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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. [ABSTRACT FROM AUTHOR]

In this study, odor characteristics and phytoplankton composition were systematically investigated in two winter periods in a reservoir with fishy odor in north China. Ten potential fishy odor-producing algae were isolated and odorant-producing potentials were evaluated. Olfactometry profile and odorant composition of water samples were analyzed using GC-Olfactometry combined with GC × GC-TOFMS. The results showed that 2,4-heptadienal and hexanal were major fishy odor contributors. The abundance of Uroglena sp., Synura sp. and Peridinium sp. was negatively correlated with total dissolved organic carbon, ammonia nitrogen, and nitrate, illustrating nutrient level might be major drivers for the succession of fishy odor-producing algae. Dinobryon sp. and Uroglena sp. made the greatest contribution to fishy odor, followed by Peridinium sp., Synura sp., and Ochromonas sp. Fishy odor in 2016 winter and the early of 2017 winter was mainly caused by Dinobryon sp., while Uroglena sp. contributes mostly in March in 2017 winter. This study demonstrates the main odorants and algae causing fishy odor in reservoir, which will provide a scientific basis for the management of seasonal fishy odor problems in water source., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2023. Published by Elsevier B.V.)

Waste organics conversion to algal lipid is environmentally friendly and it promotes sustainability. Achieving high‐lipid content in cells is crucial to the economic feasibility of algal lipid production and collection. Here, a phagotrophic microalga Ochromonas danica was grown on waste ketchup and then stimulated to accumulate high‐lipid content using glycerol and acetate. Individually, glycerol and acetate could increase lipid synthesis. Sequential glycerol‐then‐acetate addition was more effective; for an O. danica culture with 0.25 g L−1 cells, the intracellular lipid content was increased to 70–80% (w/w) in 6–12 hours. However, acetate added at >1 g L−1 damaged cells. For high‐density (25 g L−1) fermentations, glycerol and acetate addition strategies were evaluated to overcome this inhibition. Results indicated that glycerol could be added in batches or continuously as long as the amount was sufficient to satisfy the glycerol consumption rate of 6 mg (g cells‐hour)−1 while acetic acid needed to be added continuously, optimal at a rate of 0.27–0.30 g hour−1 or 12–14 mg (g cells‐hour)−1. Intracellular lipid content could reach 70–80% after 2‐day supplementation of glycerol and acetic acid. The methods of glycerol/acetate additions developed here enhance algal lipid production and enable effective conversion of organic wastes to algal lipids as sustainable bioproducts. [ABSTRACT FROM AUTHOR]

Collagenous and elastotic marginal plaques of the hand (CEMPH) is a rare, chronic keratoderma characterized by hyperkeratotic linear plaques located along the radial and ulnar aspects of the hands bilaterally. As an isolated finding, CEMPH occurs secondarily to chronic trauma and photodamage. Herein, CEMPH is described as a manifestation of alkaptonuria (AKU). In addition to keloidal collagen, ochronotic fibers and fragmented, thickened elastic fibers were observed. Additionally, mucin deposition—not previously described in this clinical context—was also identified. Given their overlapping clinicopathologic features, CEMPH due to AKU should be distinguished from the acquired variant as well as acrokeratoelastoidosis. [ABSTRACT FROM AUTHOR]

Cyanobacterial blooms, caused by eutrophication and climate warming, exert severely negative effects on aquatic ecosystem. Some species of protozoans can graze on toxic cyanobacteria and degrade microcystins highly efficiently, which shows a promising way to control the harmful algae. However, in the field, many different species of algae coexist with Microcystis and may affect protozoans eliminating Microcystis . Therefore, in this study, we assessed the impacts of chlorophytes, a type of beneficial algae for zooplankton and common competitors of cyanobacteria, on flagellate Ochromonas eliminating toxin-producing Microcystis at different temperatures. Our results showed that Ochromonas still eliminated Microcystis population and degraded the total microcystins with the addition of chlorophytes, although the time of eliminating Microcystis was prolonged and temperature-dependent. Additionally, in the grazing treatments, chlorophytes populations gradually increased with the depletion of Microcystis , whereas Microcystis dominated in the mixed algal cultures without Ochromonas . The findings indicated that although chlorophytes prolong mixotrophic Ochromonas eliminating Microcystis , the flagellate grazing Microcystis helps chlorophytes dominating in the primary producers, which is significant in improving water quality and reducing aquatic ecosystem risks. [ABSTRACT FROM AUTHOR]

A mixed cyanobacterial-mixotrophic algal population, dominated by the filamentous cyanobacterium Leptolyngbya sp. and the microalga Ochromonas (which contributed to the total photosynthetic population with rates of less than 5%), was studied under non-aseptic conditions for its efficiency to remove organic and inorganic compounds from different types of wastes/wastewaters while simultaneously producing lipids. Second cheese whey, poplar sawdust, and grass hydrolysates were used in lab-scale experiments, in photobioreactors that operated under aerobic conditions with different initial nutrient (C, N and P) concentrations. Nutrient removal rates, biomass productivity, and the maximum oil production rates were determined. The highest lipid production was achieved using the biologically treated dairy effluent (up to 14.8% oil in dry biomass corresponding to 124 mg L−1) which also led to high nutrient removal rates (up to 94%). Lipids synthesized by the microbial consortium contained high percentages of saturated and mono-unsaturated fatty acids (up to 75% in total lipids) for all the substrates tested, which implies that the produced biomass may be harnessed as a source of biodiesel. [ABSTRACT FROM AUTHOR]

Ochromonas spp. strains CCMP1393 and BG-1 are phagotrophic phytoflagellates with different nutritional strategies. Strain CCMP1393 is an obligate phototroph while strain BG-1 readily grows in continuous darkness in the presence of bacterial prey. Growth and gene expression of strain CCMP1393 were investigated under conditions allowing phagotrophic, mixotrophic, or phototrophic nutrition. The availability of light and bacterial prey led to the differential expression of 42% or 45–59% of all genes, respectively. Data from strain CCMP1393 were compared to those from a study conducted previously on strain BG-1, and revealed notable differences in carbon and nitrogen metabolism between the 2 congeners under similar environmental conditions. Strain BG-1 utilized bacterial carbon and amino acids through glycolysis and the tricarboxylic acid cycle, while downregulating light harvesting and carbon fixation in the Calvin cycle when both light and bacteria were available. In contrast, the upregulation of genes related to photosynthesis, light harvesting, chlorophyll synthesis, and carbon fixation in the presence of light and prey for strain CCMP1393 implied that this species is more phototrophic than strain BG-1, and that phagotrophy may have enhanced phototrophy. Cellular chlorophyll a content was also significantly higher in strain CCMP1393 supplied with bacteria compared to those without prey. Our results thus point to very different physiological strategies for mixotrophic nutrition in these closely related chrysophyte species. [ABSTRACT FROM AUTHOR]

