal-oxidizing bacteria Leptothrix spp. and bactivorous flagellates within the metalimnetic DCLs suggests that the microbial loop plays an important role in recycling nutrients within these layers, particularly phosphate (PO4) and iron (Fe). Samples taken through the water column in both embayments showed measurable concentrations of the cyanobacterial toxins microcystins (max. 0.4 µg/L) and the other bioactive metabolites anabaenopeptins (max. ~7 µg/L) and cyanopeptolins (max. 1 ng/L), along with the corresponding genes (max. in 2018). These oligopeptides are known to act as metabolic inhibitors (e.g., in chemical defence against grazers, parasites) and allow a competitive advantage. In TMB, the 2018 peaks in these oligopeptides and genes coincided with the P. cf. isothrix DCLs, suggesting this species as the main source. Our data indicate that intersecting physicochemical gradients of light and nutrient-enriched hypoxic hypolimnia are key factors in supporting DCLs in TMB and SB. Microbial activity and allelopathy may also influence DCL community structure and function, and require further investigation, particularly related to the dominance of potentially toxigenic species such as P. cf. isothrix.

Lake Ecosystem Robustness and Resilience Inferred from a Climate-Stressed Protistan Plankton Network.

Network analyses of biological communities allow for identifying potential consequences of climate change on the resilience of ecosystems and their robustness to resist stressors. Using DNA metabarcoding datasets from a three-year-sampling (73 samples), we constructed the protistan plankton co-occurrence network of Lake Zurich, a model lake ecosystem subjected to climate change. Despite several documentations of dramatic lake warming in Lake Zurich, our study provides an unprecedented perspective by linking changes in biotic association patterns to climate stress. Water temperature belonged to the strongest environmental parameters splitting the data into two distinct seasonal networks (October–April; May–September). The expected ecological niche of phytoplankton, weakened through nutrient depletion because of permanent thermal stratification and through parasitic fungi, was occupied by the cyanobacterium Planktothrix rubescens and mixotrophic nanoflagellates. Instead of phytoplankton, bacteria and nanoflagellates were the main prey organisms associated with key predators (ciliates), which contrasts traditional views of biological associations in lake plankton. In a species extinction scenario, the warm season network emerged as more vulnerable than the cold season network, indicating a time-lagged effect of warmer winter temperatures on the communities. We conclude that climate stressors compromise lake ecosystem robustness and resilience through species replacement, richness differences, and succession as indicated by key network properties.

Role of mixotrophic nanoflagellates in the Eastern Mediterranean microbial food web

In the oligotrophic, phosphorus (P)-limited Eastern Mediterranean Sea (EMS), grazing on heterotrophic bacteria (HB) by pigmented nanoflagellates (PNF) through mixotrophy is a significant source of bacterial mortality and is P-dependent. Heterotrophic nanoflagellates (HNF) are also important consumers of HB. However, there is still no conceptual framework describing the flows of carbon (C) and P through the EMS microbial food web that takes into account the mixotrophic behavior of PNF. In the present modelling study, we explore qualitatively the pathways of C- and P-flow through the HB-mixotrophic PNF-HNF link under varying availability of inorganic P. At low P availability, given the assumption that PNF can fulfill most of their P requirements through HB consumption when P-limited, model simulations show that PNF support bacterial production and P uptake by supplying HB with dissolved organic carbon (DOC), leading to a win-win stable scenario, with PNF holding a key role in P-transfer through the microbial food web. Conversely, increased P availability results in lower PNF grazing upon HB, amplifying the relative contribution of HNF to bacterial grazing. Based on our modelling study we propose a novel conceptual framework for bacteria-nanoflagellate interactions in the EMS, where PNF have the functional role of DOC primary producers and also share with HNF the functional role of bacterial consumers via mixotrophy. This alternative microbial loop operates in a Pdependent way. Our modelling study pinpoints the significance of the adaptive physiology of PNF for the functioning of the oligotrophic ecosystem of the EMS.

