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Light availability is the main regulator of primary production, shaping photosynthetic communities and their production of ecologically important biomolecules. In freshwater ecosystems, increasing dissolved organic carbon (DOC) concentrations, commonly known as browning, leads to lower light availability and the proliferation of mixotrophic phytoplankton. Here, a mixotrophic algal species (Cryptomonas sp.) was grown under five increasing DOC concentrations to uncover the plastic responses behind the success of mixotrophs in browning environments and their effect in the availability of nutritionally important biomolecules. In addition to the browning treatments, phototrophic, heterotrophic and mixotrophic growth conditions were used as controls. Despite reduced light availability, browning did not impair algal growth compared to phototrophic conditions. Comparative transcriptomics showed that genes related to photosynthesis were down‐regulated, whereas phagotrophy gene categories (phagosome, lysosome and endocytosis) were up‐regulated along the browning gradient. Stable isotope analysis of phospholipid fractions validated these results, highlighting that the studied mixotroph increases its reliance on heterotrophic processes with browning. Metabolic pathway reconstruction using transcriptomic data suggests that organic carbon is acquired through phagotrophy and used to provide energy in conjunction with photosynthesis. Although metabolic responses to browning were observed, essential fatty acid content was similar between treatments while sterol content was slightly higher upon browning. Together, our results provide a mechanistic model of how a mixotrophic alga responds to browning and how such responses affect the availability of nutritionally essential biomolecules for higher trophic levels. [ABSTRACT FROM AUTHOR]

Production of algae as renewable resources has been pursued actively. Attention has so far been focused on phototrophic algae. Explored here is a completely different approach that harnesses the growth ability of phagotrophic algae on microorganisms such as bacteria, which are abundantly present in waste, including activated sludge generated in wastewater treatment plants. Escherichia coli, a common bacterium, was used as an example of bacterial feed for the oleaginous phagotrophic alga Ochromonas danica. Growth characteristics and lipid contents of O. danica were determined under various conditions. Algal doubling time could be as short as 7 h, but increased to >20 h at low starting E. coli-to- O. danica ratios. The alga was also sensitive to step changes of pH and darkness. Without these factors, phagotrophic growth was similar in a wide range of conditions, even under total darkness and in deionized water with washed E. coli as the only food source. O. danica grew on E. coli with an average yield of about 25% (w/w) and a lipid content of 27% ± 8%. As a food source for phagotrophic algal growth, bacterial cells provided excess nitrogen, which was released as ammonia. Glucose supplementation was studied to avoid ammonia release and produce more algae with higher lipid content (33-34%). Results supported feasibility of this new approach and identified suitable process conditions for further development and application in real wastewater systems. Besides algal production, the approach can remove harmful bacteria from wastewater. [ABSTRACT FROM AUTHOR]

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Lindehoff, Elin, Edna Granéli, and Patricia M. Glibert, 2010. Influence of Prey and Nutritional Status on the Rate of Nitrogen Uptake by Prymnesium parvum (Haptophyte). Journal of the American Water Resources Association (JAWRA) 46(1):121-132. DOI: 10.1111/j.1752-1688.2009.00396.x We studied how the specific nitrogen (N) uptake rates of nitrate (NO3−), urea, and the amino acids, glutamic acid and glycine, by Prymnesium parvum were affected by (1) the change from N-deficient status to N-sufficient status of the P. parvum cells, (2) presence of prey from a natural Baltic Sea plankton community, and (3) the composition of prey as affected by additions of terrestrial originated dissolved organic matter (DOM) or inorganic nutrients. Nitrogen-deficient P. parvum (16 μM NO3− and 4 μM PO4−, molar N:P ratio of 4:1) were mixed with a natural Baltic plankton community and given PO43− and (1) NO3− (control) or (2) high molecular weight DOM, >1 kDa concentrated from sewage effluent (+DOM), in a molar N:P ratio of 9-10:1. With additions of 15N-enriched substrates, rates of N uptake from NO3−, urea, and the amino acids glycine and glutamic acid were measured every 24 h for 72 h. Initial N-deficient P. parvum were highly toxic (3.7 ± 0.9 × 10−4 mg Sap equiv/cell) and toxic allelochemicals were released into the medium causing the natural plankton community to lyse. Rates of N uptake differed between the “control” and the “+DOM” treatments over time; total (sum of the N substrates measured) absolute uptake rates (ρcell, fmol N/cell/h) at ambient culture conditions were significantly higher (ANOVA, p < 0.05) in the more toxic “control” treatments compared with the “+DOM” treatments after 48 h. In the “control” treatment, the total ρcell increased significantly (ANOVA, p < 0.01) from time 0 to 48 h, while in the “+DOM” treatment there was no significant increase. Released organic nutrients from the lysed plankton cells may have increased uptake rates of amino acids and urea by P. parvum. All uptake rates declined in all treatments by 72 h. Total dissolved N uptake rates at ambient culture conditions were estimated to make up about 10% of the N P. parvum are potentially capable of ingesting from particulate prey. [ABSTRACT FROM AUTHOR]

