A biotic video game smart phone kit for formal and informal biophysics education

NASA Astrophysics Data System (ADS)

Kim, Honesty; Lee, Seung Ah; Riedel-Kruse, Ingmar

2015-03-01

Novel ways for formal and informal biophysics education are important. We present a low-cost biotic game design kit that incorporates microbial organisms into an interactive gaming experience: A 3D-printable microscope containing four LEDs controlled by a joystick enable human players to provide directional light stimuli to the motile single-celled organism Euglena gracilis. These cellular behaviors are displayed on the integrated smart phone. Real time cell-tracking couples these cells into interactive biotic video game play, i.e., the human player steers Euglena to play soccer with virtual balls and goals. The player's learning curve in mastering this fun game is intrinsically coupled to develop a deeper knowledge about Euglena's cell morphology and the biophysics of its phototactic behavior. This kit is dual educational - via construction and via play - and it provides an engaging theme for a formal biophysics devices class as well as to be presented in informal outreach activities; its low cost and open soft- and hardware should enable wide adoption.

Impact of a high magnetic field on the orientation of gravitactic unicellular organisms--a critical consideration about the application of magnetic fields to mimic functional weightlessness.

PubMed

Hemmersbach, Ruth; Simon, Anja; Waßer, Kai; Hauslage, Jens; Christianen, Peter C M; Albers, Peter W; Lebert, Michael; Richter, Peter; Alt, Wolfgang; Anken, Ralf

2014-03-01

The gravity-dependent behavior of Paramecium biaurelia and Euglena gracilis have previously been studied on ground and in real microgravity. To validate whether high magnetic field exposure indeed provides a ground-based facility to mimic functional weightlessness, as has been suggested earlier, both cell types were observed during exposure in a strong homogeneous magnetic field (up to 30 T) and a strong magnetic field gradient. While swimming, Paramecium cells were aligned along the magnetic field lines; orientation of Euglena was perpendicular, demonstrating that the magnetic field determines the orientation and thus prevents the organisms from the random swimming known to occur in real microgravity. Exposing Astasia longa, a flagellate that is closely related to Euglena but lacks chloroplasts and the photoreceptor, as well as the chloroplast-free mutant E. gracilis 1F, to a high magnetic field revealed no reorientation to the perpendicular direction as in the case of wild-type E. gracilis, indicating the existence of an anisotropic structure (chloroplasts) that determines the direction of passive orientation. Immobilized Euglena and Paramecium cells could not be levitated even in the highest available magnetic field gradient as sedimentation persisted with little impact of the field on the sedimentation velocities. We conclude that magnetic fields are not suited as a microgravity simulation for gravitactic unicellular organisms due to the strong effect of the magnetic field itself, which masks the effects known from experiments in real microgravity.

Long term stability of Oligo (dT) 25 magnetic beads for the expression analysis of Euglena gracilis for long term space projects

NASA Astrophysics Data System (ADS)

Becker, Ina; Strauch, Sebastian M.; Hauslage, Jens; Lebert, Michael

2017-05-01

The unicellular freshwater flagellate Euglena gracilis has a highly developed sensory system. The cells use different stimuli such as light and gravity to orient themselves in the surrounding medium to find areas for optimal growth. Due to the ability to produce oxygen and consume carbon dioxide, Euglena is a suitable candidate for life support systems. Participation in a long-term space experiment would allow for the analysis of changes and adaptations to the new environment, and this could bring new insights into the mechanism of perception of gravity and the associated signal transduction chain. For a molecular analysis of transcription patterns, an automated system is necessary, capable of performing all steps from taking a sample, processing it and generating data. One of the developmental steps is to find long-term stable reagents and materials and test them for stability at higher-than-recommended temperature conditions during extended storage time. We investigated the usability of magnetic beads in an Euglena specific lysis buffer after addition of the RNA stabilizer Dithiothreitol over 360 days and the lysis buffer with the stabilizer alone over 455 days at the expected storage temperature of 19 °C. We can claim that the stability is not impaired at all after an incubation period of over one year. This might be an interesting result for researchers who have to work under non-standard lab conditions, as in biological or medicinal fieldwork.

Aliphatic Chains of Esterified Lipids in Isolated Eyespots of Euglena gracilis var. bacillaris1

PubMed Central

Hilenski, Lula L.; Walne, Patricia L.; Snyder, Fred

1976-01-01

Isolated eyespot granules of Euglena gracilis Klebs var. bacillaris Pringsheim contained approximately 6% lipids (based on protein). Separation of the lipid extracts by thin layer chromatography revealed four major fractions: wax esters, triacylglycerols, free fatty acids, and phospholipids. Methanolysis of each fraction yielded between 27 and 29 different fatty acids ranging from 12:0 to 22:6. Acetates of the fatty alcohols of the wax fraction consisted of 11:0 to 18:0 carbon chains, with 14:0 being the major component; unsaturated alcohols were not detected. PMID:16659543

Gravitaxis in Euglena.

PubMed

Häder, Donat-P; Hemmersbach, Ruth

2017-01-01

Motile microorganisms utilize a number of responses to external stimuli including light, temperature, chemicals as well as magnetic and electric fields. Gravity is a major clue to select a niche in their environment. Positive gravitaxis leads an organism down into the water column and negative gravitaxis brings it to the surface. In Euglena the precision of gravitaxis is regulated by an internal rhythm entrained by the daily light/dark cycle. This and the cooperation with phototaxis bring the cells into an optimal position in the water column. In the past a passive orientation based on a buoy mechanism has been proposed for Euglena gracilis, but now it has been proven that this flagellate possesses a physiological gravireceptor and an active orientation. Numerous experiments in space using satellites, rockets and shuttles as well as in parabolic flights have been conducted as well as in functional weightlessness (simulated microgravity) on ground-based facilities such as clinostats to characterize the gravitaxis of Euglena. The threshold for gravity perception was determined and physiological, biochemical and molecular components of the signal transduction chain have been identified. In contrast to higher plants, some algae and ciliates, Euglena does not possess sedimenting statoliths to detect the direction of the gravity vector of the Earth. The gravireceptors were found to be mechano-sensitive Ca 2+ -conducting ion channels thought to be located at the front end of the cell underneath the trailing flagellum. When activated by gravity-induced pressure due to sedimentation of the whole cell body, they allow a passive influx of calcium along a previously established ion gradient. The entering calcium binds to a specific calmodulin (CaM.2) which in turn activates an adenylyl cyclase producing cAMP from ATP. This cAMP is believed to activate a specific protein kinase A (PK.4), which is postulated to phosphorylate proteins inside the flagellum resulting in a bending and thus a course correction and reorientation with respect to the direction of the gravity vector. The elements of the signal transduction chain have been characterized by inhibitors and by RNAi to prove their involvement in gravitaxis.

Evolutionary appearance of genes encoding proteins associated with box H/ACA snoRNAs: Cbf5p in Euglena gracilis, an early diverging eukaryote, and candidate Gar1p and Nop10p homologs in archaebacteria

PubMed Central

Watanabe, Yoh-ichi; Gray, Michael W.

2000-01-01

A reverse transcription–polymerase chain reaction (RT–PCR) approach was used to clone a cDNA encoding the Euglena gracilis homolog of yeast Cbf5p, a protein component of the box H/ACA class of snoRNPs that mediate pseudouridine formation in eukaryotic rRNA. Cbf5p is a putative pseudouridine synthase, and the Euglena homolog is the first full-length Cbf5p sequence to be reported for an early diverging unicellular eukaryote (protist). Phylogenetic analysis of putative pseudouridine synthase sequences confirms that archaebacterial and eukaryotic (including Euglena) Cbf5p proteins are specifically related and are distinct from the TruB/Pus4p clade that is responsible for formation of pseudouridine at position 55 in eubacterial (TruB) and eukaryotic (Pus4p) tRNAs. Using a bioinformatics approach, we also identified archaebacterial genes encoding candidate homologs of yeast Gar1p and Nop10p, two additional proteins known to be associated with eukaryotic box H/ACA snoRNPs. These observations raise the possibility that pseudouridine formation in archaebacterial rRNA may be dependent on analogs of the eukaryotic box H/ACA snoRNPs, whose evolutionary origin may therefore predate the split between Archaea (archaebacteria) and Eucarya (eukaryotes). Database searches further revealed, in archaebacterial and some eukaryotic genomes, two previously unrecognized groups of genes (here designated ‘PsuX’ and ‘PsuY’) distantly related to the Cbf5p/TruB gene family. PMID:10871366

Long term stability of Oligo (dT) 25 magnetic beads for the expression analysis of Euglena gracilis for long term space projects.

PubMed

Becker, Ina; Strauch, Sebastian M; Hauslage, Jens; Lebert, Michael

2017-05-01

The unicellular freshwater flagellate Euglena gracilis has a highly developed sensory system. The cells use different stimuli such as light and gravity to orient themselves in the surrounding medium to find areas for optimal growth. Due to the ability to produce oxygen and consume carbon dioxide, Euglena is a suitable candidate for life support systems. Participation in a long-term space experiment would allow for the analysis of changes and adaptations to the new environment, and this could bring new insights into the mechanism of perception of gravity and the associated signal transduction chain. For a molecular analysis of transcription patterns, an automated system is necessary, capable of performing all steps from taking a sample, processing it and generating data. One of the developmental steps is to find long-term stable reagents and materials and test them for stability at higher-than-recommended temperature conditions during extended storage time. We investigated the usability of magnetic beads in an Euglena specific lysis buffer after addition of the RNA stabilizer Dithiothreitol over 360 days and the lysis buffer with the stabilizer alone over 455 days at the expected storage temperature of 19 °C. We can claim that the stability is not impaired at all after an incubation period of over one year. This might be an interesting result for researchers who have to work under non-standard lab conditions, as in biological or medicinal fieldwork. Copyright © 2017 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.

Impact of a High Magnetic Field on the Orientation of Gravitactic Unicellular Organisms—A Critical Consideration about the Application of Magnetic Fields to Mimic Functional Weightlessness

PubMed Central

Simon, Anja; Waßer, Kai; Hauslage, Jens; Christianen, Peter C.M.; Albers, Peter W.; Lebert, Michael; Richter, Peter; Alt, Wolfgang; Anken, Ralf

2014-01-01

Abstract The gravity-dependent behavior of Paramecium biaurelia and Euglena gracilis have previously been studied on ground and in real microgravity. To validate whether high magnetic field exposure indeed provides a ground-based facility to mimic functional weightlessness, as has been suggested earlier, both cell types were observed during exposure in a strong homogeneous magnetic field (up to 30 T) and a strong magnetic field gradient. While swimming, Paramecium cells were aligned along the magnetic field lines; orientation of Euglena was perpendicular, demonstrating that the magnetic field determines the orientation and thus prevents the organisms from the random swimming known to occur in real microgravity. Exposing Astasia longa, a flagellate that is closely related to Euglena but lacks chloroplasts and the photoreceptor, as well as the chloroplast-free mutant E. gracilis 1F, to a high magnetic field revealed no reorientation to the perpendicular direction as in the case of wild-type E. gracilis, indicating the existence of an anisotropic structure (chloroplasts) that determines the direction of passive orientation. Immobilized Euglena and Paramecium cells could not be levitated even in the highest available magnetic field gradient as sedimentation persisted with little impact of the field on the sedimentation velocities. We conclude that magnetic fields are not suited as a microgravity simulation for gravitactic unicellular organisms due to the strong effect of the magnetic field itself, which masks the effects known from experiments in real microgravity. Key Words: Levitation—Microgravity—Gravitaxis—Gravikinesis—Gravity. Astrobiology 14, 205–215. PMID:24621307

Measurements of Euglena motion parameters by laser light scattering.

PubMed Central

Ascoli, C; Barbi, M; Frediani, C; Murè, A

1978-01-01

Measurements of Euglena gracilis motion parameters have been performed by the spectral analysis of the scattered laser light. Samples were oriented by a radiofrequency field to obtain easily interpretable spectra. Cell rotation frequency and flagellar beating frequency distributions were obtained from the homodyne spectra, whereas the Doppler lines obtained at small observation angles by heterodyne detection yielded the swimming speed distributions. We discuss the broadening of the heterodyne spectra at large angles of observation. An application of this method to the study of the photo-kinetic effect is also described. Images FIGURE 3 PMID:104747

Applicability of Euglena gracilis for biorefineries demonstrated by the production of α-tocopherol and paramylon followed by anaerobic digestion.

PubMed

Grimm, Philipp; Risse, Joe M; Cholewa, Dominik; Müller, Jakob M; Beshay, Usama; Friehs, Karl; Flaschel, Erwin

2015-12-10

In this study the use of Euglena gracilis biomass for α-tocopherol, paramylon and biogas production in a value-added chain was investigated. Therefore, we analyzed the dry cell weight and product concentrations at different growth phases during heterotrophic, photoheterotrophic and photoautotrophic cultivation in a low-cost minimal medium. Furthermore, the specific biogas yields for differently derived biomass with and without product recovery were investigated. We demonstrate that growth phase and cultivation mode not only have a significant impact on product formation, but also influence the yield of biogas obtained from anaerobic digestion of Euglena gracilis biomass. The maximum dry cell weight concentration ranged from 12.3±0.14gL(-1) for heterotrophically to 3.4±0.02gL(-1) for photoautotrophically grown Euglena gracilis cells. The heterotrophically grown biomass accumulated product concentrations of 5.3±0.12mgL(-1) of α-tocopherol and 9.3±0.1gL(-1) of paramylon or 805±10.9mL of biogasgvs(-1) (per gram volatile solids). The results for photoautotrophically grown cells were 8.6±0.22mgL(-1) of α-tocopherol and 0.78±0.01gL(-1) of paramylon or 648±7.2mL of biogasgvs(-1). For an energy-saving downstream procedure the extracting agent methanol does not have to be removed strictly. Samples with residual methanol showed a significantly increased biogas yield, because the solvent can be used as an additional substrate for methane production by archaebacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

[Ultrastructural description of Euglena pailasensis (Euglenozoa) from Rincón de la Vieja volcano, Guanacaste, Costa Rica].

PubMed

Sánchez, Ethel; Vargas, Maribel; Mora, Marielos; Ortega, José Maria; Serrano, Aurelio; Freer, Enrique; Sittenfeld, Ana

2004-03-01

The euglenoids are unicellular eukaryotic flagellates living in a diversity of soils and aquatic environments and ecosystems. This study describes the ultrastructure of an euglenoid isolated from the surface of a boiling mud pool with temperatures ranging from 38 to 98 degrees C and pH 2 - 4. The hot mud pool is located in Area de Pailas de Barro, Las Pailas, Rincón de la Vieja Volcano, Guanacaste, Costa Rica. The morphological characterization of the Euglena pailasensis was performed by SEM and TEM. It was determined that, although the euglenoid was obtained from an extreme volcanic environment, the general morphology corresponds to that of a typical member of Euglena of 30-45 microm long and 8-10 microm wide, with membrane, pellicle, chloroplasts, mitochondria, nucleus, pigments and other cytoplasmic organelles. E. pailasensis is delimited by a membrane and by 40 to 90 pellicle strips. It was observed up to 5 elongated chloroplasts per cell. The chloroplast contains several osmiophilic globules and a pyrenoid penetrated by few thylakoid pairs. The nutritious material is reserved in numerous small paramylon grains located at the center of the cell, mitocondria are characterized by the presence of crests in radial disposition toward the interior of the lumen. It was also observed around the external surface "pili" like filaments originating from the pellicle strips. There is no evidence for the presence of flagella in the ampulla (reservoir/canal area), a fact confirmed by negative staining, and a difference regarding other species of Euglena. The observed ultrastructural characteristics are not sufficient to explain the adaptation of this species to acid and hot environments.

Different effects of eubacterial and eukaryotic DNA topoisomerase II inhibitors on chloroplasts ofEuglena gracilis

NASA Astrophysics Data System (ADS)

Krajčovič, Juraj; Ebringer, Libor

1990-03-01

Inhibitors of eubacterial and eukaryotic DNA topoisomerases type II exhibited different effects on chloroplasts of the flagellateEuglena gracilis. Antibacterial agents (cinoxacin, nalidixic and oxolinic acids, ciprofloxacin, enoxacin, norfloxacin and ofloxacin) from the group of quinolones and coumarins (coumermycin A1, clorobiocin and novobiocin) — all inhibitors of prokaryotic DNA topoisomerase II — were very potent eliminators of chloroplasts fromE. gracilis. In contrast, antitumor drugs (adriamycin, etoposide, teniposide and mitoxantrone) — antagonists of the eukaryotic counterpart — did not affect these semiautonomous photosynthetic organelles. These findings point out again the close evolutionary relationships between eubacteria and chloroplasts and are in agreement with the hypothesis of an endosymbiotic origin of chloroplasts.

Critical Involvement of Environmental Carbon Dioxide Fixation to Drive Wax Ester Fermentation in Euglena

PubMed Central

Nishio, Kazuki; Nakazawa, Masami; Nakamoto, Masatoshi; Okazawa, Atsushi; Kanaya, Shigehiko; Arita, Masanori

2016-01-01

Accumulation profiles of wax esters in Euglena gracilis Z were studied under several environmental conditions. The highest amount of total wax esters accumulated under hypoxia in the dark, and C28 (myristyl-myristate, C14:0-C14:0) was prevalent among all conditions investigated. The wax ester production was almost completely suppressed under anoxia in the light, and supplying exogenous inorganic carbon sources restored wax ester fermentation, indicating the need for external carbon sources for the wax ester fermentation. 13C-labeling experiments revealed specific isotopic enrichment in the odd-numbered fatty acids derived from wax esters, indicating that the exogenously-supplied CO2 was incorporated into wax esters via the propionyl-CoA pathway through the reverse tricarboxylic acid (TCA) cycle. The addition of 3-mercaptopicolinic acid, a phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, significantly affected the incorporation of 13C into citrate and malate as the biosynthetic intermediates of the odd-numbered fatty acids, suggesting the involvement of PEPCK reaction to drive wax ester fermentation. Additionally, the 13C-enrichment pattern of succinate suggested that the CO2 assimilation might proceed through alternative pathways in addition to the PEPCK reaction. The current results indicate that the mechanisms of anoxic CO2 assimilation are an important target to reinforce wax ester fermentation in Euglena. PMID:27669566

Restart capability of resting-states of Euglena gracilis after 9 months of dormancy: preparation for autonomous space flight experiments

NASA Astrophysics Data System (ADS)

Strauch, Sebastian M.; Becker, Ina; Pölloth, Laura; Richter, Peter R.; Haag, Ferdinand W. M.; Hauslage, Jens; Lebert, Michael

2018-04-01

Dormant states of organisms are easier to store than the living state because they tolerate larger variations in temperature, light, storage space etc., making them attractive for laboratory culture stocks and also for experiments under special circumstances, especially space flight experiments. Like several other organisms, Euglena gracilis is capable of forming desiccation tolerant resting states in order to survive periods of unfavourable environmental conditions. In earlier experiments it was found that dormant Euglena cells must not become completely desiccated. Some residual moisture is required to ensure recovery of the resting states. To analyse the water demand in recovery of Euglena resting states, cells were transferred to a defined amount of cotton wool (0.5 g). Subsequently different volumes of medium (1, 2, 3, 4, 5, 8, 10 and 20 ml) were added in order to supply humidity; a control was set up without additional liquid. Samples were sealed in transparent 50 ml falcon tubes and stored for 9 months under three different conditions: • Constant low light conditions in a culture chamber at 20°C, • In a black box, illuminated with short light emitting diode-light pulses provided by joule thieves and • In darkness in a black box. After 9 months, cells were transferred to fresh medium and cell number, photosynthetic efficiency and movement behavior was monitored over 3 weeks. It was found that cells recovered under all conditions except in the control, where no medium was supplied. Transcription levels of 21 genes were analysed with a Multiplex-polymerase chain reaction. One hour after rehydration five of these genes were found to be up-regulated: ubiquitin, heat shock proteins HSP70, HSP90, the calcium-sensor protein frequenin and a distinct protein kinase, which is involved in gravitaxis. The results indicate a transient general stress response of the cells.

Comparative assessment of the Euglena gracilis var. saccharophila variant strain as a producer of the β-1,3-glucan paramylon under varying light conditions.

PubMed

Sun, Angela; Hasan, Mafruha Tasnin; Hobba, Graham; Nevalainen, Helena; Te'o, Junior

2018-06-11

Euglena gracilis Z and a 'sugar loving' variant strain Euglena gracilis var. saccharophila, were investigated as producers of paramylon, a β-1,3-glucan polysaccharide with potential medicinal and industrial applications. The strains were grown under diurnal or dark growth conditions on a glucose-yeast extract medium supporting high-level paramylon production. Both strains produced the highest paramylon yields (7.4 to 8 g · L -1 , respectively) while grown in the dark, but the maximum yield was achieved faster by E. gracilis var. saccharophila (48 h vs 72 h). The glucose-to-paramylon yield coefficient Y par/glu = 0.46±0.03 in the Euglena gracilis var. saccharophila cultivation, obtained in this study, is the highest reported to date. Proteomic analysis of the metabolic pathways provided molecular clues for the strain behavior observed during cultivation. For example, overexpression of enzymes in the gluconeogenesis/glycolysis pathways including fructokinase-1 and chloroplastic fructose-1,6-bisphosphatase may have contributed to the faster rate of paramylon accumulation in E. gracilis var. saccharophila. Differentially expressed proteins in the early steps of chloroplastogenesis pathway including plastid uroporphyrinogen decarboxylases, photoreceptors, and a highly abundant (68 fold increase) plastid transketolase may have provided the E. gracilis var. saccharophila strain an advantage in paramylon production during diurnal cultivations. In conclusion, the variant strain E. gracilis var. saccharophila seems to be well suited for producing large amounts of paramylon. This work has also resulted in the identification of molecular targets for future improvement of paramylon production in E. gracilis, including the chloroplastic fructose-1,6-bisphosphatase (FBP) and phosophofructokinase 1 (PFK-1), the latter being a key regulator of glycolysis. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Motility and gravitactic orientation of the flagellate, Euglena gracilis, impaired by artificial and solar UV-B radiation.