Plasticity in the grazing ecophysiology of Florenciella (Dichtyochophyceae), a mixotrophic nanoflagellate that consumes Prochlorococcus and other bacteria

AbstractMixotrophic nanoflagellates can account for more than half of the bacterivory in the sunlit ocean, yet very little is known about their ecophysiology. Here, we characterize the grazing ecology of an open‐ocean mixotroph in the genus Florenciella (class Dictyochophyceae). Members of this class were indirectly implicated as major consumers of Prochlorococcus and Synechococcus in the oligotrophic North Pacific Subtropical Gyre, but their phagotrophic capabilities have never been investigated. Our studies showed that Florenciella readily consumed Prochlorococcus, Synechococcus, and heterotrophic bacteria, and that the ingested prey relieved nutrient limitations on growth. Florenciella grew faster (3 d−1) in nitrogen‐deplete medium given sufficient live Synechococcus, than in nitrogen‐replete K medium (2 d−1), but it did not grow in continuous darkness. Grazing rates were substantially higher under nutrient limitation and showed a hint of diel variability, with rates tending to be highest near the end of the light period. An apparent trade‐off between the maximum clearance rate (5 nL Florenciella−1 h−1) and the maximum ingestion rate (up to ∼ 10 prey cells Florenciella−1 h−1) across experiments suggests that grazing behavior may also vary in response to prey concentration. If the observed grazing rates are representative of other open‐ocean mixotrophs, their collective activity could account for a significant fraction of the daily cyanobacterial mortality. This study provides essential parameters for understanding the grazing ecology of a common marine mixotroph and the first characterization of mixotrophic nanoflagellate functional responses when feeding on unicellular cyanobacteria, the dominant marine primary producers in the oligotrophic ocean.

Modelling the consequence of glacier retreat on mixotrophic nanoflagellate bacterivory: a Bayesian approach

Mixotrophic nanoflagellate bacterivory is affected by light; however, in glacially influenced lakes, glacial clay may also interfere with prey uptake. Mechanistic models based on prior quantitative hypotheses and tested with field data are useful for predicting these predator–prey interactions under a changing climate. We modelled the effect of glacial particles on the bacterivory of mixotrophic nanoflagellates, the dominant phytoplankton in mountain lakes in the North Patagonian Andes. Our model equation is based on the classical Michaelis–Menten formulation and Platt's photosynthesis–irradiance curve to account for the interference by glacial particles and the effect of light intensity, respectively. Bayesian inference was applied to estimate model parameters using data from field bacterivory experiments. Light experiments (LE) were performed in lakes with different levels of light penetration without clay particles, while clay experiments (CE) were performed with an experimental gradient of glacial clay concentration at constant light intensity. The LE showed a non‐monotonic effect of light intensity and the CE revealed an exponential negative effect of clay concentration. The obtained model was tested with two independent experiments carried out in a turbid proglacial lake. Our proposed mechanistic model successfully predicted the combined effect of light availability and particle interference on the mixotrophic nanoflagellate–bacteria relationship. The model demonstrated that the effect of light is dampened as clay concentration increases.