Abstract: Historically most harmful algal species (HAS) have been thought to be strictly phototrophic. Mixotrophy, the use of phototrophy and heterotrophy in combination, has been emphasized as operative mainly in nutrient-poor habitats as a mechanism for augmenting nutrient supplies. Here we examine an alternate premise, that many harmful algae which thrive in eutrophic habitats are mixotrophs that respond both directly to nutrient inputs, and indirectly through high abundance of bacterial and algal prey that are stimulated by the elevated nutrients. From review and synthesis of the available data, mixotrophy occurs in all HAS examined thus far in the organic substrate- and prey-rich habitats of eutrophic estuarine and marine coastal waters. Where data are available comparing phototrophy versus mixotrophy, mixotrophy in eutrophic habitats generally is significant in nutrient acquisition and growth of HAS and, therefore, likely important in the development and maintenance of their blooms. In eutrophic habitats phagotrophic mixotrophs, in particular, have been shown to attain higher growth than when in phototrophic mode. Yet for many HAS, quantitative data about the role of mixotrophy in nutrition, growth, and blooms are lacking, especially relating laboratory information to natural field assemblages, so that the relative importance of photosynthesis, dissolved organic nutrients, and ingestion of prey largely remain unknown. Research is needed to assess simultaneously the roles of phototrophy, osmotrophy and phagotrophy in the nutritional ecology of HAS in eutrophic habitats, spanning bloom initiation, development and senescence. From these data, models that include the role of mixotrophy can be developed to gain more realistic insights about the nutritional factors that control harmful algae in eutrophic waters, and to strengthen predictive capability in predicting their blooms. An overall forecast that can be tested, as well, is that harmful mixotrophic algae will become more abundant as their food supplies increase in many estuaries and coastal waters that are sustaining chronic, increasing cultural eutrophication. [Copyright &y& Elsevier]

Abstract: Deoxyribonuclease II (DNase II) is an endonuclease with optimal activity at low pH, localized within the lysosomes of higher eukaryotes. The origin of this enzyme remains in dispute, and its phylogenetic distribution leaves many questions about its subsequent evolutionary history open. Earlier studies have documented its presence in various metazoans, as well as in Dictyostelium, Trichomonas and, anomalously, a single genus of bacteria (Burkholderia). This study makes use of searches of the genomes of various organisms against known DNase II query sequences, in order to determine the likely point of origin of this enzyme among cellular life forms. Its complete absence from any other bacteria makes prokaryotic origin unlikely. Convincing evidence exists for DNase II homologs in Alveolates such as Paramecium, Heterokonts such as diatoms and water molds, and even tentative matches in green algae. Apparent absences include red algae, plants, fungi, and a number of parasitic organisms. Based on this phylogenetic distribution and hypotheses of eukaryotic relationships, the most probable explanation is that DNase II has been subject to multiple losses. The point of origin is debatable, though its presence in Trichomonas and perhaps in other evolutionarily basal “Excavate” protists such as Reclinomonas, strongly support the hypothesis that DNase II arose as a plesiomorphic trait in eukaryotes. It probably evolved together with phagocytosis, specifically to facilitate DNA degradation and bacteriotrophy. The various absences in many eukaryotic lineages are accounted for by loss of phagotrophic function in intracellular parasites, in obligate autotrophs, and in saprophytes. [Copyright &y& Elsevier]

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]

Cryptoperidiniopsoids are an unclassified group of delicately thecate heterotrophic dinoflagellates known to be common in eastern U.S. estuarine waters. Over the past 10 years cryptoperidiniopsoids were isolated from different geographical regions and cultured with cryptophyte algal prey. In the seven clonal isolates examined, reproduction was strongly linked to the availability of prey cells. The dinoflagellates phagocytized the contents of prey cells through a tube-like peduncle, similarly as close relatives of Pfiesteria spp. and several other heterotrophic species. Cell division occurred while encysted, most commonly yielding two biflagellated offspring. Abundant fusing gametes, phagotrophic planozygotes, and cysts with a pronounced nuclear cyclosis characterized persistent sexuality. Cysts with nuclear cyclosis produced two flagellated offspring cells. The resistance of reproductive cysts to antimicrobial treatments was examined, and a simple high-yield technique was developed for population synchronization while ridding the dinoflagellates of most contaminating vacuolar prey DNA and external contaminants. The DNA content and population DNA profiles of synchronously excysted cryptoperidiniopsoids from different isolates were measured using flow cytometry and were related to the life history of these and other dinoflagellates. Cryptophyte-fed cultures with versus without extracellular bacteria were compared, and bacteria apparently promoted cryptoperidiniopsoid feeding and growth. Externally bacteria-free dinoflagellates were cultured in media enriched with dissolved organic nutrients, and nutritional benefit may have occurred in some treatments. The potential for mixotrophic nutrition from maintenance of cryptophyte chloroplasts was examined using flow cytometrically sorted cells, but evidence of kleptoplastidy was not found in these isolates under the conditions imposed. [ABSTRACT FROM AUTHOR]