Ochromonas triangulata, the type species for a genus with over 125 taxa, was collected for only the second time, again from the type locality. Cell morphology, cell division, palmelloid stage and cyst structure generally agreed with the original description. Molecular phylogenetic analysis based on the 18S rRNA gene revealed 13 clades of Ochromonas-like flagellates as well as the clade represented by our O. triangulata strain and the nearly identical strain RCC-21/AC025. We also conducted a concatenated analysis using the 18S rRNA and the rbcL genes, and we recovered the same 14 clades. One clade, containing strains CCAP 933/27 and CCMP1861, previously named Ochromonas tuberculata, was re-identified as Chrysastrella paradoxa and Chrysastrella breviappendiculata, respectively. One clade included the Poterioochromonas strains but we were unable to convincingly connect species names to the strains because authentic strains were unknown or not examined. Organisms in the clade that included the well-known Ochromonas danica were assigned to Chlorochromonas. The generic name Melkoniania gen. nov. was proposed for one distinct clade of marine flagellates. These changes addressed some of the issues associated with the polyphyletic Ochromonas sensu lato but many problems remained. [ABSTRACT FROM AUTHOR]

Surfactants can affect biological activities and pose serious threats to the aquatic ecosystem if discharged without proper treatment. Rhamnolipid is a biosurfactant with promising agricultural, industrial and biomedical applications. It is important to assess the risks posed by rhamnolipid before it is adopted for large scale use. In this study, toxicity of rhamnolipid was estimated with the effects of motility loss, membrane permeability, and cell lysis using a phagotrophic alga Ochromonas danica. O. danica is a sensitive algal species without a protective cell wall and it represents a group of phagotrophic flagellates ecologically important in the aquatic ecosystem. A common synthetic surfactant, sodium dodecyl sulfate (SDS), was used for comparison. Results suggested that rhamnolipid is significantly less biolytic than SDS and the effects required longer exposure. Lysis of O. danica cells by rhamnolipid was serious only at concentrations over its critical micelle concentration. Motility loss was, however, significantly more sensitive; about 75% loss after 20 min exposure to 50 mg/L rhamnolipid. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the influence of temperature on the ability of the mixotrophic flagellate Ochromonas to eliminate a toxic Microcystis population and degrade microcystins. We exposed Microcystis to cultures with or without Ochromonas YZ1 at 20, 25, and 30 °C for 10 days. Results showed that increased temperature promoted the growth of Ochromonas YZ1 and Microcystis , with the latter achieving high abundance without grazing. With increased temperature, Ochromonas YZ1 clearance rate increased, and Microcystis populations were earlier eliminated. Importantly, Ochromonas YZ1 degraded both intracellular and extracellular microcystins by grazing effects. The reduction ratios of Microcystis abundances and microcystins were both approximately 100% after 6 days at high temperature. In addition, more microcystins were released outside at 20 °C than at the higher temperatures. Overall, this study showed that high temperature favors elimination of toxin-producing Microcystis and degradation of microcystins by mixotrophic Ochromonas . [ABSTRACT FROM AUTHOR]

Combining wastewater treatment with biodiesel production can greatly reduce the cost of environmentally friendly technologies. In this work wastewaters of high environmental importance originating from raisin and wine industries were employed as substrates for a mixed cyanobacterial/algal cultivation system dominated by the filamentous cyanobacterium Leptolyngbya sp. The mixotrophic microalga Ochromonas contributed to less than 5% of the total photosynthetic population and grazed on broken Leptolyngbya trichomes. Satisfactory lipid yields and removal of nutrients and chemical oxygen demand (COD) were observed when raisin or winery wastewaters were used separately as substrates. However, exceptional removal rates were recorded in blends of the above-mentioned wastewaters (i.e., 92.8%, 78.1% and 99% for COD, total nitrogen and phosphate, respectively), while the biomass produced contained around 13% lipids (w/w) on a dry weight basis. The ratio of saturated/mono-unsaturated fatty acids in this lipid reached 85%, making this system suitable for biodiesel production. The proposed system can effectively approach a technology that integrates wastewater treatment with simultaneous biodiesel production. [ABSTRACT FROM AUTHOR]

Background: Ochromonas is a genus of mixotrophic chrysophytes that is found ubiquitously in many aquatic environments. Species in this genus can be important consumers of bacteria but vary in their ability to perform photosynthesis. We studied the effect of light and bacteria on growth and gene expression of a predominantly phagotrophic Ochromonas species. Axenic cultures of Ochromonas sp. were fed with heat-killed bacteria (HKB) and grown in constant light or darkness. RNA was extracted from cultures in the light or in the dark with HKB present (Light + HKB; Dark + HKB), and in the light after HKB were depleted (Light + depleted HKB). Results: There were no significant differences in the growth or bacterial ingestion rates between algae grown in light or dark conditions. The availability of light led to a differential expression of only 8% of genes in the transcriptome. A number of genes associated with photosynthesis, phagotrophy, and tetrapyrrole synthesis was upregulated in the Light + HKB treatment compared to Dark + HKB. Conversely, the comparison between the Light + HKB and Light + depleted HKB treatments revealed that the presence of HKB led to differential expression of 59% of genes, including the majority of genes involved in major carbon and nitrogen metabolic pathways. Genes coding for unidirectional enzymes for the utilization of glucose were upregulated in the presence of HKB, implying increased glycolytic activities during phagotrophy. Algae without HKB upregulated their expression of genes coding for ammonium transporters, implying uptake of inorganic nitrogen from the culture medium when prey were unavailable. Conclusions: Transcriptomic results agreed with previous observations that light had minimal effect on the population growth of Ochromonas sp. However, light led to the upregulation of a number of phototrophy- and phagotrophyrelated genes, while the availability of bacterial prey led to prominent changes in major carbon and nitrogen metabolic pathways. Our study demonstrated the potential of transcriptomic approaches to improve our understanding of the trophic physiologies of complex mixotrophs, and revealed responses in Ochromonas sp. not apparent from traditional culture studies. [ABSTRACT FROM AUTHOR]

Ochromonas sp. (Chrysophyceae) occupies different trophic levels depending on its prevalent nutritional strategy in the field. Ochromonas sp. can utilize inorganic and organic carbon to grow photoautotrophically, mixotrophically, and heterotrophically. To examine how exogenous organic carbon influences population growth and photosynthesis in Ochromonas sp., we cultured Ochromonas sp. in media with different glucose concentrations (0, 50, 100, and 150 mg L) and two treatment modes (single addition of glucose at the beginning and fed-batch mode with repeated addition every 12 h) for 15 days. Results showed that (1) compared to photoautotrophic growth, adding glucose significantly enhanced the growth rate and carrying capacity of Ochromonas sp. in a concentration-dependent manner; (2) photosynthesis also increased with glucose addition during the exponential growth stage; (3) repeated glucose addition caused higher biomass and photosynthetic performances than single addition, and (4) with depletion of glucose and the population reaching a stationary stage, photosynthetic parameters decreased slightly but remained at a relatively stable level, which was similar to that of photoautotrophic growth. We conclude that in comparison with pure photoautotrophy, mixotrophic growth can result in higher population growth rate and final biomass in a shorter time, especially in fed-batch cultures, indicating that this would be a highly efficient way for mass cultivation of the flagellate Ochromonas. [ABSTRACT FROM AUTHOR]