PubMed

Hader, D P; Liu, S M

1990-09-01

The effects of ultraviolet radiation on the gravitactic orientation of the freshwater flagellate, Euglena gracilis, were determined by a real time image analysis system. Both artificial UV radiation and solar radiation in a temperature-controlled growth chamber were employed. Histograms of gravitaxis showed that the degree of orientation decreased with increasing exposure time; this can be quantified using the Rayleigh test and upper quadrant summation. The effects of artificial UV radiation on the orientation are considerably stronger than those of solar radiation, probably because the radiation source emits higher fluence rates below 300 nm than found in solar radiation. The effects of monochromatic ultraviolet radiation on motility have been determined, and an action spectrum has been calculated.

Modular 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms

NASA Astrophysics Data System (ADS)

Podwin, Agnieszka; Dziuban, Jan A.

2017-10-01

The paper presents the sandwiched polymer 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms. Euglenas and yeast were separately and simultaneously cultured for 10 d in the chip. As a result of the experiments, euglenas, light-initialized and nourished by CO2—a product of ethanol fermentation handled by yeast—generated oxygen, based on the photosynthesis process. The presence of oxygen in the bio-reactor was confirmed by the colorimetric method—a bicarbonate (pH) indicator. Preliminary studies towards the obtainment of an effective source of oxygen are promising and further research should be done to enable the utility of the bio-reactor in, for instance, microbial fuel cells.

Genotoxicity and subchronic toxicity evaluation of dried Euglena gracilis ATCC PTA-123017.

PubMed

Simon, Ryan R; Vo, Trung D; Levine, Robert

2016-10-01

Euglena gracilis is a microalga capable of synthesizing various nutrients of interest in human and animal nutrition. When cultivated aerobically in the dark, Euglena synthesize paramylon, a storage polysaccharide comprised of high molecular weight beta-1,3-D-glucose polymers organized in cytoplasmic granules. Beta-glucans have been shown to have immune modulation effects, including anti-microbial, anti-tumor, and anti-oxidant properties, and metabolic effects, such as regulation of cholesterol and blood sugar levels. Preparations of E. gracilis and paramylon may therefore have potential utility as functional food ingredients for human and animal nutrition. A battery of toxicological studies was conducted on a dried preparation of E. gracilis and paramylon to support their safe food use. The dried alga was not genotoxic in a bacterial reverse mutation test and mammalian micronucleus test. In the subchronic toxicity study, rats were provided E. gracilis in the diet at levels of 0, 12,500, 25,000 or 50,000 ppm. Paramylon was provided at a concentration of 50,000 ppm. No effects that could be attributable to treatment were observed in clinical observations, body weight, food consumption, ophthalmology, hematology and clinical chemistry, urinalysis, and macroscopic and microscopic findings. A NOAEL of 50,000 ppm in the diet was determined for both ingredients. Copyright © 2016 Elsevier Inc. All rights reserved.

RuBisCO in Non-Photosynthetic Alga Euglena longa: Divergent Features, Transcriptomic Analysis and Regulation of Complex Formation.

PubMed

Záhonová, Kristína; Füssy, Zoltán; Oborník, Miroslav; Eliáš, Marek; Yurchenko, Vyacheslav

2016-01-01

Euglena longa, a close relative of the photosynthetic model alga Euglena gracilis, possesses an enigmatic non-photosynthetic plastid. Its genome has retained a gene for the large subunit of the enzyme RuBisCO (rbcL). Here we provide new data illuminating the putative role of RuBisCO in E. longa. We demonstrated that the E. longa RBCL protein sequence is extremely divergent compared to its homologs from the photosynthetic relatives, suggesting a possible functional shift upon the loss of photosynthesis. Similarly to E. gracilis, E. longa harbors a nuclear gene encoding the small subunit of RuBisCO (RBCS) as a precursor polyprotein comprising multiple RBCS repeats, but one of them is highly divergent. Both RBCL and the RBCS proteins are synthesized in E. longa, but their abundance is very low compared to E. gracilis. No RBCS monomers could be detected in E. longa, suggesting that processing of the precursor polyprotein is inefficient in this species. The abundance of RBCS is regulated post-transcriptionally. Indeed, blocking the cytoplasmic translation by cycloheximide has no immediate effect on the RBCS stability in photosynthetically grown E. gracilis, but in E. longa, the protein is rapidly degraded. Altogether, our results revealed signatures of evolutionary degradation (becoming defunct) of RuBisCO in E. longa and suggest that its biological role in this species may be rather unorthodox, if any.

Effects of temperature, CO2/O2 concentrations and light intensity on cellular multiplication of microalgae, Euglena gracilis

NASA Technical Reports Server (NTRS)

Kitaya, Y.; Azuma, H.; Kiyota, M.

2005-01-01

Microalgae culture is likely to play an important role in aquatic food production modules in bioregenerative systems for producing feeds for fish, converting CO2 to O2 and remedying water quality as well as aquatic higher plants. In the present study, the effects of culture conditions on the cellular multiplication of microalgae, Euglena gracilis, was investigated as a fundamental study to determine the optimum culture conditions for microalgae production in aquatic food production modules including both microalgae culture and fish culture systems. E. gracilis was cultured under conditions with five levels of temperatures (25-33 degrees C), three levels of CO2 concentrations (2-6%), five levels of O2 concentrations (10-30%), and six levels of photosynthetic photon flux (20-200 micromoles m-2 s-1). The number of Euglena cells in a certain volume of solution was monitored with a microscope under each environmental condition. The multiplication rate of the cells was highest at temperatures of 27-31 degrees C, CO2 concentration of 4%, O2 concentration of 20% and photosynthetic photon flux of about 100 micromoles m-2 s-1. The results demonstrate that E. gracilis could efficiently produce biomass and convert CO2 to O2 under relatively low light intensities in aquatic food production modules. c2005 Published by Elsevier Ltd on behalf of COSPAR.

Euglena gracilis chloroplast DNA: analysis of a 1.6 kb intron of the psb C gene containing an open reading frame of 458 codons.

PubMed

Montandon, P E; Vasserot, A; Stutz, E

1986-01-01

We retrieved a 1.6 kbp intron separating two exons of the psb C gene which codes for the 44 kDa reaction center protein of photosystem II. This intron is 3 to 4 times the size of all previously sequenced Euglena gracilis chloroplast introns. It contains an open reading frame of 458 codons potentially coding for a basic protein of 54 kDa of yet unknown function. The intron boundaries follow consensus sequences established for chloroplast introns related to class II and nuclear pre-mRNA introns. Its 3'-terminal segment has structural features similar to class II mitochondrial introns with an invariant base A as possible branch point for lariat formation.

One-Celled Creatures From the Black Lagoon

ERIC Educational Resources Information Center

McClellan, Ray E.

1974-01-01

Describes methods of developing and using cultures of microoganisms suitable for use in elementary school science classes. Emphasis is placed upon the characteristics and culturing of amoeba, paramecia, euglena, rotifers, and algae. (JR)

Microscopic Gardens: A Close Look at Algae.

ERIC Educational Resources Information Center

Foote, Mary Ann

1983-01-01

Describes classroom activities using algae, including demonstration of eutrophication, examination of mating strains, and activities with Euglena. Includes on algal morphology/physiology, types of algae, and field sources for collecting these organisms. (JN)

Kinematics of flagellar swimming in Euglena gracilis: Helical trajectories and flagellar shapes.

PubMed

Rossi, Massimiliano; Cicconofri, Giancarlo; Beran, Alfred; Noselli, Giovanni; DeSimone, Antonio

2017-12-12

The flagellar swimming of euglenids, which are propelled by a single anterior flagellum, is characterized by a generalized helical motion. The 3D nature of this swimming motion, which lacks some of the symmetries enjoyed by more common model systems, and the complex flagellar beating shapes that power it make its quantitative description challenging. In this work, we provide a quantitative, 3D, highly resolved reconstruction of the swimming trajectories and flagellar shapes of specimens of Euglena gracilis We achieved this task by using high-speed 2D image recordings taken with a conventional inverted microscope combined with a precise characterization of the helical motion of the cell body to lift the 2D data to 3D trajectories. The propulsion mechanism is discussed. Our results constitute a basis for future biophysical research on a relatively unexplored type of eukaryotic flagellar movement. Copyright © 2017 the Author(s). Published by PNAS.

Chronic toxicity of a laundry detergent to the freshwater flagellate Euglena gracilis.

PubMed

Azizullah, Azizullah; Richter, Peter; Jamil, Muhammad; Häder, Donat-Peter

2012-10-01

Chronic toxicity of the common laundry detergent Ariel on the freshwater alga Euglena gracilis was investigated by growing the alga in a medium containing the detergent for 7 days. Cell density, motility, swimming velocity, gravitactic orientation, cell shape, photosynthesis and concentration of light-harvesting pigments were used as end point parameters for the assessment of toxicity. Cell density was significantly reduced at a concentration of 1 mg l(-1) or above. Among the other tested parameters, with the exception of cell shape, gravitaxis and chlorophyll b, all were adversely affected by the detergent at concentrations exceeding 1 mg l(-1). It is concluded that long-term (7-days) exposure to the detergent caused significant toxicity to E. gracilis. Furthermore, long-term tests with E. gracilis can be used as sensitive indicator for the toxicity assessment of laundry detergents in aquatic environments.

Aquacells — Flagellates under long-term microgravity and potential usage for life support systems

NASA Astrophysics Data System (ADS)

Häder, Donat-P.; Richter, Peter R.; Strauch, S. M.; Schuster, M.

2006-09-01

The motile behavior of the unicellular photosynthetic flagellate Euglena gracilis was studied during a two-week mission on the Russian satellite Foton M2. The precision of gravitactic orientation was high before launch and, as expected, the cells were unoriented during microgravity. While after previous short-term TEXUS flights the precision of orientation was as high as before launch, it took several hours for the organisms to regain their gravitaxis. Also the percentage of motile cells and the swimming velocity of the remaining motile cells were considerably lower than in the ground control. In preparatory experiments the flagellate Euglena was shown to produce considerable amounts of photosynthetically generated oxygen. In a coupling experiment in a prototype for a planned space mission on Foton M3, the photosynthetic producers were shown to supply sufficient amounts of oxygen to a fish compartment with 35 larval cichlids, Oreochromis mossambicus.

A Closed Ecological System in a Space Experiment

NASA Astrophysics Data System (ADS)

Strauch, S. M.; Schuster, M.; Lebert, M.; Richter, P.; Schmittnagel, M.; Hader, D.-P.

2008-06-01

The Russian FOTON-M3 mission, a satellite for mid-length experiments in space and recovery afterwards, included a closed artificial ecosystem (OMEGAHAB for Oreochromis Mossambicus-Euglena Gracilis-Aquatic HABitat) with the photosynthetic flagellate Euglena gracilis as oxygen producer and larvae of Oreochromis mossambicus, a Tilapia species, as consumer. During the 12-day orbital flight the algae were observed 10 minutes per day by means of a miniaturized microscope to analyse their swimming behavior. The fishes were also filmed to monitor their development and movement. An identical experiment was carried out as ground control. A data downlink provided the measured temperature values of the space experiment every day to readjust the temperature of the ground reference in order to eliminate the influence of the different temperature on the velocity of the development of the fishes. The system worked very well and confirmed the design in principle. OMEGAHAB was the most successful German experiment of that kind as yet.

High-throughput optofluidic profiling of Euglena gracilis with morphological and chemical specificity

NASA Astrophysics Data System (ADS)

Guo, Baoshan; Lei, Cheng; Ito, Takuro; Jiang, Yiyue; Ozeki, Yasuyuki; Goda, Keisuke

2016-11-01

The world is faced with environmental problems and the energy crisis due to the combustion and depletion of fossil fuels. The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, algal biofuel is expected to play a key role in alleviating global warming since algae absorb atmospheric CO2 via photosynthesis. Among various algae for fuel production, Euglena gracilis is an attractive microalgal species as it is known to produce wax ester (good for biodiesel and aviation fuel) within lipid droplets. To date, while there exist many techniques for inducing microalgal cells to produce and accumulate lipid with high efficiency, few analytical methods are available for characterizing a population of such lipid-accumulated microalgae including E. gracilis with high throughout, high accuracy, and single-cell resolution simultaneously. Here we demonstrate a high-throughput optofluidic Euglena gracilis profiler which consists of an optical time-stretch microscope and a fluorescence analyzer on top of an inertial-focusing microfluidic device that can detect fluorescence from lipid droplets in their cell body and provide images of E. gracilis cells simultaneously at a high throughput of 10,000 cells/s. With the multi-dimensional information acquired by the system, we classify nitrogen-sufficient (ordinary) and nitrogen-deficient (lipid-accumulated) E. gracilis cells with a low false positive rate of 1.0%. This method provides a promise for evaluating the efficiency of lipid-inducing techniques for biofuel production, which is also applicable for identifying biomedical samples such as blood cells and cancer cells.

The influence of microgravity on Euglena gracilis as studied on Shenzhou 8.

PubMed

Nasir, A; Strauch, S M; Becker, I; Sperling, A; Schuster, M; Richter, P R; Weißkopf, M; Ntefidou, M; Daiker, V; An, Y A; Li, X Y; Liu, Y D; Lebert, M

2014-01-01

The German Aerospace Center (DLR) enabled German participation in the joint space campaign on the unmanned Shenzhou 8 spacecraft in November 2011. In this report, the effect of microgravity on Euglena gracilis cells is described. Custom-made dual compartment cell fixation units (containing cells in one chamber and fixative - RNA lysis buffer - in another one) were enclosed in a small container and placed in the Simbox incubator, which is an experiment support system. Cells were fixed by injecting them with fixative at different time intervals. In addition to stationary experiment slots, Simbox provides a 1 g reference centrifuge. Cell fixation units were mounted in microgravity and 1 g reference positions of Simbox. Two Simbox incubators were used, one for space flight and the other as ground reference. Cells were fixed soon after launch and shortly before return of the spaceship. Due to technical problems, only early in-flight samples (about 40 min after launch microgravity and corresponding 1 g reference) were fully mixed with fixative, therefore only data from those samples are presented. Transcription of several genes involved in signal transduction, oxidative stress defence, cell cycle regulation and heat shock responses was investigated with quantitative PCR. The data indicate that Euglena cells suffer stress upon short-term exposure to microgravity; various stress-induced genes were up-regulated. Of 32 tested genes, 18 were up-regulated, one down-regulated and the rest remained unaltered. These findings are in a good agreement with results from other research groups using other organisms. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

Gravitaxis of Euglena gracilis depends only partially on passive buoyancy

NASA Astrophysics Data System (ADS)

Richter, Peter R.; Schuster, Martin; Lebert, Michael; Streb, Christine; Häder, Donat-Peter

In darkness, the unicellular freshwater flagellate Euglena gracilis shows a pronounced negative gravitactic behavior, and the cells swim actively upward in the water column. Up to now it was unclear whether this behavior is based on a passive (physical) alignment mechanism (e.g., buoyancy due to a fore-aft asymmetry of the cell body) or on an active physiological mechanism. A sounding rocket experiment was performed in which the effect of sub-1g-accelerations (0.05, 0.08, 0.12, and 0.2g) on untreated living cells and immobilized (fixation with liquid nitrogen) cells was observed. By means of computerized image analysis the angles of the cells long axis with respect to the acceleration vector were analyzed in order to calculate and compare the reorientation kinetics of the immobilized cells versus that of the controls. In both groups, the reorientation kinetics depended on the dose, but the reorientation of the living cells was about five times faster than that of the immobilized cells. This indicates that in young cells gravitaxis can be explained by a physical mechanism only to a small extend. In older cultures, in which the cells often have a drop shaped cell body, the physical reorientation is considerably faster, and a more pronounced influence of passive alignment caused by fore/aft asymmetry (drag-gravity model) can not be excluded. In addition to these results, Euglena gracilis cells seem to respond very sensitively to small accelerations when they are applied after a longer microgravity period. The data indicate that gravitactic orientation occurred at an acceleration as low as 0.05g.

Amoebas and Euglenas and Paramecia...Oh, My!

ERIC Educational Resources Information Center

Farenga, Stephen; Joyce, Beverly

2000-01-01

Describes a partnership involving fifth grade students, preservice teachers, a classroom teacher, and a university faculty member in introducing students to the basic anatomy of a cell using three-dimensional models, analogies, and hands-on/minds-on activities. (ASK)

Microscopic Moves.

ERIC Educational Resources Information Center

Zurcher, Sandra L.

1991-01-01

Presents lessons to stimulate children's interest in science and develop communication skills by having students simulate the locomotion of an ameba, a paramecium, and an Euglena, and the cyclosis in the pondweed Elodea. Students practice communication skills by demonstrating their organism to the remainder of the class. (MDH)

Motion of Euglena gracilis: Active fluctuations and velocity distribution

NASA Astrophysics Data System (ADS)

Romanczuk, P.; Romensky, M.; Scholz, D.; Lobaskin, V.; Schimansky-Geier, L.

2015-07-01

We study the velocity distribution of unicellular swimming algae Euglena gracilis using optical microscopy and active Brownian particle theory. To characterize a peculiar feature of the experimentally observed distribution at small velocities we use the concept of active fluctuations, which was recently proposed for the description of stochastically self-propelled particles [Romanczuk, P. and Schimansky-Geier, L., Phys. Rev. Lett. 106, 230601 (2011)]. In this concept, the fluctuating forces arise due to internal random performance of the propulsive motor. The fluctuating forces are directed in parallel to the heading direction, in which the propulsion acts. In the theory, we introduce the active motion via the depot model [Schweitzer, et al., Phys. Rev. Lett. 80(23), 5044 (1998)]. We demonstrate that the theoretical predictions based on the depot model with active fluctuations are consistent with the experimentally observed velocity distributions. In addition to the model with additive active noise, we obtain theoretical results for a constant propulsion with multiplicative noise.

Removal of technetium from solution by algal flagellate Euglena gracilis.

PubMed

Ishii, Nobuyoshi; Uchida, Shigeo

2006-01-01

Based on limited data for the removal of radioactive (99)Tc by freshwater phytoplankton, it has been thought that phytoplankton are unsuitable for remediation of (99)Tc-contaminated waters. This work sought to verify the validity of this assumption by studying the removal of (99)TcO(4)(-) by freshwater and brackish water phytoplankton. The phytoplankton used were Euglena gracilis, Chlamydomonas pulsatilla, Chlorella vulgaris, and Spirulina platensis. Each of them was incubated for 63 d, and the removal of (99)Tc from solution was periodically determined. Significant removal of (99)Tc was observed only for E. gracilis, and the maximum removal was 70% of the total (99)Tc added. The killed cells of E. gracilis, however, removed hardly any (99)Tc. When E. gracilis cells were washed with fresh culture medium, only 13% of the total (99)Tc was desorbed. These results suggested that intracellular uptake of (99)Tc by E. gracilis occurred. These results are the first documented example of significant removal of (99)Tc by planktonic microalgae.

Sterling C. Robertson Dam and Limestone Lake on the Navasota River, Texas (Leon, Limestone and Robertson Counties).

DTIC Science & Technology

1976-04-01

Plicrocysti s sp. Synechococcus sp. Order Chamaesi phonal es Chamaesiphon sp. Order Hormogonal1es Anabaena sp. Anabaenopsis sp. Arthrospira sp...Sc-ytonerrn sp. Stig m rp. Spirulina sp. Kingdom Protista Phylum Euglenophyta (Euglenas) Order Euglenales Eugena sp. Phacus sp. Trache omonas sp

ß-1,3 glucan derived from Euglena gracilis and Algamune enhances innate immune responses of red drum (Sciaenops ocellatus L.)

USDA-ARS?s Scientific Manuscript database

To reduce susceptibility to stressors and diseases, immune-modulators such as ß-glucans have been proven effective tools to enhance the innate immune responses of fish. Consequently, commercial sources of this polysaccharide are becoming increasingly more available. AlgamuneTM is a commercial addi...

Compression and release dynamics of an active matter system of Euglena gracilis

NASA Astrophysics Data System (ADS)

Lam, Amy; Tsang, Alan C. H.; Ouellette, Nicholas; Riedel-Kruse, Ingmar

Active matter, defined as ensembles of self-propelled particles, encompasses a large variety of systems at all scales, from nanoparticles to bird flocks. Though various models and simulations have been created to describe the dynamics of these systems, experimental verification has been difficult to obtain. This is frequently due to the complex interaction rules which govern the particle behavior, in turn making systematic varying of parameters impossible. Here, we propose a model for predicting the system evolution of compression and release of an active system based on experiments and simulations. In particular, we consider ensembles of the unicellular, photo-responsive algae, Euglena gracilis, under light stimulation. By varying the spatiotemporal light patterns, we are able to finely adjust cell densities and achieve arbitrary non-homogeneous distributions, including compression into high-density aggregates of varying geometries. We observe the formation of depletion zones after the release of the confining stimulus and investigate the effects of the density distribution and particle rotational noise on the depletion. These results provide implications for defining state parameters which determine system evolution.

Solar Radiation Stress in Natural Acidophilic Biofilms of Euglena mutabilis Revealed by Metatranscriptomics and PAM Fluorometry.