Cryptoperidiniopsoids are an unclassified group of delicately thecate heterotrophic dinoflagellates known to be common in eastern U.S. estuarine waters. Over the past 10 years cryptoperidiniopsoids were isolated from different geographical regions and cultured with cryptophyte algal prey. In the seven clonal isolates examined, reproduction was strongly linked to the availability of prey cells. The dinoflagellates phagocytized the contents of prey cells through a tube-like peduncle, similarly as close relatives of Pfiesteria spp. and several other heterotrophic species. Cell division occurred while encysted, most commonly yielding two biflagellated offspring. Abundant fusing gametes, phagotrophic planozygotes, and cysts with a pronounced nuclear cyclosis characterized persistent sexuality. Cysts with nuclear cyclosis produced two flagellated offspring cells. The resistance of reproductive cysts to antimicrobial treatments was examined, and a simple high-yield technique was developed for population synchronization while ridding the dinoflagellates of most contaminating vacuolar prey DNA and external contaminants. The DNA content and population DNA profiles of synchronously excysted cryptoperidiniopsoids from different isolates were measured using flow cytometry and were related to the life history of these and other dinoflagellates. Cryptophyte-fed cultures with versus without extracellular bacteria were compared, and bacteria apparently promoted cryptoperidiniopsoid feeding and growth. Externally bacteria-free dinoflagellates were cultured in media enriched with dissolved organic nutrients, and nutritional benefit may have occurred in some treatments. The potential for mixotrophic nutrition from maintenance of cryptophyte chloroplasts was examined using flow cytometrically sorted cells, but evidence of kleptoplastidy was not found in these isolates under the conditions imposed. [ABSTRACT FROM AUTHOR]

The growth and grazing characteristics of Poterioochromonas malhamensis (Pringsheim) Peterfi (= Ochromonas malhamensis Pringsheim) (ca. 8 μm) feeding on phytoplankton, including the cyanobacteria Synechococcus sp. (ca. 2 μm) and Microcystis viridis (A. Brown) Lemmermann (ca. 6 μm) and the green alga Chlorella pyrenoidosa Chick (ca. 13 μm), were investigated in laboratory experiments involving the following treatments: (1) light without added algal prey (autotrophy), (2) light with added algal prey (mixotrophy), and (3) dark with added algal prey (phagotrophy). There were significantly higher cell numbers under mixotrophic and phagotrophic growth than under autotrophic growth. With phytoplankton as food, growth rates under both mixotrophy and phagotrophy were about two or three times higher than those under autotrophy, indicating that the algal diets were readily able to support the population growth of P. malhamensis. There were no significant differences in growth rate between mixotrophic and phagotrophic cultures during exponential growth. The ingestion rate of P. malhamensis with algal prey was also similar under both continuous light and dark. Poterioochromonas malhamensis ingested on average 0.27 M. viridis cells·flagellate-1·h-1 and 0.18 C. pyrenoidosa cells·flagellate-1·h-1 in continuous light and 0.25 M. viridis cells·flagellate-1·h-1 and 0.18 C. pyrenoidosa cells·flagellate-1·h-1 in continuous dark during exponential growth. The results showed that light had no effect on the growth and ingestion rates of P. malhamensis for phagotrophy during exponential growth. However, phagotrophic populations of P. malhamensis were incapable of growth in continuous darkness for longer than 5 days. Populations of P. malhamensis showed no increase when prey was added again after 4 days in continuous darkness, indicating that... [ABSTRACT FROM AUTHOR]

Sulcochrysis biplastida gen. et sp. nov., a golden, marine, mixotrophic flagellate is described. Cells resemble Ochromonas in light microscopic features, but they are distinct at the electron microscopic level from Ochromonas or any other typical chrysophyte. Ultrastructural features that discriminate Sulcochrysis from the Chrysophyceae are: (i) a proximal helix in the flagellar transition region; (ii) basal bodies situated in the anterior depression of the nucleus; (iii) the lack of the rhizoplast; and (iv) simple flagellar hairs lacking lateral filaments. These features suggest that Sulcochrysis is a relative of the Pedinellophyceae, Dictyochophyceae and Pelagophyceae. However, Sulcochrysis has a flagellar root system similar to that of the Ochromonas-type cell and it may use the R3 root for prey capture, as do ochromonadalean algae. The R3 root and the phagotrophic mechanism using the R3 root are interpreted as a plesiomorphy, because these are also distributed in primitive heterokonts such as the bicosoecids. Sulcochrysis has one microtubule probably homologous with the x-fibre of Bicosoeca maris Pic ken. Based on these features it is suggested that Sulcochrysis is an organism that links bicosoecids (Bicosoecophyceae), Pedinellophyceae, Dictyochophyceae and Pelagophyceae. [ABSTRACT FROM AUTHOR]