" Spumella "-like flagellates describes similar or even indiscernible colourless non-scaled chrysophytes which are important bacterivores common in different aquatic ecosystems. Recently, phylogenetic analyses revealed a high taxonomic diversity of these flagellates leading to the description of several new genera and species. Our present work on the functional group of pelagic bacterivorous chrysomonads from different water bodies resulted in an extended taxonomic analysis among chrysophytes unveiling yet undescribed genera and species pointing to the high hidden diversity of bacterivores in the pelagial of freshwaters. On the basis of phylogenetic analyses, we describe four new genera Atacamaspumella , Chlorospumella , Pseudapoikia, and Vivaspumella and a new species of the recently described genus Poteriospumella. Beside this, we redescribe the species Ochromonas vasocystis Doflein, 1923 to Poteriospumella vasocystis comb. nov. substantiated on the high sequence similarity with Poteriospumella lacustris Boenigk et Findenig and Poteriospumella maldiviensis nov. sp. [ABSTRACT FROM AUTHOR]

Fucoxanthin (Fx) has attracted great interest due to its remarkable biological activities such as antioxidant and anti-obesity, and its increasing demands in biopharmaceutical and cosmetic fields. However, its commercial production is limited by low yield and high cost. In this study, we isolated and identified a species of golden algae (Ochromonas sp.) capable of engulfing Microcystis aeruginosa (M. aeruginosa) and accumulating Fx. After 72 h mixotrophic cultivation of Ochromonas sp. and M. aeruginosa, the algal culture changed from green to yellow-brown, and the content of Fx and the daily production rate were up to 11.58 mg g -1 , and 1.315 mg L -1 d -1 , respectively. The utilization rate of M. aeruginosa was 527.27 fg cell -1 . This study will not only provide a new thought to produce Fx in an efficient, low-cost, and sustainable way but an innovative method for the control and treatment of harmful cyanobacterial blooms from eutrophic freshwaters as well., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

There is an extensive literature on chrysophyte stomatocysts from diverse habitats and their use as paleolimnetic indicators of past environmental conditions in lakes. The majority of these cysts are unidentified, and more laboratory studies are needed to link stomatocyst morphotypes to vegetative stages. A laboratory procedure that resulted in stomatocyst production in the chrysophyte alga Ochromonas pinguis (Chrysophyceae) was tested to determine if the protocol would also stimulate the production of stomatocysts in six other chrysophytes. Chrysolepidomonas dendrolepidota, Chrysosaccus cf. sphaericus, Dermatochrysis sp., Synura cf. americana, Synura cf. macropora and a second species of Ochromonas all produced stomatocysts within two to three weeks. Morphology of the stomatocyst was described using scanning electron microscopy (SEM). This was the first report of stomatocysts from Chrysosaccus and the first SEM description of O. pinguis cysts. These autecological studies provide a greater linkage between sediment stomatocysts and their biological origin, strengthening the interpretive value of these microfossils and broadening our understanding of chrysophyte biogeography. [ABSTRACT FROM AUTHOR]

In polar regions, melting snow fields can be occupied by striking blooms of chrysophycean algae, which cause yellowish slush during summer. Samples were harvested at King George Island ( South Shetland Islands, Maritime Antarctica) and at Spitsbergen ( Svalbard archipelago, High Arctic). The populations live in an ecological niche, where water-logged snow provides a cold and ephemeral ecosystem, possibly securing the survival of psychrophilic populations through the summer. A physiological adaptation to low temperatures was shown by photosynthesis measurements. The analysis of soluble carbohydrates showed the occurrence of glycerol and sugars, which may play a role in protection against intracellular freezing. Although both populations were made of unicells with Ochromonas-alike morphology, investigation by molecular methods ( 18S rDNA sequencing) revealed unexpectedly a very close relationship to the mountain-river dwelling Hydrurus foetidus ( Villars) Trevisan. However, macroscopic thalli typical for the latter species were never found in snow, but are known from nearby localities, and harvested samples of snow algae exposed to dryness evolved a similar pervading, 'fishy' smell. Moreover, in both habitats tetrahedal zoospores with four elongate spikes were found, similar to what is known from Hydrurus. Our molecular results go along with earlier reports, where chrysophycean sequences of the same taxonomic affiliation were isolated from snow. This points to a distinct group of photoautotrophic, Hydrurus-related chrysophytes, which are characteristic for long-lasting, slowly melting snow packs in certain cold regions of the world. [ABSTRACT FROM AUTHOR]

The metabolic theory of ecology predicts that temperature affects heterotrophic processes more strongly than autotrophic processes. We hypothesized that this differential temperature response may shift mixotrophic organisms towards more heterotrophic nutrition with rising temperature. The hypothesis was tested in experiments with the mixotrophic chrysophyte Ochromonas sp., grown under autotrophic, mixotrophic and heterotrophic conditions. Our results show that (1) grazing rates on bacterial prey increased more strongly with temperature than photosynthetic electron transport rates, (2) heterotrophic growth rates increased exponentially with temperature over the entire range from 13 to 33 °C, while autotrophic growth rates reached a maximum at intermediate temperatures and (3) chlorophyll contents during mixotrophic growth decreased at high temperature. Hence, the contribution of photosynthesis to mixotrophic growth strongly decreased with temperature. These findings support the hypothesis that mixotrophs become more heterotrophic with rising temperature, which alters their functional role in food webs and the carbon cycle. [ABSTRACT FROM AUTHOR]

We investigated the selective predation of mixotrophic and heterotrophic flagellates (MF and HF) on different heterotrophic prokaryote phylotypes (HPP; Bacteria + Archaea) living in natural assemblages from oligotrophic Antarctic lakes. In situ prey preference was analyzed for the first time on different mixotrophic taxa (Pseudopedinella sp., Ochromonas-like cells, Chrysophyceae >5 µm). The relative abundances of seven different HPP hybridized by CARD-FISH (catalyzed reporter deposition-fluorescent in situ hybridization) in natural community were compared with the proportions of hybridized cells inside digestive vacuoles. Our results showed some general trends to selectivity over some HPP. Alphaproteobacteria and Betaproteobacteria were the most abundant groups, and strikingly, a negative selection trend was detected in most samples by all bacterivorous protists. In contrast, for Actinobacteria a positive selection trend was observed in most samples, whereas Bacteroidetes seemed to be randomly preyed upon. Interestingly, similar prey preferences were observed in all bacterivorous flagellates. Our results suggest that phylogenetic affiliation determines part of the process of prey selection by protists in these lakes. Nevertheless, other features, such as cell size, morphology and the presence of the S-layer, might also significantly contribute to prey selectivity on the HPP. [ABSTRACT FROM AUTHOR]