Modelling the nutritional strategies in mixotrophic nanoflagellates

Mixotrophic nanoflagellates (MNF) play a key role in the carbon flux within aquatic ecosystems. They contribute significantly to primary production while, at the same time, they are important consumers of bacteria. MNF comprise a diverse assemblage with a whole range of photosynthetic and feeding potential and with various metabolic responses to environmental drivers, such as light, nutrients and prey availability. However, in most cases MNF are modelled as a single functional group with no reference to the distinct nutritional strategies and constraints of the different types of MNF. Here we develop a mathematical framework for the distinct functional responses of four types of MNF. Our approach is based on the Dynamic Energy Budget (DEB) theory and the concept of Synthesing Unit (SU), which is used to merge phototrophic and phagotrophic nutrition into mixotrophic growth. The resulting models describe explicitly the functional diversity of the MNF group taking into account the dynamic interaction of phototrophy and phagotrophy within the four types of MNF. Our results are in agreement with theoretical expectations and a wide range of experimental observations for the different types of MNF. Our simulations suggest that the growth dynamics of the four types of MNF are affected differently by the availability of resources. Moreover, using our models to compare the rates of organic carbon production and prey consumption by the various MNF types, we show that their net ecosystem role, as producers or consumers, depends on the mixotrophic strategy and it can vary as a function of the prevailing environmental conditions. Overall, our study highlights the importance of mechanistically modelling the distinct metabolic responses of the different types of MNF to environmental drivers in order to better understand their role in the carbon cycle in anticipation of global environmental changes.

Abiotic and Biotic Factors Affecting the Ingestion Rates of Mixotrophic Nanoflagellates (Haptophyta)

Mixotrophic haptophytes comprise one of several important groups of mixotrophic nanoflagellates in the pelagic environment. This study aimed to investigate if phagotrophy in mixotrophic haptophytes is regulated by light or other factors in the surface (SE) and bottom (BE) of the euphotic zone in the subtropical northwestern Pacific Ocean. We estimated the rates of bacterial ingestion by haptophytes using fluorescently labeled bacteria (FLBs) and fluorescence in situ hybridization. Haptophyte diversity and abundance were also investigated in the same sampling area. The annual mean abundance of haptophytes was 419 ± 85.6cellsmL-1 in both SE and BE. Cells 3-5μm in size were the dominant group in all haptophytes and accounted for majority of bacteria standing stock removed by haptophytes (53%). Most haptophyte ingestion rates (IRs) were not significantly different between the two layers (average SE ingestion rate: 12.5 ± 2.29bacHap-1h-1; BE: 14.7 ± 3.03bacHap-1h-1). Furthermore, the haptophyte IRs were negatively correlated with nitrate concentrations in the SE and positively correlated with bacterial abundances in the BE, which accounts for the significantly high IRs in August 2012 and 2013. These findings imply that mixotrophic haptophytes in this region had different factors affecting phagotrophy to adapt to the ambient light intensity alterations between SE and BE.

Vulnerability of mixotrophic algae to nutrient pulses and UVR in an oligotrophic Southern and Northern Hemisphere lake

Nutrient inputs and ultraviolet radiation (UVR) are global factors affecting the structure and functioning of aquatic ecosystems, particularly clear-water ecosystems. We performed experiments in two model lakes highly exposed to UVR fluxes in order to test the effect that future increases in mineral nutrients transported by dust aerosol might exert on primary producers depending on the likelihood of atmospheric inputs. Lake La Caldera (Northern Hemisphere) has been receiving recurrent dust inputs from the Sahara Desert while lake Los Cántaros (Southern Hemisphere) has been less affected by dust aerosol. UVR × Nutrient synergistically stimulated primary production (PP), chlorophyll a (Chl a), with a smaller increase in phytoplanktonic biomass in La Caldera, but not in Los Cántaros, where nutrient addition unmasked the UVR inhibitory effect on phytoplankton. The proportional decrease of mixotrophic nanoflagellates (MNFs) after the nutrient pulse (in Los Cántaros) and the long-term decline of MNFs in La Caldera associated with the increase in aerosol-dust intrusions from the Sahara during the last 40 years suggest that a future scenario of intensified aerosol events from desert and desertified areas would not only reduce functional diversity with the decline of MNFs, but would ultimately alter the C flux towards the grazing chain in oligotrophic ecosystems.