PubMed

Puente-Sánchez, Fernando; Olsson, Sanna; Gómez-Rodriguez, Manuel; Souza-Egipsy, Virginia; Altamirano-Jeschke, Maria; Amils, Ricardo; Parro, Victor; Aguilera, Angeles

2016-02-01

The daily photosynthetic performance of a natural biofilm of the extreme acidophilic Euglena mutabilis from Río Tinto (SW, Spain) under full solar radiation was analyzed by means of pulse amplitude-modulated (PAM) fluorescence measurements and metatrascriptomic analysis. Natural E. mutabilis biofilms undergo large-scale transcriptomic reprogramming during midday due to a dynamic photoinhibition and solar radiation stress. Photoinhibition is due to UV radiation and not to light intensity, as revealed by PAM fluorometry analysis. In order to minimize the negative effects of solar radiation, our data supports the presence of a circadian rhythm in this euglenophyte that increases their opportunity to survive. Differential gene expression throughout the day (at 12:00, 20:00 and night) was monitored by massive Illumina parallel sequencing of metatranscriptomic libraries. The transcription pattern was altered in genes involved in Photosystem II stability and repair, UV damaged DNA repair, non-photochemical quenching and oxidative stress, supporting the photoinhibition detected by PAM fluorometry at midday. Copyright © 2016 Elsevier GmbH. All rights reserved.

Label-free chemical imaging of live Euglena gracilis by high-speed SRS spectral microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Wakisaka, Yoshifumi; Suzuki, Yuta; Tokunaga, Kyoya; Hirose, Misa; Domon, Ryota; Akaho, Rina; Kuroshima, Mai; Tsumura, Norimichi; Shimobaba, Tomoyoshi; Iwata, Osamu; Suzuki, Kengo; Nakashima, Ayaka; Goda, Keisuke; Ozeki, Yasuyuki

2016-03-01

Microbes, especially microalgae, have recently been of great interest for developing novel biofuels, drugs, and biomaterials. Imaging-based screening of live cells can provide high selectivity and is attractive for efficient bio-production from microalgae. Although conventional cellular screening techniques use cell labeling, labeling of microbes is still under development and can interfere with their cellular functions. Furthermore, since live microbes move and change their shapes rapidly, a high-speed imaging technique is required to suppress motion artifacts. Stimulated Raman scattering (SRS) microscopy allows for label-free and high-speed spectral imaging, which helps us visualize chemical components inside biological cells and tissues. Here we demonstrate high-speed SRS imaging, with temporal resolution of 0.14 seconds, of intracellular distributions of lipid, polysaccharide, and chlorophyll concentrations in rapidly moving Euglena gracilis, a unicellular phytoflagellate. Furthermore, we show that our method allows us to analyze the amount of chemical components inside each living cell. Our results indicate that SRS imaging may be applied to label-free screening of living microbes based on chemical information.

Light-stimulated Production of a Chloroplast-localized System for Protein Synthesis in Euglena gracilis1

PubMed Central

Reger, Bonnie J.; Smillie, R. M.; Fuller, R. C.

1972-01-01

Chloroplasts and proplastids isolated respectively from autotrophic and dark-adapted cells of Euglena gracilis strain Z incorporated 14C-l-leucine into protein. In each case the incorporation was inhibited by chloramphenicol (50% inhibition at about 5 μg/ml for chloroplasts and 30 μg/ml for proplastids), but not appreciably by cycloheximide at concentrations up to 200 μg/ml. Chloroplasts from autotrophic cells incorporated leucine into protein at rates of about 10 pg leucine per mg RNA in one minute, but isolated proplastids were only 5 to 10% as active. When dark-adapted cells were illuminated there was little increase in the activity of the chloroplast fraction during the first 12 hr. Between 12 and 24 hr, when there was a rapid increase in the rate of synthesis of chlorophyll, the capacity of the chloroplast fraction for protein synthesis increased markedly. Suppression of the formation of a chloroplast-localized system for protein synthesis by treating the cells with chloramphenicol and the lack of such an effect with cycloheximide suggests that certain of the proteins which form part of a functional chloroplast system for protein synthesis are themselves synthesized within the chloroplasts. PMID:16658126

Membrane skeletal proteins and their integral membrane protein anchors are targets for tyrosine and threonine kinases in Euglena.

PubMed

Fazio, M J; Da Silva, A C; Rosiere, T K; Bouck, G B

1995-01-01

Proteins of the membrane skeleton of Euglena gracilis were extensively phosphorylated in vivo and in vitro after incubation with [32P]-orthophosphate or gamma-[32P] ATP. Endogenous protein threonine/serine activity phosphorylated the major membrane skeletal proteins (articulins) and the putative integral membrane protein (IP39) anchor for articulins. The latter was also the major target for endogenous protein tyrosine kinase activity. A cytoplasmic domain of IP39 was specifically phosphorylated, and removal of this domain with papain eliminated the radiolabeled phosphoamino acids and eliminated or radically shifted the PI of the multiple isoforms of IP39. In gel kinase assays IP39 autophosphorylated and a 25 kDa protein which does not autophosphorylate was identified as a threonine/serine (casein) kinase. Plasma membranes from the membrane skeletal protein complex contained threonine/serine (casein) kinase activity, and cross-linking experiments suggested that IP39 was the likely source for this membrane activity. pH optima, cation requirements and heparin sensitivity of the detergent solubilized membrane activity were determined. Together these results suggest that protein kinases may be important modulators of protein assembly and function of the membrane skeleton of these protistan cells.

In situ proteo-metabolomics reveals metabolite secretion by the acid mine drainage bio-indicator, Euglena mutabilis

PubMed Central

Halter, David; Goulhen-Chollet, Florence; Gallien, Sébastien; Casiot, Corinne; Hamelin, Jérôme; Gilard, Françoise; Heintz, Dimitri; Schaeffer, Christine; Carapito, Christine; Van Dorsselaer, Alain; Tcherkez, Guillaume; Arsène-Ploetze, Florence; Bertin, Philippe N

2012-01-01

Euglena mutabilis is a photosynthetic protist found in acidic aquatic environments such as peat bogs, volcanic lakes and acid mine drainages (AMDs). Through its photosynthetic metabolism, this protist is supposed to have an important role in primary production in such oligotrophic ecosystems. Nevertheless, the exact contribution of E. mutabilis in organic matter synthesis remains unclear and no evidence of metabolite secretion by this protist has been established so far. Here we combined in situ proteo-metabolomic approaches to determine the nature of the metabolites accumulated by this protist or potentially secreted into an AMD. Our results revealed that the secreted metabolites are represented by a large number of amino acids, polyamine compounds, urea and some sugars but no fatty acids, suggesting a selective organic matter contribution in this ecosystem. Such a production may have a crucial impact on the bacterial community present on the study site, as it has been suggested previously that prokaryotes transport and recycle in situ most of the metabolites secreted by E. mutabilis. Consequently, this protist may have an indirect but important role in AMD ecosystems but also in other ecological niches often described as nitrogen-limited. PMID:22237547

β-1,3 glucan derived from Euglena gracilis and Algamune™ enhances innate immune responses of red drum (Sciaenops ocellatus L.).

PubMed

Yamamoto, Fernando Y; Yin, Fei; Rossi, Waldemar; Hume, Michael; Gatlin, Delbert M

2018-06-01

To reduce susceptibility to stressors and diseases, immune-modulators such as β-glucans have been proven effective tools to enhance the innate immune responses of fish. Consequently, commercial sources of this polysaccharide are becoming increasingly more available. Algamune™ is a commercial additive produced from Euglena gracilis, as a source of linear β-1,3-glucan. In order to evaluate the immunomodulatory effects of this β-glucan product, the present study assessed the innate immune parameters of red drum (Sciaenops ocellatus) exposed to Algamune™ ex vivo and in vivo. Isolated kidney phagocytes were incubated with graded concentrations (0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg L -1 ) of dried Euglena gracilis (Algamune™) as well as purified Paramylon (linear β-1,3 glucan). Increased bactericidal activity against Streptococcus iniae, and production of intracellular O 2 - anion superoxide were stimulated by both β-glucan sources. A reduced activity of extracellular anion superoxide was observed by the phagocytes incubated with Algamune ™. After corroborating the effectiveness of the glucan source ex vivo, a feeding trial was conducted using red drum juveniles (∼26.6 g initial weight). Fish were fed diets with graded levels of Algamune™ (0, 100, 200, 400 and 800 mg kg -1 ) twice daily for 21 days. No significant differences were detected regarding production performance parameters. At the end of the feeding trial, blood, intestinal content, and kidney were sampled. Intestinal microbiota from fecal material was analyzed through denaturing gradient gel electrophoresis (DGGE) and found to be similar among all treatments. No significant differences were detected for oxidative radical production from whole blood, and isolated phagocytes, and plasma lysozyme activity. However, the total hemolytic activity of red drum plasma was increased in fish fed 100 and 200 mg kg -1 of dietary Algamune™ when compared to fish fed the basal diet. Based on results from both ex vivo and in vivo trials, β-glucan from Algamune™ was demonstrated to have a moderate immunostimulatory effects on red drum. Copyright © 2018 Elsevier Ltd. All rights reserved.

Indications for acceleration-dependent changes of membrane potential in the flagellate Euglena gracilis.

PubMed

Richter, P R; Schuster, M; Meyer, I; Lebert, M; Häder, D-P

2006-12-01

The effects of the calcium sequester EGTA on gravitactic orientation and membrane potential changes in the unicellular flagellate Euglena gracilis were investigated during a recent parabolic-flight experiment aboard of an Airbus A300. In the course of a flight parabola, an acceleration profile is achieved which yields subsequently about 20 s of hypergravity (1.8 g(n)), about 20 s of microgravity, and another 20 s of hypergravity phases. The movement behavior of the cells was investigated with real-time, computer-based image analysis. Membrane potential changes were detected with a newly developed photometer which measures absorption changes of the membrane potential-sensitive probe oxonol VI. To test whether the data obtained by the oxonol device were reliable, the signal of non-oxonol-labelled cells was recorded. In these samples, no absorption shift was detected. Changes of the oxonol VI signals indicate that the cells depolarize during acceleration (very obvious in the step from microgravity to hypergravity) and slightly hyperpolarize in microgravity, which can possibly be explained with the action of Ca-ATPases. These signals (mainly the depolarization) were significantly suppressed in the presence of EGTA (5 mM). Gravitaxis in parallel was also inhibited after addition of EGTA. Initially, negative gravitaxis was inverted into a positive one. Later, gravitaxis was almost undetectable.

Analysis and Characterization of 3-(3,4-Dichlorophenyl)-1,1-Dimethylurea (DCMU)-resistant Euglena

PubMed Central

Calvayrac, Régis; Bomsel, Jean-Loup; Laval-Martin, Danielle

1979-01-01

Cultures of Euglena gracilis Klebs strain Z Pringsheim were grown photoorganotrophically in the presence of different concentrations of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) in the range of 0.05 to 250 micromolar. Cultures were serially transferred and various metabolic parameters were followed for 10 weeks. A process of adaptation occurred which was divided operationally into three phases. A phase of ultrastructural disorganization occurred, succeeded by a recovery phase; their intensity and duration were functions of the dose of DCMU. A stable adaptation phase then ensued. This phase was observed in all cultures except that exposed to the highest DCMU concentration. Adapted cells from all of the DCMU cultures contained twice the protein and half the paramylon of the control cells and thus utilized the carbon source to accumulate cellular reserves with only half the efficiency of controls. DCMU affected cellular metabolism as well as photosynthesis. The energy charge remained at high levels throughout adaptation, although the size of the adenylate pool was half that of controls at the disorganized phase. At this stage the ultrastructure of chloroplasts and mitochondria was considerably modified. The progressive changes of the parameters studied appeared to affect all of the cells in a given culture. Images PMID:16660827

Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting

PubMed Central

Yamada, Koji; Suzuki, Hideyuki; Takeuchi, Takuto; Kazama, Yusuke; Mitra, Sharbanee; Abe, Tomoko; Goda, Keisuke; Suzuki, Kengo; Iwata, Osamu

2016-01-01

Euglena gracilis, a microalgal species of unicellular flagellate protists, has attracted much attention in both the industrial and academic sectors due to recent advances in the mass cultivation of E. gracilis that have enabled the cost-effective production of nutritional food and cosmetic commodities. In addition, it is known to produce paramylon (β-1,3-glucan in a crystalline form) as reserve polysaccharide and convert it to wax ester in hypoxic and anaerobic conditions–a promising feedstock for biodiesel and aviation biofuel. However, there remain a number of technical challenges to be solved before it can be deployed in the competitive fuel market. Here we present a method for efficient selective breeding of live oil-rich E. gracilis with fluorescence-activated cell sorting (FACS). Specifically, the selective breeding method is a repetitive procedure for one-week heterotrophic cultivation, staining intracellular lipids with BODIPY505/515, and FACS-based isolation of top 0.5% lipid-rich E. gracilis cells with high viability, after inducing mutation with Fe-ion irradiation to the wild type (WT). Consequently, we acquire a live, stable, lipid-rich E. gracilis mutant strain, named B1ZFeL, with 40% more lipid content on average than the WT. Our method paves the way for rapid, cost-effective, energy-efficient production of biofuel. PMID:27212384

A step towards on-chip biochemical energy cascade of microorganisms: carbon dioxide generation induced by ethanol fermentation in 3D printed modular lab-on-a-chip

NASA Astrophysics Data System (ADS)

Podwin, A.; Kubicki, W.; Adamski, K.; Walczak, R.; Dziuban, J. A.

2016-11-01

The concept of biochemical energy cascade of microorganisms towards oxygen generation in 3D printed lab-on-a-chip has been presented. In this work, carbon dioxide - a product of ethanol fermentation of yeasts has been utilized to enable light-initialized photosynthesis of euglenas and as a result of their metabolic transitions produce pure oxygen.

Advantages of Environmental Scanning Electron Microscopy in Studies of Microorganisms

DTIC Science & Technology

1993-01-01

external structures, cultures of Aspergillus sp. and Cunninghamella sp. but details of cytoplasmic organization were lacking were maintained on corn meal ... agar . (Fig. 7). Uncoated, freeze-dried Euglena sp. cells exam- ined with either ESD or BSE detectors did not exhibit Specimen Preparation for ESEM...approximately Agar plugs from sporulating cultures of Cunning- 1.0 pLm in diameter (Fig. 23). Many of the vesicles ap- hamella sp., Mucor sp., and Aspergillus

High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy

NASA Astrophysics Data System (ADS)

Guo, Baoshan; Lei, Cheng; Ito, Takuro; Yaxiaer, Yalikun; Kobayashi, Hirofumi; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

2017-02-01

The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, microalgal biofuel is expected to play a key role in reducing the detrimental effects of global warming since microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid contents and fail to characterize a diverse population of microalgal cells with single-cell resolution in a noninvasive and interference-free manner. Here we demonstrate high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy. In particular, we use Euglena gracilis - an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement) within lipid droplets. Our optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch phase-contrast microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase contents of every single cell at a high throughput of 10,000 cells/s. We characterize heterogeneous populations of E. gracilis cells under two different culture conditions to evaluate their lipid production efficiency. Our method holds promise as an effective analytical tool for microalgaebased biofuel production.

Generalized receptor law governs phototaxis in the phytoplankton Euglena gracilis

PubMed Central

Giometto, Andrea; Altermatt, Florian; Maritan, Amos; Stocker, Roman; Rinaldo, Andrea

2015-01-01

Phototaxis, the process through which motile organisms direct their swimming toward or away from light, is implicated in key ecological phenomena (including algal blooms and diel vertical migration) that shape the distribution, diversity, and productivity of phytoplankton and thus energy transfer to higher trophic levels in aquatic ecosystems. Phototaxis also finds important applications in biofuel reactors and microbiopropellers and is argued to serve as a benchmark for the study of biological invasions in heterogeneous environments owing to the ease of generating stochastic light fields. Despite its ecological and technological relevance, an experimentally tested, general theoretical model of phototaxis seems unavailable to date. Here, we present accurate measurements of the behavior of the alga Euglena gracilis when exposed to controlled light fields. Analysis of E. gracilis’ phototactic accumulation dynamics over a broad range of light intensities proves that the classic Keller–Segel mathematical framework for taxis provides an accurate description of both positive and negative phototaxis only when phototactic sensitivity is modeled by a generalized “receptor law,” a specific nonlinear response function to light intensity that drives algae toward beneficial light conditions and away from harmful ones. The proposed phototactic model captures the temporal dynamics of both cells’ accumulation toward light sources and their dispersion upon light cessation. The model could thus be of use in integrating models of vertical phytoplankton migrations in marine and freshwater ecosystems, and in the design of bioreactors. PMID:25964338

Conducting a battery of bioassays for gold nanoparticles to derive guideline value for the protection of aquatic ecosystems.

PubMed

Nam, Sun-Hwa; Shin, Yu-Jin; Lee, Woo-Mi; Kim, Shin Woong; Kwak, Jin Il; Yoon, Sung-Ji; An, Youn-Joo

2015-05-01

Gold nanoparticles (Au-NPs) are used in many applications, including the manufacture of products like cosmetics, paints, and electrochemical immunosensors, and in the detection, diagnosis, and treatment of tumors. However, there are no legal or recommended guidelines for protecting aquatic ecosystems from Au-NPs. In this study, we conducted a battery of bioassays and present toxicity values for two bacteria, one alga, one euglena, three cladoceran, and two fish species that were exposed to Au-NPs. Guideline values for protecting aquatic ecosystems from Au-NPs were derived using methods that are generally used to derive water-quality guidelines and are used in Australia, New Zealand, Canada, the European Community (EC), and the USA. Au-NPs had adverse effects on all test species, including growth inhibition of both bacteria, the alga, and the euglena; mortality and immobilization in the three cladocerans; and developmental malformations in the embryos and larvae of the two fish. Guideline values of 0.15 and 0.04 × 10(10) particles/mL were derived for Au-NPs using a species sensitivity distribution (SSD) and assessment factor. The guideline value derived for Au-NPs using an assessment factor was more stringent than that derived using SSD. This is the first study to derive guideline values for nanoparticles in water environments.

Generalized receptor law governs phototaxis in the phytoplankton Euglena gracilis.

PubMed

Giometto, Andrea; Altermatt, Florian; Maritan, Amos; Stocker, Roman; Rinaldo, Andrea

2015-06-02

Phototaxis, the process through which motile organisms direct their swimming toward or away from light, is implicated in key ecological phenomena (including algal blooms and diel vertical migration) that shape the distribution, diversity, and productivity of phytoplankton and thus energy transfer to higher trophic levels in aquatic ecosystems. Phototaxis also finds important applications in biofuel reactors and microbiopropellers and is argued to serve as a benchmark for the study of biological invasions in heterogeneous environments owing to the ease of generating stochastic light fields. Despite its ecological and technological relevance, an experimentally tested, general theoretical model of phototaxis seems unavailable to date. Here, we present accurate measurements of the behavior of the alga Euglena gracilis when exposed to controlled light fields. Analysis of E. gracilis' phototactic accumulation dynamics over a broad range of light intensities proves that the classic Keller-Segel mathematical framework for taxis provides an accurate description of both positive and negative phototaxis only when phototactic sensitivity is modeled by a generalized "receptor law," a specific nonlinear response function to light intensity that drives algae toward beneficial light conditions and away from harmful ones. The proposed phototactic model captures the temporal dynamics of both cells' accumulation toward light sources and their dispersion upon light cessation. The model could thus be of use in integrating models of vertical phytoplankton migrations in marine and freshwater ecosystems, and in the design of bioreactors.

Molecular characterization of phytoplankton dissolved organic matter (DOM) and sulfur components using high resolution Orbitrap mass spectrometry.

PubMed

Mangal, Vaughn; Stock, Naomi L; Guéguen, Celine

2016-03-01

Orbitrap high resolution mass spectrometry (HRMS) with electrospray ionization in both positive and negative polarity was conducted on Suwannee River fulvic acid (SRFA), Pony Lake fulvic acid (PLFA) standards, and dissolved organic matter (DOM) released by freshwater phytoplankton (Scenedesmus obliquus, Euglena mutabilis, and Euglena gracilis). Three-dimensional van Krevelen diagrams expressing various oxygenation states of sulfur molecules and abundance plots of sulfur-containing species were constructed. Orbitrap HRMS analysis of SRFA found a high density of peaks in the lignin region (77 %) and low density of protein material (6.53 %), whereas for PLFA, 25 % of the total peaks were lignin related compared to 56 % of peaks in protein regions, comparable with other HRMS studies. Phytoplankton-derived DOM of S. obliquus, E. mutabilis, and E. gracilis was dominated by protein molecules at respective percentages of 36, 46, and 49 %, and is consistent with previous experiments examining phytoplankton-derived DOM composition. The normalized percentage of SO-containing compounds was determined among the three phytoplankton to be 56 % for Scenedesmus, 54 % for E. mutabilis, and 47 % for E. gracilis, suggesting variation between sulfur content in phytoplankton-derived DOM and differences in metal binding capacities. These results suggest the level of resolution by Orbitrap mass spectrometry is sufficient for preliminary characterization of phytoplankton DOM at an affordable cost relative to other HRMS techniques.

Effects of near ultraviolet and green radiations on plant growth

DOE Office of Scientific and Technical Information (OSTI.GOV)

Klein, R.M.; Edsall, P.C.; Gentile, A.C.

Selective removal of near ultraviolet and green wavelengths from white light permitted enhanced growth of marigold, tomato, corn, and Impatiens plants, Chlamydomonas cells and the mycelium of Sordaria. Additions of near ultraviolet and green radiations caused repressions in the growth of marigold and Sordaria. These wavelengths do not alter the oxidative mechanisms of mitochondria, intact algal cells or marigold leaf tissues. The capacity for chlorophyll and carotenoid synthesis by Euglena cells was unaffected by these wavelengths. 23 references, 2 figures, 4 tables.

The role of algae in mine drainage bioremediation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Davison, J.