1. Toxic compounds produced by many phytoplankton taxa are known to have negative effects on competitors (allelopathy), anti-predatory effects on grazers (mortality or impaired reproduction) or both. Although mixotrophs of the genus Ochromonas are known to be toxic to zooplankton, it has often been assumed in studies of plankton community processes that all flagellates in the size range of this taxon are edible to typical zooplankton grazers (i.e. cells ≤30 μm for Daphnia, ≤6 μm for rotifers). 2. We explored the toxicity of a species of Ochromonas to other planktonic taxa, including its competitors (two species of phytoplankton and protists) and consumers (two species of zooplankton). To test if mode of nutrition by this mixotroph influences its toxicity to other taxa, we exposed each test species to Ochromonas cultured in chemostats under four different nutritional regimes: osmotrophy (labile dissolved organic carbon) and phagotrophy (bacterial prey) in both light and dark conditions (i.e. with or without photosynthesis). 3. Filtrate from osmotrophically fed Ochromonas had a significant negative effect on the population growth rate of two obligate phototrophic phytoplankton, Cryptomonasozolini and Chlamydomonas reinhardtii. The protists Tetrahymena tetrahymena and Paramecium aurelia were also negatively affected by Ochromonas filtrate. Ochromonas cells were toxic to both the rotifer Brachionus calicyflorus and the cladoceran Daphnia pulicaria, with the toxic effects significantly more severe when fed at high cell densities (75 000 cells mL−1) than at low densities (7500 cells mL−1). Ochromonas cultured osmotrophically in the light was more toxic to the Daphnia than cells cultured under other conditions. In contrast, Ochromonas from all nutritional conditions was equally highly toxic to Brachionus. 4. Our findings support the view that Ochromonas can be toxic to other components of the food web with which it interacts. It is especially toxic to zooplankton that directly consume it, although the effect depends upon Ochromonas cell density and whether or not a good food source is simultaneously present. Our results call into question the common practice of pooling flagellates into a single 'functional group' included in an 'edible phytoplankton' category of cells <30 μm in diameter. [ABSTRACT FROM AUTHOR]

F-actin organization during the cell cycle was investigated in two stramenopile microalgae, Ochromonas danica (Chrysophyceae; UTEX LB1298) and Heterosigma akashiwo (Raphidophyceae; NIES-6) using FITC-phalloidin. In the interphase cell of O. danica, F-actin bundles were localized forming a network structure in the cortical region, which converged from the anterior region to the posterior, whereas in the interphase cell of H. akashiwo, F-actin bundles were observed forming a network structure in the cortical region without any polarity. In both O. danica and H. akashiwo, at the initial stage of mitosis the cortical F-actin disappeared, and then during cytokinesis assembly of an actin-based ring-like structure occurred in the cell cortex in the plane of cytokinesis. The ring-like structure initiated from aster-like structures was composed of F-actin in both O. danica and H. akashiwo. Different from animal cells, later stages of cytokinesis of O. danica seemed to be promoted by microtubules, although the early stages of cytokinesis progressed with a constriction of the ring-like structure, whereas cytokinesis of H. akashiwo was apparently completed by constriction of the cell mediated by the F-actin ring, as in animal cells. [ABSTRACT FROM AUTHOR]

Induced colony formation under grazing pressure has been reported in our previous results. However, the colonies induced in these studies comprised only tens of cells which are far smaller than the naturally occurring colonies. In this work, unicellular Microcystis aeruginosa Kützing were co-cultivated with flagellate Ochromonas sp. for 50 d to investigate colony formation in M. aeruginosa under continuous grazing pressure. Results revealed that colonial M. aeruginosa formed on the 10th d under the grazing pressure of flagellate. These algal colonies resulted from the daughter cells of freshly dividing cells that failed to separate during the reproductive process. The diameters and cell numbers of the colonies increased slowly with time. Under continuous grazing pressure by Ochromonas sp. for 50 d, the diameter of some colonies reached over 180 µm. Analysis showed that the extra cellular polysaccharide (EPS) content and relative gas vesicle (RGV) of each cell increased significantly after colony formation. However, there was no significant difference on the monosaccharide composition between unicellular and colonial M. aeruginosa. The loose aggregation of cells in the floating colonies suggests that a correlation probably exists between cell compactness and colony buoyancy. [ABSTRACT FROM AUTHOR]

Strains of the green alga Chlamydomonas acidophila and two chrysomonads, Ochromonas spp., isolated from each of two similar acid mining lakes (AMLs) with extremely low pH (∼2.6) were investigated to consider a possible synergistic stress effect of low pH and unfavourable temperature. We measured flagellate growth rates over a combination of four pH (2.5, 3.5, 5.0 and 7.0) and three temperatures (10, 17.5 and 25°C) in the laboratory. Our hypothesis was that, under highly acidic conditions (pH <3), an obligate acidophil species (C. acidophila) would be less sensitive to the combined stress of pH and temperature than acidotolerant species (Ochromonas spp.). We expected that the difference of the fundamental vs. realized pH niche would be greater in the latter. Another chrysomonad, Poterioochromonas malhamensis strain DS, served as a reference for a closely related neutrophil species. Surprisingly, C. acidophila did not survive temperatures >27°C. The lowest temperature tested reduced growth rates of all three chrysomonad strains significantly. Since all chrysomonads were tolerant to high temperature, growth rate of one Ochromonas spp. strain was measured exemplarily at 35°C. Only at this high temperature was the realized pH niche significantly narrowed. We also recorded significant intraspecific differences within the C. acidophila strains from the two AML, illustrating that the niche width of a species is broader than that of individual clones. [ABSTRACT FROM AUTHOR]

Abstract: We investigated the effect of pH on the competition of two closely related chrysomonad species, Poterioochromonas malhamensis originating from circumneutral Lake Constance, and Ochromonas sp. isolated from a highly acidic mining lake in Austria (pH ∼2.6). We performed pairwise growth experiments between these two species at four different pH ranging from 2.5 to 7.0. Heterotrophic bacteria served as food for both flagellates. Results were compared to growth rates measured earlier in single species experiments over the same pH range. We tested the hypothesis that the acidotolerant species benefits from competitive release under conditions of acid stress. The neutrophilic strain numerically dominated over the acidotolerant strain at pH 7.0, but was the inferior competitor at pH 2.5. At pH 3.5 and 5.0 both strains coexisted. Surprisingly, P. malhamensis prevailed over Ochromonas sp. under moderately acidic conditions, i.e. at the pH where growth rates of the latter peaked when grown alone. Since bacterial food was not limiting, resource competition is improbable. It appears more likely that P. malhamensis ingested cells of its slightly smaller competitor. Adverse effects mediated via allelopathy, either directly on the competing flagellate or indirectly by affecting its bacterial food, might also have affected the outcome of competition. [Copyright &y& Elsevier]