Rising nutrient-pulse frequency and high UVR strengthen microbial interactions

Solar radiation and nutrient pulses regulate the ecosystem’s functioning. However, little is known about how a greater frequency of pulsed nutrients under high ultraviolet radiation (UVR) levels, as expected in the near future, could alter the responses and interaction between primary producers and decomposers. In this report, we demonstrate through a mesocosm study in lake La Caldera (Spain) that a repeated (press) compared to a one-time (pulse) schedule under UVR prompted higher increases in primary (PP) than in bacterial production (BP) coupled with a replacement of photoautotrophs by mixotrophic nanoflagellates (MNFs). The mechanism underlying these amplified phytoplanktonic responses was a dual control by MNFs on bacteria through the excretion of organic carbon and an increased top-down control by bacterivory. We also show across a 6-year whole-lake study that the changes from photoautotrophs to MNFs were related mainly to the frequency of pulsed nutrients (e.g. desert dust inputs). Our results underscore how an improved understanding of the interaction between chronic and stochastic environmental factors is critical for predicting ongoing changes in ecosystem functioning and its responses to climatically driven changes.

Heterotrophic and mixotrophic nanoflagellates in a mesotrophic lake: Abundance and grazing impacts across season and depth

AbstractNanoflagellates are recognized as fundamental members of the planktonic microbial food web due to their contribution to photosynthetic fixation of carbon and phagotrophic ingestion of bacteria. Although the presence, and often dominance, of mixotrophic nutrition among phytoflagellates has been well documented within a variety of aquatic ecosystems and in a range of taxa, few studies have assessed the abundance and grazing impact of mixotrophs in comparison to their photosynthetic and heterotrophic counterparts across seasons at multiple depths. Abundance and, as appropriate, bacterivory of phototrophic, heterotrophic, and mixotrophic nanoflagellates were quantified at three depths over a 15‐month period in Lake Lacawac (Pocono Mountains, Pennsylvania, U.S.A.) via microscopic analysis of ingested bacterial surrogates. The absolute and relative abundances of all nanoflagellate trophic groups varied with season and depth, likely as a result of differences in physiological tolerance. Whereas the abundance of phototrophic and heterotrophic nanoflagellates varied with depth in both the presence and absence of thermal stratification, mixotrophic protists were evenly distributed throughout the water column during mixis. The abundances of phototrophic and mixotrophic protists were significantly correlated, but only in surface waters during periods of thermal stratification. Grazing rate and consequent impact by bacterivorous nanoflagellates varied with depth across season, particularly during thermal stratification. Mixotrophic bacterivory exceeded that of heterotrophs in the epi‐ and metalimnion during stratification, removing an estimated maximum of 80% of the bacterial standing stock d−1. The relative contribution of heterotrophic nanoflagellates to community bacterivory increased with depth, even during mixis, and dominated the grazing impact in the hypolimnion.

Distribution of mixotrophic nanoflagellates along the latitudinal transect of the central North Pacific

To elucidate the ecological importance of mixotrophic nanoflagellates in the open ocean and the environmental factors that regulate their abundance, we surveyed latitudinal distributions of autotrophic, mixotrophic and heterotrophic nanoflagellates in the central North Pacific Ocean along a transect at 170°W. Mixotrophic nanoflagellates significantly contributed (26–64 %) to total bacterivory, as measured by the fluorescently-labeled bacteria method, from the equatorial through the subarctic regions, which reinforces the importance of mixotrophic nanoflagellates as a trophic link in the open ocean. The proportion of mixotrophic to total plastidic nanoflagellates was significantly higher in the nutrient-depleted subtropical gyre than in other regions, sometimes exceeding 10 %. Additionally, the proportion was negatively correlated with soluble reactive phosphorus concentration within the tropical and subtropical waters, suggesting that low nutrient availability could facilitate phagotrophy of plastidic nanoflagellates, which may explain the survival of nano-sized eukaryotic phytoplankton in the ultraoligotrophic water. In the subarctic regions, the proportion exhibited no obvious relationship with any environmental parameter. Conversely, the numerical proportion of mixotrophic nanoflagellates in total phagotrophic nanoflagellates (sum of mixotrophic and heterotrophic nanoflagellates) was positively correlated with nutrient concentrations. In contrast to macronutrient availability, the physical stability of the water column did not appear to affect the contribution of mixotrophic nanoflagellates.