1990-06-01

The effect of mine drainage effluent on aquatic ecosystems has been abundantly documented and remediation efforts to data have always been costly and temporary at best. Bioremediation, using natural environmental microbes, to treat acid mine drainage has shown great promise as an affordable, permanent treatment. At Lambda, we used mixatrophic cultures of bacteria, algae, protozoans and fungal groups on four different jobs and it has proven effective. The role of two particular algal groups, the Euglena mutabilis and the Ochramonas sp. are particularly of phycological interest.

Genetic control of enzyme formation. Final technical report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mills, S. E.

1978-07-26

Research progress is reported on work on tryptophan biosynthesis in Euglena gracilis and higher plants. The experimental data provide an outline of the general evolution of the pathway. Structural analyses of the pathway proteins by quantitative immunochemical methods have been completed; this was done with the anthranilate synthase-1 phosphoribosyl transferase complex in Escherichia coli. An examination of the evolution, in the Enterobacteriaceae, of the enzyme activities anthranilate synthase and anthranilate-5-1 phosphoribosyl-1-pyrophosphate phosphoribosyltransferase has been begun. (ACR)

In vitro degradation of ribosomes.

PubMed

Mora, G; Rivas, A

1976-12-01

The cytoplasmic ribosomes from Euglena gracilis var. bacillaris are found to be of two types taking into consideration their stability "in vitro". In the group of unstable ribosomes the large subunit is degraded. The other group apparently does not suffer any degradation under the conditions described. However the RNAs extracted from both types of ribosomes are degraded during sucrose density gradients. The degradation of the largest RNA species has been reported previously, but no comment has been made about the stability of the ribosome itself.

Air pollutant production by algal cell cultures

NASA Technical Reports Server (NTRS)

Fong, F.; Funkhouser, E. A.

1982-01-01

The production of phytotoxic air pollutants by cultures of Chlorella vulgaris and Euglena gracilis is considered. Algal and plant culture systems, a fumigation system, and ethylene, ethane, cyanide, and nitrogen oxides assays are discussed. Bean, tobacco, mustard green, cantaloupe and wheat plants all showed injury when fumigated with algal gases for 4 hours. Only coleus plants showed any resistance to the gases. It is found that a closed or recycled air effluent system does not produce plant injury from algal air pollutants.

Euglena gracilis and Trypanosomatids possess common patterns in predicted mitochondrial targeting presequences.

PubMed

Krnáčová, Katarína; Vesteg, Matej; Hampl, Vladimír; Vlček, Čestmír; Horváth, Anton

2012-10-01

Euglena gracilis possessing chloroplasts of secondary green algal origin and parasitic trypanosomatids Trypanosoma brucei, Trypanosoma cruzi and Leishmania major belong to the protist phylum Euglenozoa. Euglenozoa might be among the earliest eukaryotic branches bearing ancestral traits reminiscent of the last eukaryotic common ancestor (LECA) or missing features present in other eukaryotes. LECA most likely possessed mitochondria of endosymbiotic α-proteobacterial origin. In this study, we searched for the presence of homologs of mitochondria-targeted proteins from other organisms in the currently available EST dataset of E. gracilis. The common motifs in predicted N-terminal presequences and corresponding homologs from T. brucei, T. cruzi and L. major (if found) were analyzed. Other trypanosomatid mitochondrial protein precursor (e.g., those involved in RNA editing) were also included in the analysis. Mitochondrial presequences of E. gracilis and these trypanosomatids seem to be highly variable in sequence length (5-118 aa), but apparently share statistically significant similarities. In most cases, the common (M/L)RR motif is present at the N-terminus and it is probably responsible for recognition via import apparatus of mitochondrial outer membrane. Interestingly, this motif is present inside the predicted presequence region in some cases. In most presequences, this motif is followed by a hydrophobic region rich in alanine, leucine, and valine. In conclusion, either RR motif or arginine-rich region within hydrophobic aa-s present at the N-terminus of a preprotein can be sufficient signals for mitochondrial import irrespective of presequence length in Euglenozoa.

Growth promotion of Euglena gracilis by ferulic acid from rice bran.

PubMed

Zhu, Jiangyu; Wakisaka, Minato

2018-02-08

A significant growth promotion of Euglena gracilis was achieved by simply adding ferulic acid from rice bran without diminishing the accumulation of valuable products like paramylon. E. gracilis is a freshwater microalga that is widely applied in cosmetics, food, medicine, and supplements, and it is considered a potential source of biofuel. It is therefore important to enhance its yield at a lower cost for its commercial viability. Introducing a growth regulator derived from agro waste is considered a cheaper and safer strategy to improve biomass productivity compared with other alternatives such as implementing genetic engineering or adding nutrients and plant hormones as growth stimulator. The effect of ferulic acid derived from rice bran on the growth and metabolism of E. gracilis was investigated in this study. To aid in the dissolution of ferulic acid, 1% dimethyl sulfoxide (DMSO) was added to Cramer-Myers medium. Ferulic acid could alleviate the inhibitory effect of DMSO and significantly promoted the growth of E. gracilis. It was found that cell density was 2.5 times greater than that of the control group and 3.6 times greater than that of the negative control group when 500 mg/L of ferulic acid was added. In addition, the photosynthetic pigment content, especially chlorophyll a, increased with increasing ferulic acid concentrations. The total paramylon production would also be enhanced by ferulic acid since the number of cells increased without reducing the cellular content of paramylon.

High-speed stimulated Raman scattering microscopy for studying the metabolic diversity of motile Euglena gracilis

NASA Astrophysics Data System (ADS)

Suzuki, Y.; Wakisaka, Y.; Iwata, O.; Nakashima, A.; Ito, T.; Hirose, M.; Domon, R.; Sugawara, M.; Tsumura, N.; Watarai, H.; Shimobaba, T.; Suzuki, K.; Goda, K.; Ozeki, Y.

2017-02-01

Microalgae have been receiving great attention for their ability to produce biomaterials that are applicable for food supplements, drugs, biodegradable plastics, and biofuels. Among such microalgae, Euglena gracilis has become a popular species by virtue of its capability of accumulating useful metabolites including paramylon and lipids. In order to maximize the production of desired metabolites, it is essential to find ideal culturing conditions and to develop efficient methods for genetic transformation. To achieve this, understanding and controlling cell-to-cell variations in response to external stress is essential, with chemically specific analysis of microalgal cells including E. gracilis. However, conventional analytical tools such as fluorescence microscopy and spontaneous Raman scattering are not suitable for evaluation of diverse populations of motile microalgae, being restricted either by the requirement for fluorescent labels or a limited imaging speed, respectively. Here we demonstrate video-rate label-free metabolite imaging of live E. gracilis using stimulated Raman scattering (SRS) - an optical spectroscopic method for probing the vibrational signatures of molecules with orders of magnitude higher sensitivity than spontaneous Raman scattering. Our SRS's highspeed image acquisition (27 metabolite images per second) allows for population analysis of live E. gracilis cells cultured under nitrogen-deficiency - a technique for promoting the accumulation of paramylon and lipids within the cell body. Thus, our SRS system's fast imaging capability enables quantification and analysis of previously unresolvable cell-to-cell variations in the metabolite accumulation of large motile E. gracilis cell populations.

Composition, quaternary structure, and catalytic properties of D-ribulose-1, 5-bisphosphate carboxylase from Euglena gracilis.

PubMed

McFadden, B A; Lord, J M; Rowe, A; Dilks, S

1975-05-01

D-Ribulose-1,5-bisphosphate carboxylase has been purified in one step by sedimenting extracts of autotrophically-grown Euglena gracilis into a linear 0.2-0.8 M sucrose density gradient. The resultant product was pure by the criteria of disc electrophoresis in gels polymerized from 5 or 7.5% acrylamide and sedimentation. The molecular weight of the enzyme estimated by density gradient centrifugation and electrophoresis in gels polymerized from various concentrations of acrylamide was 5.25 X 10(5). The S20,W was 16.4 S. Dissociation and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate established that the enzyme was composed of two types of subunits (mr 50,000 and 15,000). The oligomeric structure was visualized through negative staining and transmission electron microscopy leading to a model for the quaternary structure. Although the enzyme was moderately unstable, the estimated maximal specific activity was 1.6 mumol CO2 fixed min-1 mg protien-1 at 30 degrees C and pH 8.0 Km values were 2.2 m M, 15. 1 MUM and 0.63 mM for Mg2+, ribulose 1,5-bisphosphate, and CO2, respectively, when measured under air. 6-Phospho-D-gluconate was a noncompetitive inhibitor with respect to ribulose 1,5-bisphosphate (Ki = 0.04 mM). Oxygen was a competitive inhibitor with respect to CO2 suggesting that the enzyme was also an oxygenase. The latter was confirmed by experiments showing a molar equivalence between ribulose-1,5-bisphosphate-dependent oxygen consumption and phosphoglycerate production.

Toxicity assessment of a common laundry detergent using the freshwater flagellate Euglena gracilis.

PubMed

Azizullah, Azizullah; Richter, Peter; Häder, Donat-Peter

2011-09-01

Synthetic detergents are among the commonly used chemicals in everyday life. Detergents, reaching aquatic environments through domestic and municipal wastewater, can cause many different effects in aquatic organisms. The present study was aimed at the toxicity evaluation of a commonly used laundry detergent, Ariel, using the freshwater flagellate Euglena gracilis as a biotest organism. Different parameters of the flagellate like motility, swimming velocity, cell shape, gravitactic orientation, photosynthesis and concentration of light harvesting pigments were used as end points for the toxicity assessment. No Observed Effect Concentration (NOEC) and EC(50) values were calculated for the end point parameters at four different incubation times, i.e. 0, 6, 24 and 72 h. After 72 h incubation, swimming velocity of the cells was found to be the most sensitive parameter giving NOEC and EC(50) values of 10.8 and 34 mg L(-1), respectively. After 72 h exposure to the detergent, chlorophyll a and total carotenoids were significantly decreased in cultures treated with Ariel at concentrations of 50 mg L(-1) and above while chlorophyll b significantly decreased at concentrations above 750 mg L(-1). The maximum inhibitory effect on the quantum yield of photosystem II was observed after 24 h exposure and thereafter a recovery trend was observed. Motility, gravitaxis and cell shape were strongly impaired immediately upon exposure to the detergent, but with increasing exposure time these parameters showed acclimatization to the stress and thus the NOEC values obtained after 72 h were higher than those immediately after exposure. Copyright © 2011 Elsevier Ltd. All rights reserved.

Improvement of kinetics, yield, and colloidal stability of biogenic gold nanoparticles using living cells of Euglena gracilis microalga

NASA Astrophysics Data System (ADS)

Dahoumane, Si Amar; Yéprémian, Claude; Djédiat, Chakib; Couté, Alain; Fiévet, Fernand; Coradin, Thibaud; Brayner, Roberta

2016-03-01

Recent years have witnessed a boom in the biosynthesis of a large variety of nanomaterials using different biological resources among which algae-based entities have been gaining much more attention within the community of material scientists worldwide. In our previously published findings, we explored some factors that governed the biofabrication of gold nanoparticles using living cultures of microalgae, such as the utilized microalgal genera, the phylum they belong to, and the impact of tetrachloroauric acid concentrations on the ability of these strains to perform the biosynthesis of gold nanoparticles once in contact with these cations. As a follow-up, we present in this paper an improvement of the features of bioproduced gold colloids using living cells of Euglena gracilis microalga when this species is grown under either mixotrophic or autotrophic conditions, i.e., exposed to light and grown in an organic carbon-enriched culture medium versus under autotrophic conditions. As an outcome to this alteration, the growth rate of this photosynthetic microorganism is multiplied 7-8 times when grown under mixotrophic conditions compared to autotrophic ones. Therefore, the yield, the kinetics, and the colloidal stability of the biosynthesized gold nanoparticles are dramatically enhanced. Moreover, the shape and the size of the as-produced nano-objects via this biological method are affected. In addition to round-shaped gold nanoparticles, particular shapes, such as triangles and hexagons, appear. These findings add up to the amassed knowledge toward the design of photobioreactors for the scalable and sustainable production of interesting nanomaterials.

Operation of an enclosed aquatic ecosystem in the Shenzhou-8 mission

NASA Astrophysics Data System (ADS)

Li, Xiaoyan; Richter, Peter R.; Hao, Zongjie; An, Yanjun; Wang, Gaohong; Li, Dunhai; Liu, Yongding; Strauch, Sebastian M.; Schuster, Martin; Haag, Ferdinand W.; Lebert, Michael

2017-05-01

Long- term spaceflight needs reliable Biological life support systems (BLSS) to supply astronauts with enough food, fresh air and recycle wasters, but the knowledge about the operation pattern and controlling strategy is rear. For this purpose, a miniaturized enclosed aquatic ecosystem was developed and flown on the Chinese spaceship Shenzhou-8. The system with a total volume of about 60 mL was separated into two chambers by means of a gas transparent membrane. The lower chamber was inoculated with Euglena gracilis cells, and the upper chamber was cultured with Chlorella cells and three snails. After 17.5 days flight, the samples were analyzed. It was found that all snails in the ground module (GM) were alive, while in the flight module (FM) only one snail survived. The total cell numbers, assimilation of nutrients like nitrogen and phosphorus, soluble proteins and carbohydrate contents showed a decrease in FM than in GM. The correlation analysis showed upper chambers of both FM and GM had the same positive and negative correlation factors, while differential correlation was found in lower chambers. These results suggested primary productivity in the enclosed system decreased in microgravity, accompanied with nutrients assimilation. The FM chamber endured lacking of domination species to sustain the system development and GM chamber endured richness in population abundance. These results implied photosynthesis intensity should be reduced to keep the system healthy. More Chlorella but less Euglena might be a useful strategy to sustain system stability. It is the first systematic analysis of enclosed systems in microgravity.

Unusual features of fibrillarin cDNA and gene structure in Euglena gracilis: evolutionary conservation of core proteins and structural predictions for methylation-guide box C/D snoRNPs throughout the domain Eucarya.

PubMed

Russell, Anthony G; Watanabe, Yoh-ichi; Charette, J Michael; Gray, Michael W

2005-01-01

Box C/D ribonucleoprotein (RNP) particles mediate O2'-methylation of rRNA and other cellular RNA species. In higher eukaryotic taxa, these RNPs are more complex than their archaeal counterparts, containing four core protein components (Snu13p, Nop56p, Nop58p and fibrillarin) compared with three in Archaea. This increase in complexity raises questions about the evolutionary emergence of the eukaryote-specific proteins and structural conservation in these RNPs throughout the eukaryotic domain. In protists, the primarily unicellular organisms comprising the bulk of eukaryotic diversity, the protein composition of box C/D RNPs has not yet been extensively explored. This study describes the complete gene, cDNA and protein sequences of the fibrillarin homolog from the protozoon Euglena gracilis, the first such information to be obtained for a nucleolus-localized protein in this organism. The E.gracilis fibrillarin gene contains a mixture of intron types exhibiting markedly different sizes. In contrast to most other E.gracilis mRNAs characterized to date, the fibrillarin mRNA lacks a spliced leader (SL) sequence. The predicted fibrillarin protein sequence itself is unusual in that it contains a glycine-lysine (GK)-rich domain at its N-terminus rather than the glycine-arginine-rich (GAR) domain found in most other eukaryotic fibrillarins. In an evolutionarily diverse collection of protists that includes E.gracilis, we have also identified putative homologs of the other core protein components of box C/D RNPs, thereby providing evidence that the protein composition seen in the higher eukaryotic complexes was established very early in eukaryotic cell evolution.

Temporal change of photophobic step-up responses of Euglena gracilis investigated through motion analysis

PubMed Central

Ozasa, Kazunari; Won, June; Song, Simon; Tamaki, Shun; Ishikawa, Takahiro; Maeda, Mizuo

2017-01-01

The adaptation to a strong light is one of the essential characteristics of green algae, yet lacking relatively the information about the photophobic responses of Eukaryotic microalgae. We investigated the photophobic step-up responses of Euglena gracilis over a time course of several hours with alternated repetition of blue-light pulse illumination and spatially patterned blue-light illumination. Four distinctive photophobic motions in response to strong blue light were identified in a trace image analysis, namely on-site rotation, running and tumbling, continuous circular swimming, and unaffected straightforward swimming. The cells cultured in autotrophic conditions under weak light showed mainly the on-site rotation response at the beginning of blue-light illumination, but they acquired more blue-light tolerant responses of running and tumbling, circular swimming, or straightforward swimming. The efficiency of escaping from a blue-light illuminated area improved markedly with the development of these photophobic motions. Time constant of 3.0 h was deduced for the evolution of photophobic responses of E. gracilis. The nutrient-rich metabolic status of the cells resulting from photosynthesis during the experiments, i.e., the accumulation of photosynthesized nutrient products in balance between formation and consumption, was the main factor responsible for the development of photophobic responses. The reduction-oxidation status in and around E. gracilis cells did not affect their photophobic responses significantly, unlike the case of photophobic responses and phototaxis of Chlamydomonas reinhardtii cells. This study shows that the evolution of photophobic motion type of E. gracilis is dominated mainly by the nutrient metabolic status of the cells. The fact suggests that the nutrient-rich cells have a higher threshold for switching the flagellar motion from straightforward swimming to rotation under a strong light. PMID:28234984

High-Throughput Accurate Single-Cell Screening of Euglena gracilis with Fluorescence-Assisted Optofluidic Time-Stretch Microscopy.

PubMed

Guo, Baoshan; Lei, Cheng; Ito, Takuro; Jiang, Yiyue; Ozeki, Yasuyuki; Goda, Keisuke

2016-01-01

The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, algal biofuel is expected to play a key role in alleviating global warming since algae absorb atmospheric CO2 via photosynthesis. Among various algae for fuel production, Euglena gracilis is an attractive microalgal species as it is known to produce wax ester (good for biodiesel and aviation fuel) within lipid droplets. To date, while there exist many techniques for inducing microalgal cells to produce and accumulate lipid with high efficiency, few analytical methods are available for characterizing a population of such lipid-accumulated microalgae including E. gracilis with high throughout, high accuracy, and single-cell resolution simultaneously. Here we demonstrate high-throughput, high-accuracy, single-cell screening of E. gracilis with fluorescence-assisted optofluidic time-stretch microscopy-a method that combines the strengths of microfluidic cell focusing, optical time-stretch microscopy, and fluorescence detection used in conventional flow cytometry. Specifically, our fluorescence-assisted optofluidic time-stretch microscope consists of an optical time-stretch microscope and a fluorescence analyzer on top of a hydrodynamically focusing microfluidic device and can detect fluorescence from every E. gracilis cell in a population and simultaneously obtain its image with a high throughput of 10,000 cells/s. With the multi-dimensional information acquired by the system, we classify nitrogen-sufficient (ordinary) and nitrogen-deficient (lipid-accumulated) E. gracilis cells with a low false positive rate of 1.0%. This method holds promise for evaluating cultivation techniques and selective breeding for microalgae-based biofuel production.

A new approach for biological online testing of stack gas condensate from municipal waste incinerators.

PubMed

Elsner, Dorothea; Fomin, Anette

2002-01-01

A biological testing system for the monitoring of stack gas condensates of municipal waste incinerators has been developed using Euglena gracilis as a test organism. The motility, velocity and cellular form of the organisms were the endpoints, calculated by an image analysis system. All endpoints showed statistically significant changes in a short time when organisms were exposed to samples collected during combustion situations with increased pollutant concentrations. The velocity of the organisms proved to be the most appropriate endpoint. A semi-continuous system with E. gracilis for monitoring stack gas condensate is proposed, which could result in an online system for testing stack gas condensates in the future.

Choline and Inositol Distribution in Algae and Fungi1

PubMed Central

Ikawa, Miyoshi; Borowski, Paul T.; Chakravarti, Ashima

1968-01-01

Inositol and choline were present in varying amounts among the species of Rhodophyta, Phaeophyta, Chlorophyta, and Euglenophyta examined. However, in the two members of the order Fucales (division Phaeophyta) examined, no detectable amounts of choline were found. In contrast, the species of Cyanophyta examined contained no detectable amounts of either choline or inositol. All species of the fungal classes Phycomyceteae, Ascomyceteae, and Basidiomyceteae collected contained both inositol and choline in varying amounts. The red, brown, and blue-green algae usually contained much less inositol and choline than do plant and animals sources, but the fungi and the algae Chlorella and Euglena contained amounts comparable to those present in plant sources. PMID:5647522

The evolution of blue-greens and the origins of chloroplasts

NASA Technical Reports Server (NTRS)

Schwartz, R. M.; Dayhoff, M. O.

1981-01-01

All of the available molecular data support the theory that the chloroplasts of eukaryote cells were originally free-living blue-greens. Of great interest is what the relationships are between contemporary types of blue-greens and eukaryote chloroplasts and whether the chloroplasts of the various eukaryotes are the result of one or more than one symbiosis. By combining information from phylogenetic trees based on cytochrome c6 and 2Fe-2S ferredoxin sequences, it is shown that the chloroplasts of a number of eukaryote algae as well as the protist Euglena are polyphyletic; the chloroplasts of green algae and the higher plants may be the result of a single symbiosis.

Lipid Biomarkers and Carbon Isotope Ratios of Lipids Isolated from Acid Mine Drainage Biofilms: Dual Biosignatures for Eukaryotic Evolution and Oxygenation of Primitive Earth

NASA Astrophysics Data System (ADS)

Dasgupta, S.; Fang, J.; Zhang, L.; Li, J.