Snow algae inhabit unique environments such as alpine and high latitudes, and can grow and bloom with visualizing on snow or glacier during spring-summer. The chrysophytes Ochromonas smithii and Ochromonas itoi are dominant in yellowcolored snow patches in mountainous heavy snow areas from late May to early June. It is considered to be effective utilizing the xanthophyll cycle and holding sunscreen pigments as protective system for snow algae blooming in the vulnerable environment such as low temperature and nutrients, and strong light, however the study on the photoprotection of chrysophytes snow algae has not been shown. To dissolve how the chrysophytes snow algae can grow and bloom under such an extreme environment, we studied with the object of light which is one point of significance to this problem. We collected the yellow snows and measured photosynthetically active radiation at Mt. Gassan in May 2008 when the bloom occurred, then tried to establish unialgal cultures of O. smithii and O. itoi, and examined their photosynthetic properties by a PAM chlorophyll fluorometer and analyzed the pigment compositions before and after illumination with high-light intensities to investigate the working xanthophyll cycle. This experimental study using unialgal cultures revealed that both O. smithii and O. itoi utilize only the efficient violaxanthin cycle for photoprotection as a dissipation system of surplus energy under prolonged high-light stress, although they possess chlorophyll c with diadinoxanthin. [ABSTRACT FROM AUTHOR]

Background: The need for new therapies for cholangiocarcinoma is highlighted by their poor prognosis and refractoriness to chemotherapy. Increased production of Interleukin-6 promotes cholangiocarcinoma growth and contributes to chemoresistance by activating cell survival mechanisms. We sought to identify biologically active compounds capable of ameliorating the phenotypic effects of IL-6 expression and to explore their potential therapeutic use for cholangiocarcinoma. Methodology: A genomic signature associated with Interleukin-6 expression in Mz-ChA-1 human malignant cholangiocytes was derived. Computational bioinformatics analysis was performed to identify compounds that induced inverse gene changes to the signature. The effect of these compounds on cholangiocarcinoma growth was then experimentally verified in vitro and in vivo. Interactions with other therapeutic agents were evaluated using median effects analysis. Principal Findings: A group of structurally related compounds, nitrendipine, nifedipine and felodipine was identified. All three compounds were cytotoxic to Mz-ChA-1 cells with an IC50 for felodipine of 26 μM, nitrendipine, 44 μM and nifedipine, 15 μM. Similar results were observed in KMCH-1, CC-LP-1 and TFK-1 cholangiocarcinoma cell lines. At a fractional effect of 0.5, all three agents were synergistic with either camptothecin or gemcitabine in Mz-ChA-1 cells in vitro. Co-administration of felodipine and gemcitabine decreased the growth of Mz-ChA-1 cell xenografts in nude athymic mice. Conclusions: Computational bioinformatics analysis of phenotype-based genomic expression can be used to identify therapeutic agents. Using this drug discovery approach based on targeting a defined tumor associated phenotype, we identified compounds with the potential for therapeutic use in cholangiocarcinoma. [ABSTRACT FROM AUTHOR]

The behavior and toxicity of silver engineered nanoparticles (Ag-ENs) to the mixotrophic freshwater alga Ochromonas danica were examined in the present study to determine whether any other mechanisms are involved in their algal toxicity besides Ag+ liberation outside the cells. Despite their good dispersability, the Ag-ENs were found to continuously aggregate and dissolve rapidly. When the initial nanoparticle concentration was lower than 10 μM, the total dissolved Ag+ concentration ([Ag+]T) in the suspending media reached its maximum after 1 d and then decreased suggesting that Ag+ release might be limited by the nanoparticle surface area under these conditions. Furthermore, Ag-EN dissolution extent remarkably increased in the presence of glutathione. In the Ag-EN toxicity experiment, glutathione was also used to eliminate the indirect effects of Ag+ that was released. However, remarkable toxicity was still observed although the free Ag+ concentration in the media was orders of magnitude lower than the non-observed effect concentration of Ag+ itself. Such inhibitive effects were mitigated when more glutathione was added, but could never be completely eliminated. Most importantly, we demonstrate, for the first time, that Ag-ENs can be taken in and accumulated inside the algal cells, where they exerted their toxic effects. Therefore, nanoparticle internalization may be an alternative pathway through which algal growth can be influenced. [ABSTRACT FROM AUTHOR]

Ecological stoichiometry focuses on the balance between multiple nutrient elements in resources and in consumers of those resources. The major consumers of bacteria in aquatic food webs are heterotrophic and mixotrophic nanoflagellates. Despite the importance of this consumer-resource interaction to understanding nutrient dynamics in the aquatic food web, few data are available addressing the element stoichiometry of flagellate consumers. Ochromonas danica, a mixotrophic bacterivore, was used as a model organism to study the relationships among temperature, growth rate and element stoichiometry. Ochromonas danica was grown in chemostats at dilution rates ranging between 0.03 and 0.10 h−1 and temperatures ranging between 15 and 28 °C. Cells accumulated elements as interactive functions of temperature and growth rate, with the highest element concentrations corresponding to cells grown at a low temperature and high growth rates. The highest concentrations of elements were associated with small cells. Temperature and growth rate affected the element stoichiometry (as C : N, C : P and N : P) of O. danica in a complex manner, but the growth rate had a greater effect on ratios than did temperature. [ABSTRACT FROM AUTHOR]

Abstract: Acidic mining lakes (pH <3) are specific habitats exhibiting particular chemical and biological characteristics. The species richness is low and mixotrophy and omnivory are common features of the plankton food web in such lakes. The plankton community structure of mining lakes of different morphometry and mixing type but similar chemical characteristics (Lake 130, Germany and Lake Langau, Austria) was investigated. The focus was laid on the species composition, the trophic relationship between the phago-mixotrophic flagellate Ochromonas sp. and bacteria and the formation of a deep chlorophyll maximum along a vertical pH-gradient. The shallow wind-exposed Lake 130 exhibited a higher species richness than Lake Langau. This increase in species richness was made up mainly by mero-planktic species, suggesting a strong benthic/littoral – pelagic coupling. Based on the field data from both lakes, a nonlinear, negative relation between bacteria and Ochromonas biomass was found, suggesting that at an Ochromonas biomass below 50μgCL−1, the grazing pressure on bacteria is low and with increasing Ochromonas biomass bacteria decline. Furthermore, in Lake Langau, a prominent deep chlorophyll maximum was found with chlorophyll concentrations ca. 50 times higher than in the epilimnion which was build up by the euglenophyte Lepocinclis sp. We conclude that lake morphometry, and specific abiotic characteristics such as mixing behaviour influence the community structure in these mining lakes. [Copyright &y& Elsevier]

Two-dimensional (2-D) protein analysis of the mastigoneme fraction of the chromophyte alga Ochromonas danica E. G. Pringsh. showed the presence of several component proteins of the tubular mastigoneme. Adding to the reported gene Ocm1, three new genes ( Ocm2, Ocm3, and Ocm4) belonging to the Ocm gene family were isolated using degenerate primers designed from predicted Ocm1 amino acid sequences. The predicted polypeptides encoded by Ocm2, Ocm3, and Ocm4 were smaller in size than Ocm1. However, they shared four highly conserved, cysteine-rich, epithelial growth factor (EGF)-like motifs, potentially involved in protein–protein interaction. In addition, Ocm2, Ocm3, and Ocm4 showed homology to the SIG protein family in the centric diatom Thalassiosira weissflogii (Grunow) Fryxell et Hasle, which is up-regulated during early stages of sexual reproduction. Immunofluorescence analysis with a polyclonal antibody against the partial amino acid sequences of Ocm2, Ocm3, and Ocm4 showed that Ocm2 and Ocm3 were located in the basal segment region of mastigonemes attached on the surface of the anterior flagellum, and that Ocm4 was located within the tubular shaft portion similar to Ocm1. [ABSTRACT FROM AUTHOR]