Vertical distribution of pigmented and non-pigmented nanoflagellates in the East China Sea

Nanoflagellates can be separated into two groups according to their trophic mode, i.e. pigmented nanoflagellates (PNF) and heterotrophic nanoflagellates (HNF). However, a newly identified group, mixotrophic nanoflagellates (MNF), are pigmented and show the ability of prey on bacteria. To examine the vertical variations in PNF and HNF abundances, as well as their relationships and the nutritional strategies that they might use, two summer cruises were undertaken in the East China Sea in July 2011 (OR1 966) and July 2012 (OR1 1004). The results show that both HNF and PNF abundances decline with increasing water depth. Vertical variations of abundances are believed to be influenced by prey and light, for HNF and PNF respectively. Over a large part of the sampling area, the ratio of PNF to HNF abundances is about 1:1 in the disphotic and euphotic zones, but exceeds 1.5 in the nutrient-depleted environment along the margin of the continental shelf. The correlation between PNF abundance and bacteria/Synechococcus abundance is positive where PNF/HNF >1.5. However, there is no significant correlation between PNF/HNF abundance when PNF/HNF >1.5 and light/nutrients, indicating that vertical distributions are influenced mainly by prey (bacteria and Synechococcus) in the nutrient-depleted environment. This study assumes that PNF consists mostly of MNF. In the euphotic zone they receive energy from photosynthesis, which is stimulated by the available nutrients from grazing. Their abundance is thus higher than that of HNF. However, in the disphotic zone, both PNF and HNF satisfy their nutrient demands by grazing, and PNF/HNF is close to 1. In other words, mixotrophy might be the main trophic mode for PNF in the nutrient-depleted, oligotrophic environment. Meanwhile, in deeper water (300m), the much lower prey density means that MNF cannot satisfy the basic energy demands of metabolism and photosynthesis, and thus HNF abundance exceeds that of PNF.

Grazing-induced Synechococcus microcolony formation: experimental insights from two freshwater phylotypes.

Freshwater cyanobacteria of the genus Synechococcus are ubiquitous and organized either as single cells of diverse morphology or as microcolonies of different size. We studied the formation of microcolonies induced by the mixotrophic nanoflagellate Poterioochromonas sp. grazing on two Synechococcus strains belonging to phylotypes with different content of phycobiliproteins (PE: phycoerythrin-rich cells, L.Albano Group A; PC: phycocyanin-rich cells, MW101C3 Group I). The quantitative variations in cell abundance, morphological and physiological conditions were assessed on short-term incubations in semi-continuous cultures, single culture (PE, PC) and co-culture (PE+PC), with and without predators, by flow cytometry, and PhytoPAM. Under grazing pressure, we observed that (i) the abundance of PE single cells decreased over time with a concomitant formation of PE microcolonies; (ii) in PC single cultures, no significant variation in single cells was found and microcolonies did not form; (iii) both PE and PC formed monoclonal microcolonies in co-culture; (iv) PC cells increased the photosynthetic efficiency of the PSII (higher Fv/Fm) in co-culture. In the aftermath of microcolony formation as a predation-induced adaptation, our findings indicated a different response of Synechococcus phylotypes potentially co-existing in natural environment and the importance of their interaction.

The abundance of mixotrophic algae drives the carbon isotope composition of the copepod Boeckella gracilipes in shallow Patagonian lakes

concentrations and the ratio Colorabs440:Chlorophyll a (Chl a). We also determined the isotopic composition of seston and phytoplankton composition. Our results showed a direct relationship between the d 13 C values of B. gracilipes and seston (r 2 ¼ 0.64, P,0.001), and we demonstrated the importance of phytoplankton to B. gracilipes as its d 13 C signal was strongly related to the abundance of the mixotrophic nanoflagellate Chrysochromulina parva (r 2 ¼ 0.86, P , 0.001). We further found an inverse relationship between DOM and the ratio Colorabs440:Chl a and B. gracilipes d 13 C values (r 2 ¼ 0.67 and 0.56, respectively), which also supported the importance of authochthonous carbon for the copepod. We conclude that phytoplankton, particularly mixotrophic algae, rather than allochthonous carbon, supports the copepod B. gracilipes in shallow Patagonian lakes.