2012-12-01

Lipid analysis and carbon isotope ratios (δ13C) of lipids in biofilms in an acid mine drainage site (AMD) site in western Indiana revealed unique biogeochemical signatures of microeukaryotes, never recorded before. Dominance of photosynthetic microeukaryote Euglena was indicated by the detection of abundant phytadiene, phytol, phytanol, polyunsaturated n-alkenes, polyunsaturated fatty acids, short-chain (C25-32) wax esters (WE), ergosterol, and tocopherols. The WE were probably synthesized in mitochondria under anoxic conditions by the reverse β-oxidation pathway, whereas the sterols (ergosterol and ergosta-7,22-dien-3β-ol) were likely synthesized in the cytosol in the presence of molecular oxygen. The dual aerobic and anaerobic biosynthetic pathways of Euglena may be a response to survive the recurring anoxic and oxic conditions in primitive Earth, whereby microeukaryotes retained this mechanism of conserved compartmentalization within their physiology to evolve and diversify in extreme conditions. Hydrocarbons, including n-alkenes, phytadienes, and wax esters showed heavy δ13C values than usual. The primary cause for the 13C-enrichment can be attributed to a CO2-limiting system that exists in the AMD, which is further regulated by the pH of the AMD. Floating biofilms BF2, 4, and 6 showed more depleted δ13C values for phytadienes and n-alkenes (average of -23.6‰) as compared to benthic biofilm BF5 (average of -20.8‰), indicating that physiology plays an important role in isotopic discrimination. 13C-enriched values of the esters could result from kinetic isotope effects at two branch points (pyruvate and/or acetyl CoA) in the biosynthetic pathway. Our understanding of biogeochemical conditions in this AMD environment would allow us to identify unique sets of biosignatures that can act as a proxy in deciphering the links between eukaryotic evolutions, oxygenation of the early atmosphere, formation of BIF, and possibly iron-rich extraterrestrial environments.

Derivation of guideline values for gold (III) ion toxicity limits to protect aquatic ecosystems.

PubMed

Nam, Sun-Hwa; Lee, Woo-Mi; Shin, Yu-Jin; Yoon, Sung-Ji; Kim, Shin Woong; Kwak, Jin Il; An, Youn-Joo

2014-01-01

This study focused on estimating the toxicity values of various aquatic organisms exposed to gold (III) ion (Au(3+)), and to propose maximum guideline values for Au(3+) toxicity that protect the aquatic ecosystem. A comparative assessment of methods developed in Australia and New Zealand versus the European Community (EC) was conducted. The test species used in this study included two bacteria (Escherichia coli and Bacillus subtilis), one alga (Pseudokirchneriella subcapitata), one euglena (Euglena gracilis), three cladocerans (Daphnia magna, Moina macrocopa, and Simocephalus mixtus), and two fish (Danio rerio and Oryzias latipes). Au(3+) induced growth inhibition, mortality, immobilization, and/or developmental malformations in all test species, with responses being concentration-dependent. According to the moderate reliability method of Australia and New Zealand, 0.006 and 0.075 mg/L of guideline values for Au(3+) were obtained by dividing 0.33 and 4.46 mg/L of HC5 and HC50 species sensitivity distributions (SSD) with an FACR (Final Acute to Chronic Ratio) of 59.09. In contrast, the EC method uses an assessment factor (AF), with the 0.0006 mg/L guideline value for Au(3+) being divided with the 48-h EC50 value for 0.60 mg/L (the lowest toxicity value obtained from short term results) by an AF of 1000. The Au(3+) guideline value derived using an AF was more stringent than the SSD. We recommend that more toxicity data using various bioassays are required to develop more accurate ecological risk assessments. More chronic/long-term exposure studies on sensitive endpoints using additional fish species and invertebrates not included in the current dataset will be needed to use other derivation methods (e.g., US EPA and Canadian Type A) or the "High Reliability Method" from Australia/New Zealand. Such research would facilitate the establishment of guideline values for various pollutants that reflect the universal effects of various pollutants in aquatic ecosystems. To the best of our knowledge, this is the first study to suggest guideline values for Au(3+) levels permitted to enter freshwater environments. Copyright © 2013 Elsevier Ltd. All rights reserved.

PAR and UV effects on vertical migration and photosynthesis in Euglena gracilis.

PubMed

Richter, Peter; Helbling, Walter; Streb, Christine; Häder, Donat-P

2007-01-01

Recently it was shown that the unicellular flagellate Euglena gracilis changes the sign of gravitaxis from negative to positive upon excessive radiation. This sign change persists in a cell culture for hours even if subsequently transferred to dim light. To test the ecological relevance of this behavior, a vertical column experiment was performed (max. depth 65 cm) to test distribution, photosynthetic efficiency and motility in different horizons of the column (surface, 20, 40 and 65 cm). One column was covered with a UV cut-off filter, which transmits photosynthetically active radiation (PAR) only, the other with a filter which transmits PAR and UV. The columns were irradiated with a solar simulator (PAR 162 W m(-2), UV-A 32.6 W m(-2), UV-B 1.9 W m(-2)). The experiment was conducted for 10 days, normally with a light/dim light cycle of 12 h:12 h, but in some cases the light regime was changed (dim light instead of full radiation). Under irradiation the largest fraction of cells was found at the bottom of the column. The cell density decreased toward the surface. Photosynthetic efficiency, determined with a pulse amplitude modulated fluorometer, was negligible at the surface and increased toward the bottom. While the cell suspension showed a positive gravitaxis at the bottom, the cells in the 40 cm horizon were bimodally oriented (about the same percentage of cells swimming upward and downward, respectively). At 20 cm and at the surface the cells showed negative gravitaxis. Positive gravitaxis was more pronounced in the UV + PAR samples. At the surface and in the 20 and 40 cm horizons photosynthetic efficiency was better in the PAR-only samples than in the PAR + UV samples. At the bottom photosynthetic efficiency was similar in both light treatments. The data suggest that high light reverses gravitaxis of the cells, so that they move downward in the water column. At the bottom the light intensity is lower (attenuation of the water column and self shading of the cells) and the cells recover. After recovery the cells swim upward again until the negative gravitaxis is reversed again.

Graviperception and gravitaxis in flagellates

NASA Astrophysics Data System (ADS)

Häder, D.; Ntefidou, M.

Many photosynthetic and heterotrophic flagellates perceive and respond to the gravitational vector of the Earth. Some previous hypotheses have suggested that the orientation is brought about by a passive physical mechanism such as buoyancy or hydrodynamic alignment. However, recent results have confirmed that e.g. the photosynthetic Euglena utilizes an active physiological sensor and an internal sensory transduction chain. This unicellular organism senses gravity by the sedimentation of its cellular content, which is heavier than the surrounding medium, onto the lower membrane. This force is believed to activate mechano-sensitive ion channels located at the front end under the trailing flagellum. The channels allow a gated influx of calcium which alters the internal electrical potential and may activate calmodulin. Further elements in the transduction chain are cyclic AMP and related enzymes. Recent flight experiments during parabolic aircraft maneuvers and on sounding rockets have confirmed previous terrestrial results and have provided detailed insight into the biochemical sensory transduction chain.

Impact of green algae on the measurement of Microcystis aeruginosa populations in lagoon-treated wastewater with an algae online analyser.

PubMed

Nguyen, Thang; Roddick, Felicity A; Fan, Linhua

2015-01-01

Tests on the algae online analyser (AOA) showed that there was a strong direct linear correlation between cell density and in vivo Chl-a concentration for M. aeruginosa over the range of interest for a biologically treated effluent at a wastewater treatment plant (25,000-65,000 cells mL(-1), equivalent to a biovolume of 2-6 mm3 L(-1)). However, the AOA can provide an overestimate or underestimate of M. aeruginosa populations when green algae are present in the effluent, depending on their species and relative numbers. The results from this study demonstrated that the green algae (e.g., Euglena gracilis, Chlorella sp.) in the field phytoplankton population should be considered during calibration. In summary, the AOA has potential for use as an alert system for the presence of M. aeruginosa, and thus potentially of cyanobacterial blooms, in wastewater stabilization ponds.

Dynamics of Deformable Active Particles under External Flow Field

NASA Astrophysics Data System (ADS)

Tarama, Mitsusuke

2017-10-01

In most practical situations, active particles are affected by their environment, for example, by a chemical concentration gradient, light intensity, gravity, or confinement. In particular, the effect of an external flow field is important for particles swimming in a solvent fluid. For deformable active particles such as self-propelled liquid droplets and active vesicles, as well as microorganisms such as euglenas and neutrophils, a general description has been developed by focusing on shape deformation. In this review, we present our recent studies concerning the dynamics of a single active deformable particle under an external flow field. First, a set of model equations of active deformable particles including the effect of a general external flow is introduced. Then, the dynamics under two specific flow profiles is discussed: a linear shear flow, as the simplest example, and a swirl flow. In the latter case, the scattering dynamics of the active deformable particles by the swirl flow is also considered.

Phytobenthos and phytoplankton community changes upon exposure to a sunflower oil spill in a South African protected freshwater wetland.

PubMed

Oberholster, Paul J; Blaise, Christian; Botha, A-M

2010-11-01

The occurrence of a sunflower oil spill in 2007 in the Con Joubert Bird Sanctuary freshwater wetland, South Africa, inhibited the growth of sensitive phytoplankton species and promoted that of tolerant species. The algal divisions Chlorophyta and Euglenophyta were well represented in the sunflower oil contaminated water, especially the species Euglena sociabilis, Phacus pleuronectes and Chlamydomonas africana. Young and mature resting zygotes of Chlamydomonas africana were recorded in high abundance at all the sunflower oil contaminated sampling sites. The phytobenthos diversity and abundance were significantly suppressed and negatively associated with low Dissolved Oxygen concentrations and the negative redox potential of the bottom sediment. At the intracellular level, phytoplankton chlorophyll a and b concentrations as physiological variables were more sensitive indicators of the adverse effects of sunflower oil than the 72 h Selenastrum capricornutum algal bioassay conducted.

Phytoplankton Functional Groups Variation and Influencing Factors in a Shallow Temperate Lake.

PubMed

Tian, Chang; Hao, Daping; Pei, Haiyan; Doblin, Martina A; Ren, Ying; Wei, Jielin; Feng, Yawei

2018-06-01

  The present study was carried out in Luoma Lake, a shallow lake in temperate eastern China. Based on a two-year study, the dynamics of phytoplankton functional groups and influencing factors were analyzed. A total of 178 taxa were identified and sorted into 20 codons, according to the phytoplankton functional group classification. In order to find the environmental factors driving phytoplankton variations, fifteen groups were analyzed in detail using redundancy analysis. Groups P (Fragilaria crotonensis), X2 (Chlamydomonas globosa, C. microsphaera and Chroomonas acuta), and MP (Navicula rotaeana) were dominant during low temperature periods, whereas groups X2, S1 (Pseudanabaena limnetica), and W1 (Euglena sp.) were dominant during high temperature periods. Water temperature, total phosphorus, and ammonium were the significant driving factors explaining phytoplankton succession. Furthermore, total phosphorus and ammonium could be broadly used in risk management for potential algal blooms in Luoma Lake.

Identification of a volatile phytotoxin from algae

NASA Technical Reports Server (NTRS)

Garavelli, J. S.; Fong, F.; Funkhouser, E. A.

1984-01-01

The objectives were to develop a trap system for isolating fractions of volatile algal phytotoxin and to characterize the major components of the isolated phytotoxin fractions. A bioassay using Phaseolus vulgaris seedlings was developed to aid in investigating the properties of the phytotoxin produced by cultures of Euglena gracilis var. bacillaris and Chlorella vulgaris. Two traps were found, 1.0 M hydrochloric acid and 0 C, which removed the phytotoxin from the algal effluent and which could be treated to release that phytotoxin as judged with the bioassay procedure. It was also determined that pretraps of 1.0 M sodium hydroxide and 1.0 M potassium biocarbonate could be used without lowering the phytotoxin effect. Ammonia was identified in trap solutions by ninhydrin reaction, indophenol reaction and derivatization with dansyl chloride and phenylisothiocyanate. Ammonia at the gaseous concentrations detected was found to have the same effects in the bioassay system as the volatile phytotoxin. It is possible that other basic, nitrogen containing compounds which augment the effects of ammonia were present at lower concentrations in the algal effluent.

Graviperception and gravitaxis in flagellates

NASA Astrophysics Data System (ADS)

Häder, D.-P.; Richter, P.; Ntefidou, M.; Lebert, M.

Unicellular flagellates perceive and react to the gravitational vector of the Earth. Previous hypotheses have suggested that the orientation is brought about by a passive physical mechanism such as buoyancy or hydrodynamic alignment. Recent results of experiments on parabolic rocket flights have revealed that in the photosynthetic Euglena only 10 % of the orientation can be explained by passive orientation while the remainder relies on an active physiological sensor and an internal sensory transduction chain. The cellular contents is heavier than the surrounding medium and consequently presses onto the lower membrane where it activates mechano-sensitive ion channels located at the front end under the trailing flagellum. These channels allow a gated influx of calcium (visualized by confocal microscopy) which depolarizes the internal electrical potential and eventually causes a course correction by the flagellar beating. Further elements in the transduction chain are cyclic AMP and related enzymes. Recent experiments during parabolic aircraft flights and on sounding rockets have confirmed this hypothesis and provided detailed insight into the biochemical sensory transduction chain. Currently the molecular mechanisms of graviperception are being studied.

The physics of the unconventional motility strategy of euglenids

NASA Astrophysics Data System (ADS)

Arroyo, Marino; Noselli, Giovanni; Desimone, Antonio

Euglenids are a family of unicellular protists, which use flagella to move in a fluid. However, they are also capable of performing elegantly concerted large amplitude deformations of the cell shape, in what is known as metaboly. To perform metaboly, euglenids use an elaborate cortical complex capable of actively imposing spatially modulated shear deformations on the cell surface. This mode of cell deformation has been linked to motility, but biophysical studies have demonstrated that it leads to very small swimming velocities as compared to flagellar locomotion. Furthermore, why would these cells possess two elaborate apparatus for the same function remains unclear. In this work, we combine experimental observations of euglena gracilis cells with theoretical models to shed light into the function of metaboly. The theoretical models account for the force generation and shape evolution at the cell envelop, together with the mechanical interaction of the cell with its environment. We characterize the efficiency of the two modes of locomotion of this cells in terms of the physical nature of their environment. ERC AdG 340685 MicroMotility.

Ecotoxicity evaluation of a liquid detergent using the automatic biotest ECOTOX.

PubMed

Azizullah, Azizullah; Richter, Peter; Ullah, Waheed; Ali, Imran; Häder, Donat-Peter

2013-08-01

Synthetic detergents are common pollutants reaching aquatic environments in different ways after usage at homes, institutions and industries. In this study a liquid detergent, used for dish washing, was evaluated for its toxicity during long- and short-term tests using the automatic biotest ECOTOX. Different parameters of Euglena gracilis like motility, swimming velocity, gravitactic orientation, cell compactness and cell growth were used as end points. In short-term experiments, the maximum adverse effects on motility, velocity, cell shape and gravitaxis were observed after 1 h of exposure. With further increase in exposure time to the detergent a slight recovery of these parameters was observed. In long-term experiments, the detergent caused severe disturbances to E. gracilis. Motility, cell growth and cell compactness (shape) with EC50 values of 0.064, 0.18 and 2.05 %, respectively, were found as the most sensitive parameters to detergent stress. There was a slight positive effect on gravitactic orientation at the lowest two concentrations; at higher concentrations of the detergent cells orientation was highly impaired giving EC50 values of 1.75 and 2.52 % for upward swimming and r-value, respectively.

LudusScope: Accessible Interactive Smartphone Microscopy for Life-Science Education.

PubMed

Kim, Honesty; Gerber, Lukas Cyrill; Chiu, Daniel; Lee, Seung Ah; Cira, Nate J; Xia, Sherwin Yuyang; Riedel-Kruse, Ingmar H

2016-01-01

For centuries, observational microscopy has greatly facilitated biology education, but we still cannot easily and playfully interact with the microscopic world we see. We therefore developed the LudusScope, an accessible, interactive do-it-yourself smartphone microscopy platform that promotes exploratory stimulation and observation of microscopic organisms, in a design that combines the educational modalities of build, play, and inquire. The LudusScope's touchscreen and joystick allow the selection and stimulation of phototactic microorganisms such as Euglena gracilis with light. Organismal behavior is tracked and displayed in real time, enabling open and structured game play as well as scientific inquiry via quantitative experimentation. Furthermore, we used the Scratch programming language to incorporate biophysical modeling. This platform is designed as an accessible, low-cost educational kit for easy construction and expansion. User testing with both teachers and students demonstrates the educational potential of the LudusScope, and we anticipate additional synergy with the maker movement. Transforming observational microscopy into an interactive experience will make microbiology more tangible to society, and effectively support the interdisciplinary learning required by the Next Generation Science Standards.

LudusScope: Accessible Interactive Smartphone Microscopy for Life-Science Education

PubMed Central

Kim, Honesty; Gerber, Lukas Cyrill; Chiu, Daniel; Lee, Seung Ah; Cira, Nate J.; Xia, Sherwin Yuyang; Riedel-Kruse, Ingmar H.

2016-01-01

For centuries, observational microscopy has greatly facilitated biology education, but we still cannot easily and playfully interact with the microscopic world we see. We therefore developed the LudusScope, an accessible, interactive do-it-yourself smartphone microscopy platform that promotes exploratory stimulation and observation of microscopic organisms, in a design that combines the educational modalities of build, play, and inquire. The LudusScope’s touchscreen and joystick allow the selection and stimulation of phototactic microorganisms such as Euglena gracilis with light. Organismal behavior is tracked and displayed in real time, enabling open and structured game play as well as scientific inquiry via quantitative experimentation. Furthermore, we used the Scratch programming language to incorporate biophysical modeling. This platform is designed as an accessible, low-cost educational kit for easy construction and expansion. User testing with both teachers and students demonstrates the educational potential of the LudusScope, and we anticipate additional synergy with the maker movement. Transforming observational microscopy into an interactive experience will make microbiology more tangible to society, and effectively support the interdisciplinary learning required by the Next Generation Science Standards. PMID:27706189

The application of Fresnel zone plate based projection in optofluidic microscopy.

PubMed

Wu, Jigang; Cui, Xiquan; Lee, Lap Man; Yang, Changhuei

2008-09-29

Optofluidic microscopy (OFM) is a novel technique for low-cost, high-resolution on-chip microscopy imaging. In this paper we report the use of the Fresnel zone plate (FZP) based projection in OFM as a cost-effective and compact means for projecting the transmission through an OFM's aperture array onto a sensor grid. We demonstrate this approach by employing a FZP (diameter = 255 microm, focal length = 800 microm) that has been patterned onto a glass slide to project the transmission from an array of apertures (diameter = 1 microm, separation = 10 microm) onto a CMOS sensor. We are able to resolve the contributions from 44 apertures on the sensor under the illumination from a HeNe laser (wavelength = 633 nm). The imaging quality of the FZP determines the effective field-of-view (related to the number of resolvable transmissions from apertures) but not the image resolution of such an OFM system--a key distinction from conventional microscope systems. We demonstrate the capability of the integrated system by flowing the protist Euglena gracilis across the aperture array microfluidically and performing OFM imaging of the samples.

Effect of long-term microgravity on the mineralisation of inner ear otoliths of fish - a spaceflight study

NASA Astrophysics Data System (ADS)

Anken, Ralf

The "heavy bodies" (i.e., statoliths or otoliths, mainly made up of calcium carbonate and protein) in the inner ears of vertebrates transform the physical parameter "gravity" to biological signals needed for postural control. It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish (via a down-regulation of carbonic anhydrase reactivity) as an adaptation towards altered environmental gravity. We were thus prompted to elucidate whether long-term microgravity would possibly yield opposite effects. Therefore, larval siblings of cichlid fish (Oreochromis mossambicus) were housed in a bioregenerative life support system (OMEGAHAB) using green algae (Euglena gracilis) for oxygen supply. The experiment was successfully flown on the FOTON M-3 mission. Prior to launch, otoliths were stained with a fluorescent calcium tracer (Alizarin Complexone). This treatment both allowed an assessment of otolith growth (size) after recovery as well as an analysis of relocations of calcium deposits. Calcium and strontium contents were determined using inductively coupled plasma mass spectrometry. The results will be communicated at the meeting. Acknowledgement: This work was financially supported by the German Aerospace Center (DLR) (FKZ: 50 WB 0527).

Phytoplankton species diversity indices in Anjanapura reservoir, Western Ghat region, India.

PubMed

Basavaraja, D; Narayana, J; Puttaiah, E T; Prakash, K

2013-07-01

A qualitative study on species composition and diversity of phytoplankton flora was carried out in the Anjanapura reservoir. Water samples were collected monthly at four sites from November 2005 to October 2007 for plankton analysis. A total of 152 phytoplankton species belonging to 59 genera were recorded. The species belonging to genera Crucigenia, Pediastrum, Scenedesmus, Tetraedron, Cyclotella, Gyrosigma, Melosira, Navicula, Fragillaria, pinnularia, Synedra, Closterium, Cosmarium, Euastrum, Staurastrum, Gleocapsa, Merismopedia, Microcystis, Oscillatoria, Euglena and Phacus were found dominant. Phytoplankton density was recorded in the range of 222-1467 Org l(-1) for chlorococcales, 267-1467 Org I (-1) for diatoms, 44-889 Org l(-1) for desmids, 44-889 Org l(-1) for blue-greens and 22-1667 Org l(-1) for euglenoids. Seasonal density Shannon-Weiner diversity index (1.60-1.88) indicate that reservoir exhibited rich phytoplankton and Nygaard tropic status indices for myxophycean (1.0-1.50), chlorophycean (1.62-2.16), diatoms (0.10-0.14), euglenophycean (0.18-0.19) and compound quotient(3.25-4.66) showed that reservoir was moderately polluted. From the observed data, it could be concluded that the presence of more diverse group of phytoplankton indicate the productivity of the reservoir.

Evolutionary History of the Enzymes Involved in the Calvin-Benson Cycle in Euglenids.