1. In order to test the effect of Ochromonas sp. , a mixotrophic chrysophyte, on cyanobacteria, grazing experiments were performed under controlled conditions. We studied grazing on three Microcystis aeruginosa strains, varying in toxicity and morphology, as well as on one filamentous cyanobacterium, Pseudanabaena sp. Furthermore, we analysed the co-occurrence of Ochromonas and Microcystis in natural systems in relation to various environmental parameters (TP, TN, DOC, temperature, pH), using data from 460 Norwegian lakes. 2. Ochromonas was able to feed on all four cyanobacterial strains tested, and grew quickly on all of them. The chrysophyte caused net growth reductions in all three Microcystis strains (the very toxic single-celled strain PCC 7806; the less toxic colony-forming Bear AC and the less toxic single-celled Spring CJ). The effect of Ochromonas was strongest on the Spring CJ strain. Although the effect of Ochromonas grazing on the growth of Pseudanabaena was relatively smaller, it also reduced the net growth of this cyanobacterium significantly. 3. After 4 days of incubation with Ochromonas the total amount of cyanotoxins in the three Microcystis strains was reduced by 91.1–98.7% compared with the controls. 4. Ochromonas occurred in similar densities across all 460 Norwegian lakes. Microcystis occurred only at higher TN, TP, temperature and pH values, although its density was often several orders of magnitude higher than that of Ochromonas. Ochromonas co-occurred in 94% of the samples in which Microcystis was present. 5. From our study it is not clear whether Ochromonas could control Microcystis blooms in natural lakes. However, our study does demonstrate that Ochromonas usually occurs in lakes with Microcystis, and our small scale experiments show that Ochromonas can strongly reduce the biomass of Microcystis and its toxin content. [ABSTRACT FROM AUTHOR]

We investigated grazing and specific growth rates of the mixotrophic flagellate (MxFI) Ochromonas sp. when fed toxic Microcystis aeruginosa (NIES-88, NIES-298, and NIES-44 strains) and a non-toxic strain (NIES-101). The maximum growth rates of Ochromonas sp. were 1.28 d-1 for M. aeruginosa NIES-44, 1.24 d-1 for NIES-88, 2.05 d-1 for NIES-298, and 1.53 d-1 for NIES-101. Also, the maximum ingestion rates on each Microcystis strain by Ochromonas sp. were 4.32 prey MxFl-1 d-1 for M. aeruginosa NIES-44, 5.71 prey MxFl-1 d-1 for NIES-88, and 2.12 prey MxFI-1 d-1 for NIES-101. Interestingly, although the growth rate of Ochromonas sp. with the NIES-298 strain was higher than it was for any of the other strains, its ingestion rate for NIES-298 was low and similar to that with NIES-101. There are two possible explanations for this difference. First, the colony formation by the NIES-298 and NIES-101 strains may have interfered with ingestion. Second, the medium nutrients available to Ochromonas sp. were sufficient to support its population growth despite low grazing on prey, since this flagellate gains its nutrients through a combination of autotrophy and phagotrophy. The flagellate Ochromonas sp. showed active growth regardless of toxic and non-toxic prey types. [ABSTRACT FROM AUTHOR]

Abstract: The composition and distribution of the main planktonic halophilic micro-organisms (heterotrophic and autotrophic picoplankton, nanoplankton, phytoplankton, ciliates) and metazooplankton were investigated in six ponds of increasing salinity in the solar salt works of Sfax, Tunisia, from January to December 2003. Marked changes in the composition and biomass of the communities were found along the salinity gradient, especially at salinities of 150 and 350. Autotrophic picoplankton, nanoplankton, diatoms, dinoflagellates and ciliates characterized the less salted ponds. Planktonic biomass was the highest at intermediate salinity as a consequence of a bloom of Ochromonas. Species richness of phytoplankton, ciliates and zooplankton greatly decrease above a salinity of 150 and typical halophiles (Dunaliella salina, cyanobacteria, Fabrea salina and Artemia salina) were found between 150 and 350 salinity. In this environment, F. salina appeared more adapted than the brine shrimp to survive during phytoplankton blooms. The halophilic plankton was however almost entirely composed of heterotrophic prokaryotes in the crystallizers. We thus observed a progressive disappearance of the autotrophic planktonic communities along the salinity gradient. Multivariate analysis of the communities provides evidence that ponds represent discrete aquatic ecosystems within this salt works. [Copyright &y& Elsevier]

Removal of harmful cyanobacteria is an extremely urgent task in global lake management and protection. Conventional measures are insufficient for simultaneously removing cyanobacteria and hazardous cyanotoxin, efficient and environmental-friendly measures are therefore particularly needed. Herbivorous protozoa have great potentials in controlling algae, however, large-sized colonial Microcystis is inedible for protozoa, which is a central problem to be solved. Therefore, in present study, a measure of protozoa grazing assisted by ultrasound was investigated in laboratory scale for eliminating harmful colonial Microcystis. The results showed that with ultrasound power and time increasing, the proportion of unicellular Microcystis increased significantly. With Ochromonas addition, approximately 80% of colonial Microcystis and microcystin was removed on day 4 under ultrasound power of 100 W for 15 min, while Ochromonas only reduced Microcystis by less than 20% without assistance of ultrasound. Moreover, when directly exposed to low-intensity ultrasound, Ochromonas showed strong resistance to ultrasound and were not inhibited in grazing Microcystis. Overall, ultrasound increases edible food for protozoa via collapsing Microcystis colonies and assists Ochromonas to remove Microcystis , thus intermittently collapsing colonial Microcystis using low-intensity ultrasound can significantly improve the removal efficiency of Microcystis by protozoa grazing, which provided a new insight in controlling harmful colonial Microcystis. [Display omitted] • Low-intensity ultrasound disaggregates Microcystis colonies but avoids cell lysis. • Increasing ultrasonic power and time increase edible Microcystis for Ochromonas. • Colonial Microcystis and MCs can be removed by Ochromonas assisted by ultrasound. • Ochromonas can resist low-intensity ultrasound and its ingestion are not affected. • Ultrasound condition is the key factor affecting Microcystis removal efficiency. [ABSTRACT FROM AUTHOR]

We document a remarkable morphological transformation, attributable to grazing by nanoflagellate Ochromonas sp. DS , of a phycocyanin-rich freshwater Cyanobium sp. (10-NR 98.2% similar 16S rRNA gene sequence to the type species Cyanobium gracile) . The single cells aggregated into microcolonies (average size 40 μm) in the presence of the protist. Colonies were characterized by hundreds of tubes (spinae), 100 nm to 1 μm long and 63 ± 6 nm wide, on the surfaces of the Cyanobium cells co-cultured with Ochromonas. Spinae production, previously unknown for the freshwater Cyanobium species, suggests that picocyanobacterial life strategies are more flexible than previously thought. [ABSTRACT FROM AUTHOR]