Relative importance of phototrophic, heterotrophic, and mixotrophic nanoflagellates in the microbial food web of a river-influenced coastal upwelling area

The contribution of phototrophic nanoflagellates (PNF) as primary producers and het- erotrophic (HNF) and mixotrophic (MNF) nanoflagellates as major grazers of bacterioplankton was assessed during a 3 yr study in a highly productive, river-influenced coastal upwelling area under contrasting seasons (winter/non-upwelling vs. spring/upwelling). Sampling was conducted at 2 stations — around a river plume and at an intensive seasonal upwelling site — with contrasting en- vironmental gradients. The MNF were evaluated functionally for the possession of photo- pigments and experimentally for the ability to take up prey, specifically fluorescently labeled bacteria (FLB). During the short-term experiments, we estimated grazing rates over FLB for 2 size categories of HNF and MNF. Both bacterial production (BP) and nanophytoplankton primary production (PP) were higher in the river plume area. PNF abundance ranged from 6 × 10 9 to 411 × 10 9 cells m �2 , whereas HNF abundance fluctuated between 27 × 10 9 and 267 × 10 9 cells m �2 . In contrast, the abundance of MNF was usually low, with a maximum of ~7 × 10 9 cells m �2 . For MNF, ingestion rates were between 7.3 and 30.7 bacteria per flagellate and hour (bact flag �1 h �1 ), whereas HNF in- gestion ranged from 2 to 7.5 bact flag �1 h �1 . However, since HNF dominated in terms of abundance, they were the dominant grazers on bacterioplankton populations. Estimates of grazing pressure for the microbial food web showed that MNF were capable of removing 1 to 51% BP d �1 , whereas HNF could control BP, eliminating from 24% BP d �1 up to more than 100% BP d �1 . Given the area's relatively high nutrient condition, the elevated MNF biomass in the river plume and the greater bacterivory impact from MNF in winter, it seems that light and, thus, the energy/carbon limitation could be the main trigger for mixotrophy in this river-influenced coastal upwelling area.

UV radiation simultaneously affects phototrophy and phagotrophy in nanoflagellate-dominated phytoplankton from an Andean shallow lake

Mixotrophic nanoflagellates, that combine photosynthesis and phagotrophy, are important members of planktonic food webs in many aquatic environments depending on the balance among the different carbon and energy sources. We carried out field sampling and laboratory experiments with natural nanoflagellate assemblages from an Andean North-Patagonian lake exposing them or not to UVR, and measuring photosynthetic parameters and bacterivory. The effect of different light treatments on the photosynthetic efficiency was studied by the non-invasive, pulse amplitude-modulated (PAM) fluorescence technique, and bacterivory was assessed with fluorescently labeled bacteria (FLB). Mixotrophic nanoflagellates were clearly dominant (up to 90% of total phytoplankton and 88% of total nanoflagellate abundance), and in the experiments labeled bacteria were observed in more than 75% of mixotrophic cells. These results support the idea that these phytoflagellates were never entirely photosynthetic. The high light : phosphorus ratio and the high C : N : P ratio suggest a strong nutrient limitation towards P. Our results show that both functions, photosynthesis and bacteria ingestion, were simultaneously reduced by the same level of UVR. We estimated that UVR exposure of mixotrophic nanoflagellates reduced photosystem II activity between 23% and 31% while ingestion rates were reduced between 23% and 28%. Therefore, our results suggest that the different cell functions could be concurrently impacted by UVR, implying that patterns and rates of C transfer would be substantially altered in the microbial food web.