PubMed

Markunas, Chelsea M; Triemer, Richard E

2016-05-01

Euglenids are an ancient lineage that may have existed as early as 2 billion years ago. A mere 65 years ago, Melvin Calvin and Andrew A. Benson performed experiments on Euglena gracilis and elucidated the series of reactions by which carbon was fixed and reduced during photosynthesis. However, the evolutionary history of this pathway (Calvin-Benson cycle) in euglenids was more complex than Calvin and Benson could have imagined. The chloroplast present today in euglenophytes arose from a secondary endosymbiosis between a phagotrophic euglenid and a prasinophyte green alga. A long period of evolutionary time existed before this secondary endosymbiotic event took place, which allowed for other endosymbiotic events or gene transfers to occur prior to the establishment of the green chloroplast. This research revealed the evolutionary history of the major enzymes of the Calvin-Benson cycle throughout the euglenid lineage and showed that the majority of genes for Calvin-Benson cycle enzymes shared an ancestry with red algae and/or chromophytes suggesting they may have been transferred to the nucleus prior to the acquisition of the green chloroplast. © 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.

Effects of acid mine drainage on the stream ecosystem of the east fork of the Obey River, Tennessee

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nichols, L.E.; Bulow, F.L.

1973-01-01

The stream ecosystem of the east fork of the Obey River, Tennessee was studied from January through December 1970. Emphasis centered on water quality, macroinvertebrates, fish and aquatic flora affected by acid mine drainage. Two control stations were established within the study area, one located below the zone of pollution. A reservoir station was established to detect any neutralization occurring within Dale Hollow Reservoir below the confluence of the east fork and west fork. An area approximately 40 miles in length was found to be severely degraded by acid mine drainage. Limited macroinvertebrate populations existed within this region. Chironomus andmore » Sialis were the predominate benthic indicator organisms present in the polluted zone. Euglena mutabilis was the most abundant representative organism of the aquatic flora. This organism was found to be characteristic of acid mine pollution. Fish were recovered above and below, but not within, the zone of pollution. Fish recovered were characteristic of the type of habitat in which they were collected, being either typical stream or reservoir fish. A fish kill occurred in the east fork embayment of Dale Hollow on August 15, 1970 from acid mine drainage discharged upstream.« less

Differences in planktonic microbial communities associated with three types of macrophyte stands in a shallow lake.

PubMed

Mentes, Anikó; Szabó, Attila; Somogyi, Boglárka; Vajna, Balázs; Tugyi, Nóra; Csitári, Bianka; Vörös, Lajos; Felföldi, Tamás

2018-02-01

Little is known about how various substances from living and decomposing aquatic macrophytes affect the horizontal patterns of planktonic bacterial communities. Study sites were located within Lake Kolon, which is a freshwater marsh and can be characterised by open-water sites and small ponds with different macrovegetation (Phragmites australis, Nymphea alba and Utricularia vulgaris). Our aim was to reveal the impact of these macrophytes on the composition of the planktonic microbial communities using comparative analysis of environmental parameters, microscopy and pyrosequencing data. Bacterial 16S rRNA gene sequences were dominated by members of phyla Proteobacteria (36%-72%), Bacteroidetes (12%-33%) and Actinobacteria (5%-26%), but in the anoxic sample the ratio of Chlorobi (54%) was also remarkable. In the phytoplankton community, Cryptomonas sp., Dinobryon divergens, Euglena acus and chrysoflagellates had the highest proportion. Despite the similarities in most of the measured environmental parameters, the inner ponds had different bacterial and algal communities, suggesting that the presence and quality of macrophytes directly and indirectly controlled the composition of microbial plankton. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Interactive Biophysics with Microswimmers: Education, Cloud Experimentation, Programmed Swarms, and Biotic Games

NASA Astrophysics Data System (ADS)

Riedel-Kruse, Ingmar

Modern biotechnology gets increasingly powerful to manipulate and measure microscopic biophysical processes. Nevertheless, no platform exists to truly interact with these processes, certainly not with the convenience that we are accustomed to from our electronic smart devices. In my talk I will provide the rational for such Interactive Biotechnology and conceptualize its core component, the BPU (biotic processing unit), which is then connected to an according user interface. The biophysical phenomena currently featured on these platforms utilize the phototactic response of motile microorganisms, e.g., Euglena gracilis, resulting in spatio-temporal dynamics from the single cell to the self-organized multi-cellular scale. I will demonstrate multiple platforms, such as scalable biology cloud experimentation labs, tangible museum exhibits, biotic video games, low-cost interactive DIY kits using smartphones, and programming languages for swarm robotics. I will discuss applications for education as well as for professional and citizen science. Hence, we turn traditionally observational microscopy into an interactive experience. I was told that presenting in the educational section does not count against the ''one author - one talk policy'' - so I submit two abstracts. In case of conflict - please contact me: ingmar@stanford.edu.

Profiling planktonic biomass using element-specific, multicomponent nuclear magnetic resonance spectroscopy.

PubMed

Komatsu, Takanori; Kobayashi, Toshiya; Hatanaka, Minoru; Kikuchi, Jun

2015-06-02

Planktonic metabolism plays crucial roles in Earth's elemental cycles. Chemical speciation as well as elemental stoichiometry is important for advancing our understanding of planktonic roles in biogeochemical cycles. In this study, a multicomponent solid-state nuclear magnetic resonance (NMR) approach is proposed for chemical speciation of cellular components, using several advanced NMR techniques. Measurements by ssNMR were performed on (13)C and (15)N-labeled Euglena gracilis, a flagellated protist. 3D dipolar-assisted rotational resonance, double-cross-polarization (1)H-(13)C correlation spectroscopy, and (1)H-(13)C solid-state heteronuclear single quantum correlation spectroscopy successively allowed characterization of cellular components. These techniques were then applied to E. gracilis cultured in high and low ammonium media to demonstrate the power of this method for profiling and comparing cellular components. Cellular NMR spectra indicated that ammonium induced both paramylon degradation and amination. Arginine was stored as a nitrogen reserve and ammonium replaced by arginine catabolism via the arginine dihydrolase pathway. (15)N and (31)P cellular ssNMR indicated arginine and polyphosphate accumulation in E. gracilis, respectively. This chemical speciation technique will contribute to environmental research by providing detailed information on environmental chemical properties.

Development of phytotoxicity tests using wetland species

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nelson, M.K.; Fairchild, J.F.

1994-12-31

Laboratory phytotoxicity tests used to assess contaminant effects may not effectively protect wetland communities. The authors are developing routine culture and testing methods for selected fresh water plants, that can be used in risk assessments and monitoring of existing wetland systems. Utility of these tests includes evaluating the effects of point or non-point source contamination that may cause water or sediment quality degradation. Selected species include algae (blue-green, green), phytoflagellates (Chlamydomonas, Euglena), and floating or submerged vascular plants (milfoil, coontail, wild celery, elodea, duckweed). Algae toxicity tests range from 2-d, 4-d, and 7 day tests, and macrophyte tests from 10-dmore » to 14 days. Metribuzin and boron are the selected contaminants for developing the test methods. Metribuzin, a triazinone herbicide, is a photosystem 11 inhibitor, and is commonly used for control of grass and broad-leaf plants. As a plant micronutrient, boron is required in very small amounts, but excessive levels can result in phytotoxicity or accumulation. The investigations focus on the influence of important factors including the influence of light quality and quantity, and nutrient media. Reference toxicant exposures with potassium chloride are used to establish baseline data for sensitivity and vitality of the plants. These culture and test methods will be incorporated into recommendations for standard phytotoxicity test designs.« less

Immunochemical Studies on α-Amylase III. Immunochemical Relationships Among Amylases from Various Microorganisms1

PubMed Central

Sirisinha, Stitaya; Allen, Peter Z.

1965-01-01

Sirishinha, Stitaya (University of Rochester School of Medicine and Dentistry, Rochester, N.Y.), and Peter Z. Allen. Immunochemical studies on α-amylase. III. Immunochemical relationships among amylases from various microorganisms. J. Bacteriol. 90:1120–1128. 1965.—Immunochemical relationships among amylases obtained from a selected group of microorganisms were examined, and a cross-reaction was detected between the α-amylases of Bacillus stearothermophilus and B. subtilis. Immunodiffusion and quantitative precipitin studies, as well as cross-neutralization tests, indicate that B. stearothermophilus α-amylase reacts with a portion of antibody present in antisera to crystalline B. subtilis α-amylase. Amylases from these two species thus have some aspects of structure in common. Limited data obtained by immunodiffusion suggest that groupings which confer cross-reactivity to the B. stearothermophilus enzyme are lost after exposure to mercaptoethanol in the presence of ethylenediamine-tetraacetate, followed by treatment with iodoacetamide. With the antisera employed and within the concentration range examined, no immunochemical cross-reaction was observed among amylases from Aspergillus oryzae, B. subtilis, B. polymyxa, B. macerans, Pseudomonas saccharophila, and Euglena sanguinis. Immunoelectrophoresis of partially purified B. stearothermophilus α-amylase by use of antiserum to the crude enzyme, together with localization of amylase activity in immunoelectrophoretic plates, suggests that B. stearothermophilus α-amylase is antigenic in the rabbit. Images PMID:5847799

Diverse strategies of O2 usage for preventing photo-oxidative damage under CO2 limitation during algal photosynthesis.

PubMed

Shimakawa, Ginga; Matsuda, Yusuke; Nakajima, Kensuke; Tamoi, Masahiro; Shigeoka, Shigeru; Miyake, Chikahiro

2017-01-20

Photosynthesis produces chemical energy from photon energy in the photosynthetic electron transport and assimilates CO 2 using the chemical energy. Thus, CO 2 limitation causes an accumulation of excess energy, resulting in reactive oxygen species (ROS) which can cause oxidative damage to cells. O 2 can be used as an alternative energy sink when oxygenic phototrophs are exposed to high light. Here, we examined the responses to CO 2 limitation and O 2 dependency of two secondary algae, Euglena gracilis and Phaeodactylum tricornutum. In E. gracilis, approximately half of the relative electron transport rate (ETR) of CO 2 -saturated photosynthesis was maintained and was uncoupled from photosynthesis under CO 2 limitation. The ETR showed biphasic dependencies on O 2 at high and low O 2 concentrations. Conversely, in P. tricornutum, most relative ETR decreased in parallel with the photosynthetic O 2 evolution rate in response to CO 2 limitation. Instead, non-photochemical quenching was strongly activated under CO 2 limitation in P. tricornutum. The results indicate that these secondary algae adopt different strategies to acclimatize to CO 2 limitation, and that both strategies differ from those utilized by cyanobacteria and green algae. We summarize the diversity of strategies for prevention of photo-oxidative damage under CO 2 limitation in cyanobacterial and algal photosynthesis.

Distribution of biota in a stream polluted by acid mine-drainage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Warner, R.W.

1971-01-01

Acidic water draining from coal mines has severly restricted the diversity of biota inhabiting Roaring Creek, eastern West Virginia. Polluted reaches of the stream (median pH values ranging from 2.8 to 3.8) were inhabited by 3 to 12 genera of bottom-dwelling invertebrates and 10 to 19 species of periphytic algae. Invertebrates tolerant of the pollution included Sialis sp., chironomus plumosus and other Chironomidae, dytiscid beetles, and Ptilostomis sp. Predominant among the tolerant periphyton were Ulothrix tenerrima, Pinnularia termitina, Eunotia exigua, and Euglena mutabilis. Six other species of algae were tolerant of the acid mine-pollution, but were never numerous. Sections ofmore » Roaring Creek not severely polluted by acid drainage (pH medians of 4.5 or higher) supported diverse communities of 25 or more kinds of benthic animals and 27 or more species of periphytic algae. These stream reaches were inhibited by blackflies, crayfish, mayflies, stoneflies, and many species of caddisflies; these forms did not inhabit the more acidic stream reaches. Because of the complex and varying chemical composition of the acid mine-drainage, and also because of possible physical influences, measurements of pH values in the stream seemed to provide the most reliable, as well as unique, index of the effects of acid mine-drainage on aquatic life.« less

Assessing Transformations of Algal Organic Matter in the Long-Term: Impacts of Humification-Like Processes

PubMed Central

Leloup, Maud; Pallier, Virginie; Nicolau, Rudy; Feuillade-Cathalifaud, Geneviève

2015-01-01

Algae and cyanobacteria are important contributors to the natural organic matter (NOM) of eutrophic water resources. The objective of this work is to increase knowledge on the modifications of algal organic matter (AOM) properties in the long term to anticipate blooms footprint in such aquatic environments. The production of AOM from an alga (Euglena gracilis) and a cyanobacteria (Microcystis aeruginosa) was followed up and characterized during the stationary phase and after one year and four months of cultivation, in batch experiments. Specific UV absorbance (SUVA) index, organic matter fractionation according to hydrophobicity and apparent molecular weight were combined to assess the evolution of AOM. A comparison between humic substances (HS) mainly derived from allochthonous origins and AOM characteristics was performed to hypothesize impacts of AOM transformation processes on the water quality of eutrophic water resources. Each AOM fraction underwent a specific evolution pattern, depending on its composition. Impacts of humification-like processes were predominant over release of biopolymers due to cells decay and led to an increase in the hydrophobic compounds part and molecular weights over time. However, the hydrophilic fraction remained the major fraction whatever the growth stage. Organic compounds generated by maturation of these precursors corresponded to large and aliphatic structures. PMID:26251898

Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution.

PubMed

Maeno, Takanori; Uzawa, Takanori; Kono, Izumi; Okano, Kazunori; Iino, Takanori; Fukita, Keisuke; Oshikawa, Yuki; Ogawa, Taro; Iwata, Osamu; Ito, Takuro; Suzuki, Kengo; Goda, Keisuke; Hosokawa, Yoichiroh

2018-05-29

Microalgae-based metabolic engineering has been proven effective for producing valuable substances such as food supplements, pharmaceutical drugs, biodegradable plastics, and biofuels in the past decade. The ability to accurately visualize and quantify intracellular metabolites in live microalgae is essential for efficient metabolic engineering, but remains a major challenge due to the lack of characterization methods. Here we demonstrate it by synthesizing fluorogenic peptide aptamers with specific binding affinity to a target metabolite and delivering them into live microalgae by femtosecond laser photoporation at single-cell resolution. As a proof-of-principle demonstration of our method, we use it to characterize Euglena gracilis, a photosynthetic unicellular motile microalgal species, which is capable of producing paramylon (a carbohydrate granule similar to starch). Specifically, we synthesize a peptide aptamer containing a paramylon-binding fluorescent probe, 7-nitrobenzofurazan, and introduce it into E. gracilis cells one-by-one by suppressing their mobility with mannitol and transiently perforating them with femtosecond laser pulses at 800 nm for photoporation. To demonstrate the method's practical utility in metabolic engineering, we perform spatially and temporally resolved fluorescence microscopy of single live photoporated E. gracilis cells under different culture conditions. Our method holds great promise for highly efficient microalgae-based metabolic engineering.

[Studies on the role of silicic acid in the development of higher plants].

PubMed

Werner, D

1967-03-01

Germanium acid, a specific inhibitor of the silicic acid metabolism in diatoms, inhibits the growth of Sinapis alba, Lemna minor, Wolffia arrhiza, Nicotiana tabacum, Tradescantia spec, Zinnia elegans, and Secale cereale when applied in the same concentrations as those used in the case of diatoms (15-75 μg GeO2/ml medium). The growth of Aspergillus niger, Phycomyces blakesleanus, Escherichia coli K 12, Euglena gracilis and Pandorina morum is not influenced by these and higher concentrations of Germanium acid. By application of high concentrations of silicic acid, the growth inhibition produced by germanium acid in Lemna minor is partially reduced. Plants of Lemna minor which have been inhibited by germanium acid are essentially smaller than plants grown in a normal medium; their chlorophyll content is significantly decreased. The growth of the roots in Lemna is particularly inhibited. Isolated growing roots of Lycopersicon pimpinellifolium MILL. are inhibited by small concentrations of Ge(OH)4 (ca. 1,5×10(-4) M/l). In contrast to the growth of older plants, the germination of Secale cereale and Sinapis alba is not influenced by Ge(OH)4. The effects of germanium acid are discussed in relation to the physiological role of silicic acid. The results suggest that the element silicon, in the form of silicic acid, is generally essential for the normal development of higher plants.

Species selection for the design of gold nanobioreactor by photosynthetic organisms

NASA Astrophysics Data System (ADS)

Dahoumane, Si Amar; Djediat, Chakib; Yéprémian, Claude; Couté, Alain; Fiévet, Fernand; Coradin, Thibaud; Brayner, Roberta

2012-06-01

The design of cell-based bioreactors for inorganic particles formation requires both a better understanding of the underlying processes and the identification of most suitable organisms. With this purpose, the process of Au3+ incorporation, intracellular reduction, and Au0 nanoparticle release in the culture medium was compared for four photosynthetic microorganisms, Klebsormidium flaccidum and Cosmarium impressulum green algae, Euglena gracilis euglenoid and Anabaena flos- aquae cyanobacteria. At low gold content, the two green algae show maintained photosynthetic activity and recovered particles (ca. 10 nm in size) are similar to internal colloids, indicating a full biological control over the whole process. In similar conditions, the euglenoid exhibits a rapid loss of biological activity, due to the absence of protective extracellular polysaccharide, but could grow again after an adaptation period. This results in a larger particle size dispersity but larger reduction yield. The cyanobacteria undergo rapid cell death, due to their prokaryotic nature, leading to high gold incorporation rate but poor control over released particle size. Similar observations can be made after addition of a larger gold salt concentration when all organisms rapidly die, suggesting that part of the process is not under biological control anymore but also involves extracellular chemical reactions. Overall, fruitful information on the whole biocrystallogenesis process is gained and most suitable species for further bioreactor design can be identified, i.e., green algae with external coating.

A More Desirable Balanced Polyunsaturated Fatty Acid Composition Achieved by Heterologous Expression of Δ15/Δ4 Desaturases in Mammalian Cells

PubMed Central

Zhu, Guiming; Ou, Qin; Zhang, Tao; Jiang, Xudong; Sun, Guozhi; Zhang, Ning; Wang, Kunfu; Fang, Heng; Wang, Mingfu; Sun, Jie; Ge, Tangdong

2013-01-01

Arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids are the most biologically active polyunsaturated fatty acids, but their biosyntheses in mammals are very limited. The biosynthesis of DHA is the most difficult, because this undergoes the Sprecher pathway–a further elongation step from docosapentaenoic acid (DPA), a Δ6-desaturase acting on a C24 fatty acid substrate followed by a peroxisomal chain shortening step. This paper reports the successful heterologous expression of two non-mammalian genes (with modification of codon usage), coding for Euglena gracilis Δ4-desaturase and Siganus canaliculatus Δ4-desaturase respectively, in mammalian cells (HEK293 cell line). Both of the Δ4-desaturases can efficiently function, directly converting DPA into DHA. Moreover, the cooperation of the E. gracilis Δ4-desaturase with C. elegans Δ15-desaturase (able to convert a number of n-6 PUFAs to their corresponding n-3 PUFAs) in transgenic HEK293 cells made a more desirable fatty acid composition – a drastically reduced n-6/n-3 PUFAs ratio and a high level of DHA as well as EPA and ARA. Our findings provide a basis for potential applications of the gene constructs for expression of Δ15/Δ4-desaturases in transgenic livestock to produce such a fatty acid profile in the related products, which certainly will bring benefit to human health. PMID:24391980

Hunting for agile prey: trophic specialisation in leptophryid amoebae (Vampyrellida, Rhizaria) revealed by two novel predators of planktonic algae.

PubMed

Hess, Sebastian

2017-09-01

Vampyrellid amoebae (Vampyrellida, Rhizaria) are widespread in freshwater, marine and terrestrial ecosystems and consume a wide range of eukaryotes, e.g. algae, fungi and micrometazoa. Environmental sequences indicate that only a small fraction of their genetic diversity is phenotypically characterised, emphasising the need to further explore unknown vampyrellids and their interactions with prey organisms. This study tests the prey range specificity of three vampyrellid amoebae with 49 strains of three common groups of freshwater algae (Zygnematophyceae, Euglenophyceae and Volvocales), and documents specific interactions by time-lapse microscopy. Two of the amoebae, here introduced as the novel genera Arachnomyxa and Planctomyxa based on morphology and SSU rRNA gene comparisons, display a complementary prey range and consume motile algae, namely Volvocales and Euglenophyceae, respectively. This reveals the existence of specialised 'plankton feeders' in the vampyrellid family Leptophryidae, contrasting with the strikingly broad prey range of Leptophrys vorax. The distinct autecological characteristics found in this group of morphologically rather indistinct amoebae contribute to our knowledge about the vastly understudied vampyrellid amoebae. Furthermore, time-lapse observations suggest that euglenoid movements exerted by the sluggish species of the 'Euglena deses group' as a reaction to vampyrellid contact may serve as an effective defence against microbial predators. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Modul.LES: a multi-compartment, multi-organism aquatic life support system as experimental platform for research in ∆g

NASA Astrophysics Data System (ADS)

Hilbig, Reinhard; Anken, Ralf; Grimm, Dennis

In view of space exploration and long-term satellite missions, a new generation of multi-modular, multi-organism bioregenerative life support system with different experimental units (Modul.LES) is planned, and subunits are under construction. Modul.LES will be managed via telemetry and remote control and therefore is a fully automated experimental platform for different kinds of investigations. After several forerunner projects like AquaCells (2005), C.E.B.A.S. (1998, 2003) or Aquahab (OHB-System AG the Oreochromis Mossambicus Eu-glena Gracilis Aquatic Habitat (OmegaHab) was successfully flown in 2007 in course of the FOTON-M3 Mission. It was a 3 chamber controlled life support system (CLSS), compris-ing a bioreactor with the green algae Euglena gracilis, a fish chamber with larval cichlid fish Oreochromis mossambicus and a filter chamber with biodegrading bacteria. The sensory super-vision of housekeeping management was registered and controlled by telemetry. Additionally, all scientific data and videos of the organisms aboard were stored and sequentially transmitted to relay stations. Based on the effective performance of OmegaHab, this system was chosen for a reflight on Bion-M1 in 2012. As Bion-M1 is a long term mission (appr. 4 weeks), this CLSS (OmegaHab-XP) has to be redesigned and refurbished with enhanced performance. The number of chambers has been increased from 3 to 4: an algae bioreactor, a fish tank for adult and larval fish (hatchery inserted), a nutrition chamber with higher plants and crustaceans and a filter chamber. The OmegaHab-XP is a full automated system with an extended satellite downlink for video monitoring and housekeeping data acquisition, but no uplink for remote control. OmegaHab-XP provides numerous physical and chemical parameters which will be monitored regarding the state of the biological processes and thus enables the automated con-trol aboard. Besides the two basic parameters oxygen content and temperature, products of the nitrogene-cycle (concentration of ammonium, nitrite and nitrate) as well as conductivity will be measured. For this long term mission an external food supply as has been used with OmegaHab is not sufficient and, therefore, in OmegaHab-XP a nutrition compartment has been added. OmegaHab-XP is a multi-trophic system, designed as a basic concept and test-bed for future multi-modular platform Modul.LES. OmegaHab-XP comprises four different trophic lev-els. The algae experimental container is used as CO2 / O2 exchanger and serves as oxygen source for all heterotrophic organisms. The fish compartment is divided into two areas -namely a hatchery (larval cichlid fish Oreochromis mossambicus) and a fish tank (subadult cichlids). Once the yolk sack is resorbed (stage 19) the juvenile fish are capable to leave the hatchery via escapements into the fish compartment. In order to enable the development of fish from larval yolk sack stages to subadult fish a nutrition compartment is enclosed: In this nutrition compartment the crustacean Hyalella azteca will reproduce and build up a stable population by feeding on the Rigid Hornwort (Ceratophyllum demersum). Younger crustaceans can cross the barrier to the fish tank and can serve as nutrition for fully developed subadult fish. Waste products of all organisms will be assimilated by the water snail Biomphalaria glabrata. The scientific concept of Modul.LES is to establish a multidisciplinary framework of scientists and areas of scientific research (biophysics, molecular-organismic biology, biochemistry etc.) to analyze impacts of g on plants and animals.