The phylogenetic group stramenopiles refers to the systematic groups that possess tripartite tubular hairs (stramenopiles) on their flagella. There have been a number of studies describing the fine structure of these mastigonemes and a few studies isolating the component proteins; however, these proteins and their gene sequences have not yet been identified. In the present study, we identified a mastigoneme protein (Ocm1) of the chrysophycean alga Ochromonas danica Pringsh. (UTEX LB1298). Its corresponding gene, Ocm1, was identified by using degenerate primers that correspond to the partial amino acid sequences of a protein (85 kDa) obtained from a mastigoneme-rich fraction of isolated flagella. The polypeptide encoded by Ocm1 has four cysteine-rich, epithelial growth factor (EGF)–like motifs, potentially involved in protein–protein interactions. It lacks obvious hydrophobic regions characteristic of transmembrane domains, suggesting that this polypeptide is not likely a protein for anchoring the mastigoneme. In addition, a polyclonal antibody against Ocm1 labeled the area where the tubular shafts of the mastigonemes are located, but not the basal portion or the terminal filaments. [ABSTRACT FROM AUTHOR]

Predation and competition are two main factors that determine the size and composition of aquatic bacterial populations. Using a simplified bacterial community, composed of three strains characterized by different responses to predation, a short-term laboratory experiment was performed to evaluate adaptations and relative success in communities with experimentally controlled levels of predation and nutrient availability. A strain with a short generation time ( Pseudomonas putida), one with high plasticity in cell morphology ( Flectobacillus sp. GC5), and one that develops microcolonies ( Pseudomonas sp. CM10), were selected. The voracious flagellate Ochromonas sp. was chosen as a predator. To describe adaptations against grazing and starvation, abundance, biomass and relative heterogeneity of bacteria were measured. On the whole, the strains in the predation-free cultures exhibited unicellular growth, and P. putida represented the largest group. The presence of Ochromonas strongly reduced bacterial abundance, but not always the total biomass. The activity of grazers changed the morphological composition of the bacterial communities. Under grazing pressure the relative composition of the community depended on the substrate availability. In the presence of predators, P. putida abundance declined in both high and low nutrient treatments, and Pseudomonas CM10 developed colonies. Flectobacillus was only numerically codominant in the nutrient-rich environments. [ABSTRACT FROM AUTHOR]

Phagotrophy and competitive ability of the mixotrophic Ochromonas minima were investigated in a three-factorial experiment where light intensity (low: 1.0μmolm−2 s−1 and high: 60μmolm−2 s−1 PPFD), nutrient concentration (ambient: 7.0μmolNl−1, 0.11μmolPl−1 and enriched: 88μmolNl−1, 6.3μmolPl−1) and DOC supply (without and with enrichment, 250μmolCl−1) were manipulated. Ochromonas minima and bacterial abundance were monitored for 12 days. We found significant and interacting effects of light and nutrients on Ochromonas minima growth rate and abundance. At high light intensity, nutrient enrichment resulted in increased growth rates and population sizes. In contrast, reduced growth rates and population sizes were observed for nutrient enrichment when light intensity was low. Although, Ochromonas minima was able to ingest bacteria under both high and low light conditions, it grew only when light intensity was high. At high light intensity, Ochromonas minima grew exponentially under nutrient conditions that would have been limiting for photoautotrophic microalgae. In non-enriched low light treatments, Ochromonas minima populations survived, probably by using background DOC as an energy source, indicating that this ability can be of relevance for natural systems even when DOC concentrations are relatively low. When competing with photoautotrophic microalgae, the ability to grow under severe nutrient limitation and to survive under light limitation should be advantageous for Ochromonas minima. [Copyright &y& Elsevier]

In the experiment we investigated the effect of grazing by different sorts of zooplankton on the induction of defensive morphology in the cyanobacterium Microcystis aeruginosa. The results showed that protozoan flagellate Ochromonas sp. grazing could induce colony formation in M. aeruginosa, whereas M. aeruginosa populations in the control and the grazing treatments of copepod Eudiaptomus graciloides, cladoceran Daphnia magna, and rotifer Brachionus calyciflorus were still strongly dominated by unicells and paired cells and no colony forma occurred. In the protozoan grazing treatment, the proportion of unicells reduced from 83.2% to 15.7%, while the proportion of cells in colonial form increased from 0% to 68.7% of the population at the end of the experiment. The occurrence of a majority of colonial M. aeruginosa being in the treatment with flagellates, indicated that flagellate grazing on solitary cells could induce colony formation in M. aeruginosa. The colonies could effectively deter flagellate from further grazing and thus increase the survival of M. aeruginosa. The colony formation in M. aeruginosa may be considered as an inducible defense against flagellate grazing under the conditions that toxin cannot deter flagellate from grazing effectively. [ABSTRACT FROM AUTHOR]

Mixotrophic organisms combine light, mineral nutrients, and prey as supplementary resources. Based on theoretical assumptions and field observations, we tested experimentally the hypothesis that mixotrophs may invade established plankton communities depending on the trophic status of the system, and investigated possible effects on food web structure, species diversity, and nutrient dynamics. To test our hypothesis, we inoculated the mixotrophic nanoflagellate Ochromonas tuberculata into established planktonic food webs, consisting of specialist phototrophs, specialist phagotrophs, and bacteria at different supplies of soluble inorganic nutrients and dissolved organic carbon. Oligotrophic systems facilitated the invasion of O. tuberculata in two different ways. First, the combination of photosynthesis and phagotrophy gave mixotrophs a competitive advantage over specialist phototrophs and specialist phagotrophs. Second, low nutrient supplies supported the growth of small plankton organisms that fell into the food size spectrum of mixotrophs. Conversely, high nutrient supplies prevented O. tuberculata from successfully invading the food webs. Two important conclusions were derived from our experiments. First, in contrast to a paradigm of ecology, specialization may not necessarily be the most successful strategy for survival under stable conditions. Indeed, the use of several resources with lower efficiency can be an equally, or even more, successful strategy in nature. Second, when limiting nutrients promote the growth of bacterio- and picophytoplankton, invading mixotrophs may have a habitat-ameliorating effect for higher trophic levels, gauged in terms of food quantity and quality. Using given resources more efficiently, O. tuberculata generated higher biomasses and expressed an increased nutritional value for potential planktivores, due to decreased cellular carbon to phosphorus (C:P) ratios compared to specialized plankton taxa. Our findings may help to explain why energy transfer efficiency between phytoplankton and higher trophic levels is generally higher in oligotrophic systems than in nutrient rich environments. [ABSTRACT FROM AUTHOR]

We reared mixotrophic (Ochromonas tuberculata and Cryptomonas sp.) and photoautotrophic specialist algae (Scenedesmus obliquus) at different light:phosphorus supplies and compared their effects as food for zooplankton (Daphnia magna). According to the light:nutrient hypothesis (LNH), biomass and nutrient stoichiometry of phototrophic specialists depend strongly on light:phosphorus supplies. If this is true, herbivore growth and fecundity should be limited by food quantity at low light intensities and by stoichiometric food quality at high light intensities. In turn, phosphorus fertilization should cause a transition from limitation by food quality to limitation by food quantity. In contrast to the LNH, biomass and nutrient stoichiometry of mixotrophs were almost unaffected by alterations in the supply of light and dissolved nutrients. Bacterial counts indicate that mixotrophs compensated for light or phosphorus deficiency by heterotrophic nutrition. Compared to phototrophic specialists, a diet of Cryptomonas sp. therefore enabled a similar or higher and more stable secondary production at most light: nutrient supplies. O. tuberculata, however, appeared to be toxic. Our results indicate that mixotrophs might have a balancing effect on variations in transfer efficiency caused by perturbations to light and nutrient supplies. [ABSTRACT FROM AUTHOR]

Centrin, the EF-hand Ca2+-binding protein is localized at the basal apparatus of flagella and in centrioles in many eukaryotic cells. In the present study, centrin genes of the heterokont algae have been clarified for the first time. We isolated and analyzed cDNA and genomic DNA of centrin genes from the crysophycean algaOchromonas danicaPrings (UTEX LB1298) and the brown algaScytosiphon lomentaria(Lyngbye) Link. The centrin gene ofOchromonascontained an open reading frame of 163 amino acids. The deduced protein, named Odcen, exhibited 85%, 78% and 59% homology toChlamydomonas, human andArabidopsiscentrin, respectively. The centrin genes ofScytosiphoncontained an open reading frame of 164 amino acids. The deduced protein, named Slcen, exhibited 84%, 77% and 59% homology toChlamydomonas, human andArabidopsiscentrin, respectively. Both Odcen and Slcen possessed N-terminal extensions before the conserved amino acid among various centrins, four EF-hand domains and an aromatic amino acid at the C-terminus. Southern blot hybridization suggested that the centrin gene occurs as a single copy gene in bothOchromonasandScytosiphongenomes. Comparison of the sequence of the cDNA and the genomic DNA revealed that the Odcen gene was split into three fragments by introns and Slcen gene consisted of five fragments. The junctions of all introns of both genes conformed to the GT-AG rule. The introns of Slcen gene were considerably long and, as a result, the Slcen gene was approximately seven times longer than Odcen gene. [ABSTRACT FROM AUTHOR]

Current models suggest that (i) flamentous bacteria are protected against predation by nanoflagellates, (ii) prey size is positively correlated with prey-predator contact probability, and (iii) contact probability is mainly responsible for size-selective predation by interception-feeding flagellates. We used five strains of filamentous bacteria and one bacterivorous nanoflagellate, Ochromonas sp. strain DS, to test these assumptions. The five strains, including one spirochete and four Betaproteobacteria strains, were isolated by the filtration-acclimatization method. All five strains possess flexible cells, but they differ in average cell length, which ranged from 4.5 to 13.7 µm. High-resolution video microscopy was used to measure contact, capture, and ingestion rates, as well as selectivity of the flagellate feeding. Growth and feeding experiments with satiating and nonsatiating food conditions, as well as experiments including alternative well-edible prey, were performed. In contrast to predictions by current models, the flagellate successfully consumed all the tested filamentous strains. The ingestion rate was negatively correlated with bacterial length. On the other hand, the lengths of the filamentous bacteria were not positively correlated to the contact rate and capture rate but were negatively correlated to ingestion efficiency. In experiments including alternative nonfilamentous prey, the flagellates showed negative selection for filamentous bacteria, which was independent of food concentration and is interpreted as a passive selection. Our observations indicate that (i) size alone is not sufficient to define a refuge for filamentous bacteria from nanoflagellate predation and (ii) for the investigated filamentous bacteria, prey-predator contact probability could be more influenced by factors other than the prey size. [ABSTRACT FROM AUTHOR]

We examined the impact of nutrient conditions (carbon and phosphorus limitation) and grazing by protozoans on the phenotypic community structure of freshwater bacteria in continuous culture systems. Lakewater bacteria were grown on mineral medium, which was supplemented with glucose and amino acids and adjusted by different phosphorus concentrations to achieve either carbon or phosphorus limitation. Each nutrient treatment was inoculated with the same bacterial community and consisted of a nongrazing and a grazing treatment, to which the heterotrophic nanoflagellates Spumella sp. and Ochromonas sp. were added. We found that nutrient conditions alone resulted in differences in the phenotypic structure of the bacterial community: small and motile bacteria dominated under C limitation while large, elongated, and capsulated bacteria were characteristic for P limitation. The genotypic community composition as measured by T-RFLP (terminal restriction fragment length polymorphism) was not severely influenced by the two nutrient treatments. In the presence of flagellate predators, grazing-resistant bacteria developed under both nutrient conditions, but with different survival mechanisms: Highly motile bacteria prevailed under C limitation, whereas the P-limited grazing treatment was dominated by filamentous forms. T-RFLP analysis revealed only moderate changes in bacterial community composition due to grazing, which were most pronounced under P limitation. Analysis by video microscopy revealed that high swimming speed is an efficient nonmorphological survival mechanism for bacteria to reduce the capture success of the flagellate predator. The rejection of optimal-sized, nonmotile bacteria under P limitation suggests the importance of other nonmorphological, surface-located cell properties. Our results illustrate that the realized mechanisms of grazing resistance are linked to the actual limitation conditions, and that the combined effects of nutrient limitation and grazing are major determinants of bacterial community structure. [ABSTRACT FROM AUTHOR]

Axenic growth of a mixotrophic alga, Ochromonas sp., was compared in several inorganic and organic media, and in the presence of live bacteria under nutrient-replete and low-nutrient conditions. Axenic growth in the light was negligible in inorganic media with or without the addition of glucose. Addition of vitamins increased growth rate, but average cell size declined, resulting in no net increase in biomass. Supplementing axenic cultures with a more complex organic substrate resulted in moderate growth and higher maximal abundance (and biomass) than in the inorganic media with added vitamins. The absence of light did not greatly affect population growth rate in the presence of complex dissolved organic compounds, although cell size was significantly greater in the light than in the dark. The highest growth rates for the alga (up to 2.6 d?1) were measured in treatments containing live bacteria. Increases in cell number of Ochromonas sp. in the presence of bacterial prey were similar in the light and dark, although chloroplast and cell sizes differed. Bacterial abundance was reduced and dissolved phosphorus and ammonia were rapidly released in bacterized cultures in the light and dark, indicating high rates of bacterial ingestion and suggesting an inability of the alga to store or utilize N and P in excess of the quantities required for heterotrophic growth. Low-nutrient conditions in the presence of bacteria were promoted by adding glucose to stimulate bacterial growth and the uptake of N and P released by algal phagotrophy. Subsequent decreases in dissolved N and P following the addition of glucose corresponded to a second period of rapid growth of the alga in both light and dark. This result, combined with evidence for slow axenic growth of this strain, indicated that nutrient acquisition for this species in the presence of bacteria was accomplished primarily via ingestion of bacteria. [ABSTRACT FROM AUTHOR]