Phytoplankton abundance and structure as indicator of water quality in the drainage system of the Burullus Lagoon, southern Mediterranean coast, Egypt.

PubMed

El-Kassas, Hala Yassin; Gharib, Samiha Mahmoud

2016-09-01

This study represents the first detailed account of phytoplankton community structure and seasonal succession in eight drain sites and the Brimbal Canal influx into the Burullus Lagoon. The phytoplankton characteristics were studied based on the data collected seasonally over 4 years, from summer 2012 to spring 2016. Various indices such as Palmer's and Shannon's biotic indices were used for the assessment of the water quality of the different drains. There were a total of 194 species belonging to 65 genera and 6 groups: Bacillariophyceae (76 species), Chlorophyceae (59 species), Cyanophyceae (30 species), Euglenophyceae (25 species), Dinophyceae (3 species), and Xanthophyceae (1 species). The phytoplankton community was dominated with diatoms, green algae, and euglenoids such as Cyclotella, Scenedesmus, Navicula, Nitzschia, Ankistrodesmus, Chlorella, and Euglena. Maximum and minimum phytoplankton abundance was recorded at the Brimbal Canal and Hooks Drain. Maximum and minimum species diversities (H') were found at the Hooks Drain (2.564) and Burullus Drain (2.055). Species evenness fluctuated between 0.595 (Burullus Drain) and 0.750 (West Burullus Drain). The total score of algal genus pollution index and the algal species pollution index at the different drains showed that Drain 7 and the West Burullus Drain had moderate pollution, and the total score of the other drains were greater than 20 indicating the confirmed high organic pollution. Thus, the present investigation can be considered an attempt to use the phytoplankton community as a bioindicator of organic pollution.

Advancing Small Satellite Electronics Heritage for Microfluidic Biological Experiments

NASA Technical Reports Server (NTRS)

White, Bruce; Mazmanian, Edward; Tapio, Eric

2016-01-01

DLR's Eu:CROPIS (Euglena and Combined Regenerative Organic-Food Production in Space) mission, launching in 2017, will carry multiple biological payloads into a sun-synchronous orbit, including NASA Ames' PowerCell experiment. PowerCell will attempt to characterize the viability of synthetic biology at micro-g, Lunar, and Martian gravity levels. PowerCell experiment requirements demand an electronic system similar to previous microfluidic biology payloads, but with an expanded feature set. As such, the system was based on PharmaSat (Diaz-Aguado et al. 2009), a previous successful biology payload from NASA Ames, and improved upon. Newer, more miniaturized electronics allow for greater capability with a lower part count and smaller size. Two identical PowerCell enclosures will fly. Each enclosure contains two separate and identical experiments with a 48-segment optical density measurement system, grow light system, microfluidic system for nutrient delivery and waste flushing, plus thermal control and environmental sensing/housekeeping including temperature, pressure, humidity, and acceleration. Electronics consist of a single Master PCB that interfaces to the spacecraft bus and regulates power and communication, plus LED, Detector, and Valve Manifold PCBs for each experiment. To facilitate ease of reuse on future missions, experiment electronics were designed to be compatible with a standard 3U small sat form factor and power bus, or to interface with a Master power/comm PCB for use in a larger satellite as in the case of PowerCell's flight on Eu:CROPIS.

Application of a Depositional Facies Model to an Acid Mine Drainage Site▿ †

PubMed Central

Brown, Juliana F.; Jones, Daniel S.; Mills, Daniel B.; Macalady, Jennifer L.; Burgos, William D.

2011-01-01

Lower Red Eyes is an acid mine drainage site in Pennsylvania where low-pH Fe(II) oxidation has created a large, terraced iron mound downstream of an anoxic, acidic, metal-rich spring. Aqueous chemistry, mineral precipitates, microbial communities, and laboratory-based Fe(II) oxidation rates for this site were analyzed in the context of a depositional facies model. Depositional facies were defined as pools, terraces, or microterracettes based on cm-scale sediment morphology, irrespective of the distance downstream from the spring. The sediments were composed entirely of Fe precipitates and cemented organic matter. The Fe precipitates were identified as schwertmannite at all locations, regardless of facies. Microbial composition was studied with fluorescence in situ hybridization (FISH) and transitioned from a microaerophilic, Euglena-dominated community at the spring, to a Betaproteobacteria (primarily Ferrovum spp.)-dominated community at the upstream end of the iron mound, to a Gammaproteobacteria (primarily Acidithiobacillus)-dominated community at the downstream end of the iron mound. Microbial community structure was more strongly correlated with pH and geochemical conditions than depositional facies. Intact pieces of terrace and pool sediments from upstream and downstream locations were used in flowthrough laboratory reactors to measure the rate and extent of low-pH Fe(II) oxidation. No change in Fe(II) concentration was observed with 60Co-irradiated sediments or with no-sediment controls, indicating that abiotic Fe(II) oxidation was negligible. Upstream sediments attained lower effluent Fe(II) concentrations compared to downstream sediments, regardless of depositional facies. PMID:21097582

Preliminary study on applicability of microsatellite DNA primers from parasite protozoa Trypanosoma cruzi in free-living protozoa

NASA Astrophysics Data System (ADS)

Zhang, Wenjing; Yu, Yuhe; Shen, Yunfen; Miao, Wei; Feng, Weisong

2004-04-01

In this paper, we took the lead in studying on specificity of the microsatellite DNA loci and applicability of microsatellite DNA primers in protozoa. In order to study characters of microsatellites in free-living protozoa, eight microsatellite loci primers developed from Trypanosoma cruzi (MCLE01, SCLE10, MCLE08, SCLE11, MCLF10, MCLG10, MCL03, MCL05) were employed to amplify microsatellite in four free-living protozoa, including Bodo designis, Euglena gracilis FACHB848, Paramecium bruzise and Tetrahymena thermophila BF1. In the amplification systems of P. bruzise, four loci (SCLE10, SCLE11, MCLF10, MCL03) were amplified successfully, and four amplification fragments were in proper size. In genome of E. gracilis FACHB848, five of eight primers brought five clear amplification bands. In B. designis, three (No.4, 5 and 7) of eight loci produced clear and sharp products without stutter bands, whereas no bands appeared in T. thermophila BF1. Further, eight 300 500 bp amplification fragments were cloned and sequenced. Nevertheless, all sequenced products did not contain corresponding microsatellite sequence, although Bodo is in the same order and has the nearest phylogenetic relation with Trypanosoma among these four species. Thus, the microsatellite DNA primers can not be applied among order or more far taxa, and the specificity of microsatellite DNA is very high in protozoa. The results of this study will contribute to our understanding of microsatellite DNA in protozoa.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) coupled to XAD fractionation: Method to algal organic matter characterization.

PubMed

Nicolau, Rudy; Leloup, Maud; Lachassagne, Delphine; Pinault, Emilie; Feuillade-Cathalifaud, Geneviève

2015-05-01

This work is focused on the development of an analytical procedure for the improvement of the Organic Matter structure characterization, particularly the algal matter. Two fractions of algal organic matter from laboratory cultures of algae (Euglena gracilis) and cyanobacteria (Microcystis aeruginosa) were extracted with XAD resins. The fractions were studied using laser desorption ionization (LDI) and Matrix-Assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). A comparison with the natural organic matter characteristics from commercial humic acids and fulvic acids extracted from Suwannee River was performed. Results show that algal and natural organic matters have unique quasi-polymeric structures. Significant repeating patterns were identified. Different fractions extracted from organic matter with common origin had common structures. Thus, 44, 114 and 169Da peaks separation for fractions from E. gracilis organic matter and 28, 58 and 100Da for M. aeruginosa ones were clearly observed. Using the developed protocol, a structural scheme and organic matter composition were obtained. The range 600-2000Da contained more architectural composition differences than the range 100-600Da, suggesting that organic matter is composed of an assembly of common small molecules. Associated to specific monomers, particular patterns were common to all samples but assembly and resulting structure were unique for each organic matter. Thus, XAD fractionation coupled to mass spectroscopy allowed determining a specific fingerprint for each organic matter. Copyright © 2015 Elsevier B.V. All rights reserved.

Effects of chronic γ-irradiation on the aquatic microbial microcosm: equi-dosimetric comparison with effects of heavy metals.

PubMed

Fuma, Shoichi; Kawaguchi, Isao; Kubota, Yoshihisa; Yoshida, Satoshi; Kawabata, Zen'ichiro; Polikarpov, Gennady G

2012-02-01

Effects of chronic γ-irradiation were investigated in the aquatic microcosm consisting of flagellate algae Euglena gracilis as producers, ciliate protozoa Tetrahymena thermophila as consumers and bacteria Escherichia coli as decomposers. At 1.1 Gy day(-1), no effects were observed. At 5.1 Gy day(-1), cell densities of E. coli showed a tendency to be lower than those of controls. At 9.7 and 24.7 Gy day(-1), population decrease was observed in E. coli. E. gracilis and T. thermophila died out after temporal population decrease and subsequent population increase in T. thermophila. It is likely that this temporal population increase was an indirect effect due to interspecies interactions. Effect dose rates of γ-rays were compared with effect concentrations of some metals using the radiochemoecological conceptual model and the effect index for microcosm. Comparison of these community-level effects data with environmental exposure data suggests that ionising radiation, gadolinium and dysprosium have low risks to affect aquatic microbial communities while manganese, nickel and copper have considerable risks. Effects of chronic irradiation were smaller than those of acute irradiation, and an acute to chronic ratio was calculated to be 28 by dividing an acute dose by chronic daily dose rate at which the effect index was 10%. This ratio would be useful for community-level extrapolation from acute to chronic radiation effects. Copyright © 2011 Elsevier Ltd. All rights reserved.

Screening for unicellular algae as possible bioassay organisms for monitoring marine water samples.

PubMed

Millán de Kuhn, Rosmary; Streb, Christine; Breiter, Roman; Richter, Peter; Neesse, Thomas; Häder, Donat-Peter

2006-08-01

ECOTOX is an automatic early warning system to monitor potential pollution of freshwater, municipal or industrial waste waters or aquatic ecosystems. It is based on a real time image analysis of the motility and orientation parameters of the unicellular, photosynthetic flagellate Euglena gracilis. In order to widen the use of the device to marine habitats and saline waters nine marine flagellates were evaluated as putative bioassay organisms, viz. Dunaliella salina, Dunaliella viridis, Dunaliella bardawil, Prorocentrum minimum Kattegat, P. minimum Lissabon, Tetraselmis suecica, Heterocapsa triquetra, Gyrodinium dorsum and Cryptomonas maculata. Because of their slow growth the last three strains were excluded from further evaluation. Selection criteria were ease of culture, density of cell suspension, stability of motility and gravitactic orientation. The sensitivity toward toxins was tested using copper(II) ions. The instrument allows the user to automatically determine effect-concentration (EC) curves from which the EC(50) values can be calculated. For the interpretation of the EC curves a sigmoid logistic model was proposed which proved to be satisfactory for all tested strains. The inhibition of the motility was considered as the most appropriate movement parameter as an endpoint. The Dunaliella species had the lowest sensitivity to copper with EC(50) values of 220, 198 and 176 mg/L for D. salina, D. bardawil and D. viridis, respectively, followed by T. suecica with an EC(50) value of 40 mg/L. The Prorocentrum species were found to be the most sensitive with an EC(50) value of 13.5 mg/L for P. minimum Lissabon and 7.5 mg/L for P. minimum Kattegat.

Diversity of microflora in the gut and casts of tropical composting earthworms reared on different substrates.

PubMed

Parthasarathi, K; Ranganathan, L S; Anandi, V; Zeyer, Josef

2007-01-01

The diversity of fungi, bacteria, yeast, actinomycetes and protozoa were analysed in the gut and casts of Eudrilus eugeniae, Lampito mauritii, Eisenia fetida and Perionyx excavatus, both qualitatively and quantitatively as influenced by different feed substrates like clay loam soil, cowdung and pressmud. While actinomycetes (Streptomyces albus, S. somaliensis, Nocardia asteroides, N. caviae and Saccharomonosporia) were not digested by any of these species of worms, protozoa (Amoeba proteus, A. terricola, Paramecium trichium, Euglena viridis, E. orientalis, Vorticella picta and Trichomonas hominis) and yeast (Candida tropicalis, C. krusei C. albicans and Cryptococcus neoformans) were totally digested. Certain species of fungi (Saksenae vasiformis, Mucor plumbeus, Cladosporium carrionii, C. herbacium, Alternaria sp., Cunninghamella echinulata, Mycetia sterila, Syncephalostrum racemosum, Curvalaria lunata, C. geniculata and Geotrichum candidum) and bacteria (Pseudomonas aeruginosa, Bacterium antitratum, Mima polymorpha, Enterobacter aerogenes, E. cloacae, Proteus vulgaris, P. mirabilis, P. rettgeri, Escherichia coli, Staphylococus citreus, Bacillus subtilis, B. cereus, Enterococci and Micrococci) were completely digested. Certain other species were not digested fungi like Aspergillus fumigatus, A. flavus, A. ochraceous, Trichoderma koningii (except by Eeugeniae), Fusarium moniliforme (except by E. eugeniae) and Rhizopus sp., and bacteria like Klebsiella pneumoniae and Morganella morganii) and these were multiplied during the transit of the organic residues through the gut of worms. The microbial proliferation was more in the casts, due to the environment prevailing--rich in nutrient supply and large surface area available for growth and reproduction of the microbes that lead to enhanced microbial activity and humic acid contents in the casts.

Saltatory ontogeny of fishes and sensitive early life stages for ecotoxicology tests.

PubMed

Belanger, Scott E; Balon, Eugene K; Rawlings, Jane M

2010-04-15

Fish display a wide range of developmental ontogenies. These distinctions have taxonomic, evolutionary, and ecological importance in addition to practical implications on the use of fish in aquatic toxicity tests. With respect to animal welfare, vertebrates are afforded protected or non-protected status in the European Union based upon whether they feed endogenously off the yolk or exogenously by procurement and ingestion of food. The concept of saltatory ontogeny suggests development is not gradual but proceeds in leaps separated by a series of stable developmental states. In this context, endogenous/exogenous feeding also distinguishes the developmental phases of embryo (egg), eleutheroembryo (feeding off the yolk sac) and larvae (exogenous feeding) in fish. The recent proposal for the Fish Embryo Test (FET) as an animal alternative to the standard fish acute toxicity test (OECD 203 and equivalent tests) puts a clear focus on the need to identify the non-protected and protected life intervals in test species as well as their sensitivities which coincides with the developmental phases identified in saltatory ontogeny. In this paper we described a method to quantify embryo, eleutheroembryo, and larva phases in Danio rerio, the zebrafish. Danio eleutheroembryos preyed upon 5 different protozoan species (Euglena, Euplotes, Paramecium aurelia, Paramecium bursaria and Paramecium multimicronucleatum) between 24 and 48hr following hatching (85-95% of fish, n=20 per species, 25 degrees C). Based upon these data it is recommended that testing of developing zebrafish embryos should be terminated between 24 and 48hr after hatching in order to be compliant with existing animal welfare legislation within Europe.

Analysis of the nutritional status of algae by Fourier transform infrared chemical imaging

NASA Astrophysics Data System (ADS)

Hirschmugl, Carol J.; Bayarri, Zuheir-El; Bunta, Maria; Holt, Justin B.; Giordano, Mario

2006-09-01

A new non-destructive method to study the nutritional status of algal cells and their environments is demonstrated. This approach allows rapid examination of whole cells without any or little pre-treatment providing a large amount of information on the biochemical composition of cells and growth medium. The method is based on the analysis of a collection of infrared (IR) spectra for individual cells; each spectrum describes the biochemical composition of a portion of a cell; a complete set of spectra is used to reconstruct an image of the entire cell. To obtain spatially resolved information synchrotron radiation was used as a bright IR source. We tested this method on the green flagellate Euglena gracilis; a comparison was conducted between cells grown in nutrient replete conditions (Type 1) and on cells allowed to deplete their medium (Type 2). Complete sets of spectra for individual cells of both types were analyzed with agglomerative hierarchical clustering, leading to distinct clusters representative of the two types of cells. The average spectra for the clusters confirmed the similarities between the clusters and the types of cells. The clustering analysis, therefore, allows the distinction of cells of the same species, but with different nutritional histories. In order to facilitate the application of the method and reduce manipulation (washing), we analyzed the cells in the presence of residual medium. The results obtained showed that even with residual medium the outcome of the clustering analysis is reliable. Our results demonstrate the applicability FTIR microspectroscopy for ecological and ecophysiological studies.

Nanophase Iron Oxides as an Ultraviolet Sunscreen for Ancient Photosynthetic Microbes: A Possible Link Between Early Organisms, Banded-Iron Formations, and the Oxygenation of the Atmosphere

NASA Technical Reports Server (NTRS)

Bishop, Janice L.; Rothschild, Lynn J.; Rothschild, Lynn J.; Rogoff, Dana A.

2006-01-01

We propose that nanophase iron oxide-bearing materials provided important niches for ancient photosynthetic microbes on the early Earth that ultimately led to the oxygenation of the Earth s atmosphere and the formation of iron oxide deposits. Atmospheric oxygen and ozone attenuate UV radiation on the Earth today providing substantial protection for photosynthetic organisms. With ultraviolet radiation fluxes likely to have been even higher on the early Earth than today, accessing solar radiation was particularly risky for early organisms. Yet, we know that photosynthesis arose then and played a critical role in subsequent evolution. Of primary importance was protection at approx.250-290 nm, where peak nucleic acid (approx.260 nm) and protein (approx.280 nm) absorptions occur. Nanophase ferric oxide/oxyhydroxide minerals absorb, and thus block, the lethal UV radiation, while transmitting light through much of the visible and near-infrared regions of interest to photosynthesis (400 to 1100 nm). Further, they were available in early environments, and are synthesized by many organisms. Based on ferric oxide/oxyhydroxide spectral properties, likely geologic processes, and the results of experiments with the photosynthetic organisms, Euglena sp. and Chlumydomonus reinhardtii, we propose a scenario where photosynthesis, and ultimately the oxygenation of the atmosphere, depended on the protection of early microbes by nanophase ferric oxides/oxyhydroxides. The results of this study are also applicable to other potentially habitable iron-bearing planetary bodies because of the evolutionary pressure to utilize solar radiation when available as an energy source.

Temporal variation in plankton assemblages and physicochemistry of Devils Lake, North Dakota

USGS Publications Warehouse

Leland, H.V.; Berkas, W.R.

1998-01-01

Seasonal and annual variation in biomass and structure of algal assemblages of hyposaline Devils Lake were examined in relation to turbidity, ambient concentrations of major ions, trace elements and nutrients, and the standing crop of herbivores. Lake level declined during the early years of study, but rose markedly in subsequent years as historically large volumes of water flowed into this hydrologically-closed basin. Winter algal assemblages were dominated (in biomass) most years by small, non-motile chlorophytes ( Choricystis minor, Kirchneriella lunaris or Dunaliella sp.), or Euglena sp. in the most saline sub-basin. Spring assemblages were dominated by diatoms (Stephanodiscus cf. minutulus, Surirella peisonis, Cyclotella meneghiniana and Entomoneis paludosa were especially prominent) or chlorophytes ( C. minor) until the lake level rose. C. minor abundances then declined in spring assemblages and diatoms ( Stephanodiscus cf. agassizensis and S. niagarae; E. paludosa in the more saline sub-basins) dominated. The potential for nitrogen-deficient conditions for phytoplankton growth was evidenced most summers and early autumns by consistently high concentrations of reactive-P relative to inorganic-N and blooms of the N-fixing cyanophyte Aphanizomenon flos-aquae; Microcystis aeruginosa typically was a co-dominant (>30% of biomass) in these assemblages. Pulses of diatoms ( S. cf. agassizensis and C. meneghiniana) occurred in summers following unusually prolonged periods of calm weather or large water inflows. Physical (irradiance, turbulence) and chemical (major nutrients) variables were the primary factors associated with phytoplankton growth. Transparency and major nutrient concentrations accounted for more of the annual variation in phytoplankton structure than did salinity. Seasonal abundance patterns of the dominant zooplankton (the copepod Diaptomus sicilis; the cladocerans Ceriodaphnia quadrangula, Chydorus sphaericus, Daphnia pulex and Diaphanosoma birgei; and the rotifers Brachionus spp., Filinia longiseta, Keratella cochlearis and K. quadrata) also indicated variation in algal populations related to grazing.

The evolution of glutathione metabolism in phototrophic microorganisms

NASA Technical Reports Server (NTRS)

Fahey, R. C.; Buschbacher, R. M.; Newton, G. L.

1987-01-01

Of the many roles ascribed to glutathione (GSH) the one most clearly established is its role in the protection of higher eucaryotes against oxygen toxicity through destruction of thiol-reactive oxygen byproducts. If this is the primary function of GSH then GSH metabolism should have evolved during or after the evolution of oxygenic photosynthesis. That many bacteria do not produce GSH is consistent with this view. In the present study we have examined the low-molecular-weight thiol composition of a variety of phototrophic microorganisms to ascertain how evolution of GSH production is related to evolution of oxygenic photosynthesis. Cells were extracted in the presence of monobromobimane (mBBr) to convert thiols to fluorescent derivatives, which were analyzed by high-pressure liquid chromatography. Significant levels of GSH were not found in the green bacteria (Chlorobium thiosulfatophilum and Chloroflexus aurantiacus). Substantial levels of GSH were present in the purple bacteria (Chromatium vinosum, Rhodospirillum rubrum, Rhodobacter sphaeroides, and Rhodocyclus gelatinosa), the cyanobacteria [Anacystis nidulans, Microcoleus chthonoplastes S.G., Nostoc muscorum, Oscillatoria amphigranulata, Oscillatoria limnetica, Oscillatoria sp. (Stinky Spring, Utah), Oscillatoria terebriformis, Plectonema boryanum, and Synechococcus lividus], and eucaryotic algae (Chlorella pyrenoidsa, Chlorella vulgaris, Euglena gracilis, Scenedesmus obliquus, and Chlamydomonas reinhardtii). Other thiols measured included cysteine, gamma-glutamylcysteine, thiosulfate, coenzyme A, and sulfide; several unidentified thiols were also detected. Many of the organisms examined also exhibited a marked ability to reduce mBBr to syn-(methyl,methyl)bimane, an ability that was quenched by treatment with 2-pyridyl disulfide or 5,5'-bisdithio-(2-nitrobenzoic acid) prior to reaction with mBBr. These observations indicate the presence of a reducing system capable of electron transfer to mBBr and reduction of reactive disulfides. The distribution of GSH in phototrophic eubacteria indicates that GSH synthesis evolved at or around the time that oxygenic photosynthesis evolved.

High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy.

PubMed

Guo, Baoshan; Lei, Cheng; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

2017-05-01

The development of reliable, sustainable, and economical sources of alternative fuels to petroleum is required to tackle the global energy crisis. One such alternative is microalgal biofuel, which is expected to play a key role in reducing the detrimental effects of global warming as microalgae absorb atmospheric CO 2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid amounts and fail to characterize a diverse population of microalgal cells with single-cell resolution in a non-invasive and interference-free manner. Here high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy was demonstrated. In particular, Euglena gracilis, an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement), within lipid droplets was investigated. The optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch quantitative phase microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase maps of every single cell at a high throughput of 10,000 cells/s, enabling accurate cell classification without the need for fluorescent staining. Specifically, the dataset was used to characterize heterogeneous populations of E. gracilis cells under two different culture conditions (nitrogen-sufficient and nitrogen-deficient) and achieve the cell classification with an error rate of only 2.15%. The method holds promise as an effective analytical tool for microalgae-based biofuel production. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Localizing Proteins in Fixed Giardia lamblia and Live Cultured Mammalian Cells by Confocal Fluorescence Microscopy.

PubMed

Nyindodo-Ogari, Lilian; Schwartzbach, Steven D; Skalli, Omar; Estraño, Carlos E

2016-01-01

Confocal fluorescence microscopy and electron microscopy (EM) are complementary methods for studying the intracellular localization of proteins. Confocal fluorescence microscopy provides a rapid and technically simple method to identify the organelle in which a protein localizes but only EM can identify the suborganellular compartment in which that protein is present. Confocal fluorescence microscopy, however, can provide information not obtainable by EM but required to understand the dynamics and interactions of specific proteins. In addition, confocal fluorescence microscopy of cells transfected with a construct encoding a protein of interest fused to a fluorescent protein tag allows live cell studies of the subcellular localization of that protein and the monitoring in real time of its trafficking. Immunostaining methods for confocal fluorescence microscopy are also faster and less involved than those for EM allowing rapid optimization of the antibody dilution needed and a determination of whether protein antigenicity is maintained under fixation conditions used for EM immunogold labeling. This chapter details a method to determine by confocal fluorescence microscopy the intracellular localization of a protein by transfecting the organism of interest, in this case Giardia lamblia, with the cDNA encoding the protein of interest and then processing these organisms for double label immunofluorescence staining after chemical fixation. Also presented is a method to identify the organelle targeting information in the presequence of a precursor protein, in this case the presequence of the precursor to the Euglena light harvesting chlorophyll a/b binding protein of photosystem II precursor (pLHCPII), using live cell imaging of mammalian COS7 cells transiently transfected with a plasmid encoding a pLHCPII presequence fluorescent protein fusion and stained with organelle-specific fluorescent dyes.

Meneco, a Topology-Based Gap-Filling Tool Applicable to Degraded Genome-Wide Metabolic Networks

PubMed Central

Prigent, Sylvain; Frioux, Clémence; Dittami, Simon M.; Larhlimi, Abdelhalim; Collet, Guillaume; Gutknecht, Fabien; Got, Jeanne; Eveillard, Damien; Bourdon, Jérémie; Plewniak, Frédéric; Tonon, Thierry; Siegel, Anne

2017-01-01

Increasing amounts of sequence data are becoming available for a wide range of non-model organisms. Investigating and modelling the metabolic behaviour of those organisms is highly relevant to understand their biology and ecology. As sequences are often incomplete and poorly annotated, draft networks of their metabolism largely suffer from incompleteness. Appropriate gap-filling methods to identify and add missing reactions are therefore required to address this issue. However, current tools rely on phenotypic or taxonomic information, or are very sensitive to the stoichiometric balance of metabolic reactions, especially concerning the co-factors. This type of information is often not available or at least prone to errors for newly-explored organisms. Here we introduce Meneco, a tool dedicated to the topological gap-filling of genome-scale draft metabolic networks. Meneco reformulates gap-filling as a qualitative combinatorial optimization problem, omitting constraints raised by the stoichiometry of a metabolic network considered in other methods, and solves this problem using Answer Set Programming. Run on several artificial test sets gathering 10,800 degraded Escherichia coli networks Meneco was able to efficiently identify essential reactions missing in networks at high degradation rates, outperforming the stoichiometry-based tools in scalability. To demonstrate the utility of Meneco we applied it to two case studies. Its application to recent metabolic networks reconstructed for the brown algal model Ectocarpus siliculosus and an associated bacterium Candidatus Phaeomarinobacter ectocarpi revealed several candidate metabolic pathways for algal-bacterial interactions. Then Meneco was used to reconstruct, from transcriptomic and metabolomic data, the first metabolic network for the microalga Euglena mutabilis. These two case studies show that Meneco is a versatile tool to complete draft genome-scale metabolic networks produced from heterogeneous data, and to suggest relevant reactions that explain the metabolic capacity of a biological system. PMID:28129330

Selective control of toxic Microcystis water blooms using lysine and malonic acid: an enclosure experiment.

PubMed

Kaya, Kunimitsu; Liu, Yong-Ding; Shen, Yin-Wu; Xiao, Bang-Ding; Sano, Tomoharu

2005-04-01

Three enclosures (10 x 10 x 1.5-1.3 m in depth) were set beside Dianch Lake, Kunming, People's Republic of China, for the period from July 28 to August 26, 2002. The enclosures were filled with cyanobacterial (Microcystis aeruginosa) water bloom-containing lake water. Lake sediment that contained macrophytes and water chestnut seeds was spread over the entire bottom of each enclosure. Initially, 10 g/m(2) of lysine was sprayed in Enclosure B, and 10 g/m(2) each of lysine and malonic acid were sprayed together in Enclosure C. Enclosure A remained untreated and was used as a control. The concentrations of lysine, malonic acid, chlorophyll a, and microcystin as well as the cell numbers of phytoplankton such as cyanobacteria, diatom, and euglena were monitored. On day 1 of the treatment, formation of cyanobacterial blooms almost ceased in Enclosures B and C, although Microcystis cells in the control still formed blooms. On day 7 Microcystis cells in Enclosure B that had been treated with lysine started growing again, whereas growth was not observed in Microcystis cells in Enclosure C, which had been treated with lysine and malonic acid. On day 28 the surface of Enclosure B was covered with water chestnut (Trapa spp.) and the Microcystis blooms again increased. In contrast, growth of macrophytes (Myriophllum spicatum and Potamogeton crispus) was observed in Enclosure C; however, no cyanobacterial blooms were observed. Lysine and malonic acid had completely decomposed. The microcystin concentration on day 28 decreased to 25% of the initial value, and the pH shifted from the initial value of 9.2 to 7.8. We concluded that combined treatment with lysine and malonic acid selectively controlled toxic Microcystis water blooms and induced the growth of macrophytes.

Algae in relation to mine water

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bennett, H.D.

1969-01-01

An annual cycle of bimonthly collections was made from 17 stations located on creeks, rivers, and ponds receiving acid mine drainage in order to obtain information on the species of algae that are tolerant to these waters. Also data were obtained to determine the relative importance of some of the major chemical factors of the water to ecology of the algae. Nitrate, phosphate, and calcium did not appear to be limiting or modifying. There was a lack of correlation between chemical factors except for total acidity, iron and pH. A range for the latter three characterized each of the habitatsmore » studied. Total acidity and the associated factors (iron and pH) appeared to have the controlling influence on the algal population in the more highly acid streams. The total number of genera and species as observed in a living condition in mine polluted water, compared favorably with numbers reported from unpolluted waters. Nearly half of the total species observed were found in the more highly acid creeks, as well as in the other habitats (less acid creeks, rivers and ponds). Algae characteristic of mine polluted water were found to be those common to a range of habitats, with the reduction in numbers of species at higher acidities and at lower pH values, being primariy in those that are less common to a range of habitats. The range of total acidity and pH values at a particular site or stream reach can be characterized by a range in the number of species and their abundance - an increase or decrease in abundance being dependent on the algal species. Some species such as Euglena mutabilis Sch., Eunotia tenella (grun) Gleve, and Pinnularia braunii (grun) Cleve, are most abundant in mine polluted water.« less

Crystal structure of homoisocitrate dehydrogenase from Schizosaccharomyces pombe

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bulfer, Stacie L.; Hendershot, Jenna M.; Trievel, Raymond C.

Lysine biosynthesis in fungi, euglena, and certain archaebacteria occurs through the {alpha}-aminoadipate pathway. Enzymes in the first steps of this pathway have been proposed as potential targets for the development of antifungal therapies, as they are absent in animals but are conserved in several pathogenic fungi species, including Candida, Cryptococcus, and Aspergillus. One potential antifungal target in the {alpha}-aminoadipate pathway is the third enzyme in the pathway, homoisocitrate dehydrogenase (HICDH), which catalyzes the divalent metal-dependent conversion of homoisocitrate to 2-oxoadipate (2-OA) using nicotinamide adenine dinucleotide (NAD{sup +}) as a cofactor. HICDH belogns to a family of {beta}-hydroxyacid oxidative decarboxylases thatmore » includes malate dehydrogenase, tartrate dehydrogenase, 6-phosphogluconate dehydrogenase, isocitrate dehydrogenase (ICDH), and 3-isopropylmalte dehydrogenase (IPMDH). ICDH and IPMDH are well-characterized enzymes that catalyze the decarboxylation of isocitrate to yield 2-oxoglutarate (2-OG) in the citric acid cycle and the conversion of 3-isopropylmalate to 2-oxoisovalerate in the leucine biosynthetic pathway, respectively. Recent structural and biochemical studies of HICDH reveal that this enzyme shares sequence, structural, and mechanistic homology with ICDH and IPMDH. To date, the only published structures of HICDH are from the archaebacteria Thermus thermophilus (TtHICDH). Fungal HICDHs diverge from TtHICDH in several aspects, including their thermal stability, oligomerization state, and substrate specificity, thus warranting further characterization. To gain insights into these differences, they determined crystal structures of a fungal Schizosaccharomyces pombe HICDH (SpHICDH) as an apoenzyme and as a binary complex with additive tripeptide glycyl-glycyl-glycine (GGG) to 1.55 {angstrom} and 1.85 {angstrom} resolution, respectively. Finally, a comparison of the SpHICDH and TtHICDH structures reveal differences in their active sites that help explain the variations in their respective substrate specificities.« less

Contribution of eukaryotic microbial communities to the formation of Fe-rich accretions in an extreme acidic environment

NASA Astrophysics Data System (ADS)

Rodrigues, L.; Valente, T.; Correia, A.; Alves, A.; Foing, B.; Davies, G. R.

2012-04-01

In the acid mine drainage of Valdarcas, northern Portugal, Fe-rich tubular and spherical macroaccretions are directly associated with the presence of eukaryotic microorganisms. This raises the question whether they are biogenically-derived or the result of an abiotic process mediated by microeukaryotic phototrophs. The drainage water at Valdarcas is characterized by very low pH values (pH<3.5), high metal solubility and presence of iron colloids. Mineralogical analysis (XRD and SEM) of the precipitates indicates a mixture of goethite, schwertmannite and jarosite. Euglenophyta and Chlorophyta acidophilic algal were previously identified in this site. The spatial distribution of Euglena mutabilis indicated that it has a preference to grow up on schwertmannite-rich precipitates. Field observations demonstrate the existence of oxygenated microenvironments created by algal activity suggesting that algae influence iron minerals precipitation, especially schwertmannite. The mineral-microorganism interactions are relevant to understanding this unique and extreme environment. Further investigations regarding the mineralogical and chemical characterization of these deposits, and the identification of microorganisms involved in the process could be helpful to enhance our knowledge of past Fe formations throughout Earth's primordial environment. It is expectable that this information will contribute to establish a framework for recognition of biosignatures on other planets and extraterrestrial bodies. In this study, results on the chemical and mineralogical composition of the structures are presented. The biological context is characterised based on observations made by optical microscopy complemented with molecular data on the microbial communities obtained by culture independent methods. The results are discussed within the context of two models: the studied Fe-rich stromatolites are microeukaryotic-mediated as described by previous workers from similar environments or are the consequence of inorganic precipitation of reduced iron species (Fe(III)) due to the oxygen generated by the photosynthesis?

Meneco, a Topology-Based Gap-Filling Tool Applicable to Degraded Genome-Wide Metabolic Networks.

PubMed

Prigent, Sylvain; Frioux, Clémence; Dittami, Simon M; Thiele, Sven; Larhlimi, Abdelhalim; Collet, Guillaume; Gutknecht, Fabien; Got, Jeanne; Eveillard, Damien; Bourdon, Jérémie; Plewniak, Frédéric; Tonon, Thierry; Siegel, Anne

2017-01-01

Increasing amounts of sequence data are becoming available for a wide range of non-model organisms. Investigating and modelling the metabolic behaviour of those organisms is highly relevant to understand their biology and ecology. As sequences are often incomplete and poorly annotated, draft networks of their metabolism largely suffer from incompleteness. Appropriate gap-filling methods to identify and add missing reactions are therefore required to address this issue. However, current tools rely on phenotypic or taxonomic information, or are very sensitive to the stoichiometric balance of metabolic reactions, especially concerning the co-factors. This type of information is often not available or at least prone to errors for newly-explored organisms. Here we introduce Meneco, a tool dedicated to the topological gap-filling of genome-scale draft metabolic networks. Meneco reformulates gap-filling as a qualitative combinatorial optimization problem, omitting constraints raised by the stoichiometry of a metabolic network considered in other methods, and solves this problem using Answer Set Programming. Run on several artificial test sets gathering 10,800 degraded Escherichia coli networks Meneco was able to efficiently identify essential reactions missing in networks at high degradation rates, outperforming the stoichiometry-based tools in scalability. To demonstrate the utility of Meneco we applied it to two case studies. Its application to recent metabolic networks reconstructed for the brown algal model Ectocarpus siliculosus and an associated bacterium Candidatus Phaeomarinobacter ectocarpi revealed several candidate metabolic pathways for algal-bacterial interactions. Then Meneco was used to reconstruct, from transcriptomic and metabolomic data, the first metabolic network for the microalga Euglena mutabilis. These two case studies show that Meneco is a versatile tool to complete draft genome-scale metabolic networks produced from heterogeneous data, and to suggest relevant reactions that explain the metabolic capacity of a biological system.

Anaerobic Coculture of Microalgae with Thermosipho globiformans and Methanocaldococcus jannaschii at 68°C Enhances Generation of n-Alkane-Rich Biofuels after Pyrolysis

PubMed Central

Matsuyama, Shigeru; Igarashi, Kensuke; Utsumi, Motoo; Shiraiwa, Yoshihiro; Kuwabara, Tomohiko

2013-01-01

We tested different alga-bacterium-archaeon consortia to investigate the production of oil-like mixtures, expecting that n-alkane-rich biofuels might be synthesized after pyrolysis. Thermosipho globiformans and Methanocaldococcus jannaschii were cocultured at 68°C with microalgae for 9 days under two anaerobic conditions, followed by pyrolysis at 300°C for 4 days. Arthrospira platensis (Cyanobacteria), Dunaliella tertiolecta (Chlorophyta), Emiliania huxleyi (Haptophyta), and Euglena gracilis (Euglenophyta) served as microalgal raw materials. D. tertiolecta, E. huxleyi, and E. gracilis cocultured with the bacterium and archaeon inhibited their growth and CH4 production. E. huxleyi had the strongest inhibitory effect. Biofuel generation was enhanced by reducing impurities containing alkanenitriles during pyrolysis. The composition and amounts of n-alkanes produced by pyrolysis were closely related to the lipid contents and composition of the microalgae. Pyrolysis of A. platensis and D. tertiolecta containing mainly phospholipids and glycolipids generated short-carbon-chain n-alkanes (n-tridecane to n-nonadecane) and considerable amounts of isoprenoids. E. gracilis also produced mainly short n-alkanes. In contrast, E. huxleyi containing long-chain (31 and 33 carbon atoms) alkenes and very long-chain (37 to 39 carbon atoms) alkenones, in addition to phospholipids and glycolipids, generated a high yield of n-alkanes of various lengths (n-tridecane to n-pentatriacontane). The gas chromatography-mass spectrometry (GC-MS) profiles of these n-alkanes were similar to those of native petroleum crude oils despite containing a considerable amount of n-hentriacontane. The ratio of phytane to n-octadecane was also similar to that of native crude oils. PMID:23183975

Gravitaxis and graviperception in flagellates.

PubMed

Hader, D-P; Lebert, M; Richter, P; Ntefidou, M

2003-01-01

There is strong evidence that gravitactic orientation in flagellates and ciliates is mediated by an active physiological gravireceptor rather than by passive alignment of the cells in the water column. In flagellates the threshold for graviorientation was found to be at 0.12 x g on a slow rotating centrifuge during the IML-2 mission on the Shuttle Columbia and a subsequent parabolic rocket flight (TEXUS). During the IML-2 mission no adaptation to microgravity was observed over the duration of the space flight, while gravitaxis was lost in a terrestrial closed environmental system over the period of almost two years. Sedimenting statoliths are not likely to be involved in graviperception because of the small size of the cells and their rotation around the longitudinal axis during forward locomotion. Instead the whole cytoplasmic content of the cell, being heavier than the surrounding aqueous medium (1.05 g/ml), exerts a pressure on the lower membrane. This force activates stretch-sensitive calcium specific ion channels which can be inhibited by the addition of gadolinium which therefore abolishes gravitaxis. The channels seem to mainly allow calcium ions to pass since gravitaxis is blocked by the addition of the calcium ionophore A23187 and by vanadate which blocks the Ca-ATPase in the cytoplasmic membrane. Recently, a gene for a mechanosensitive channel, originally sequenced for Saccharomyces, was identified in Euglena by PCR. The increase in intracellular free calcium during reorientation can be visualized by the fluorophore Calcium Crimson using laser excitation and image intensification. This result was confirmed during recent parabolic flights. The gated calcium changes the membrane potential across the membrane which may be the trigger for the reorientation of the flagellum. cAMP plays a role as a secondary messenger. Photosynthetic flagellates are suitable candidates for life support systems since they absorb CO2 and produce oxygen. Preliminary experiments are discussed. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

A Mach-Zender Holographic Microscope for Quantifying Bacterial Motility

NASA Astrophysics Data System (ADS)

Niraula, B.; Nadeau, J. L.; Serabyn, E.; Wallace, J. K.; Liewer, K.; Kuhn, J.; Graff, E.; Lindensmith, C.

2014-12-01

New microscopic techniques have revolutionized cell biology over the past two decades. However, there are still biological processes whose details elude us, especially those involving motility: e.g. feeding behavior of microorganisms in the ocean, or migration of cancer cells to form metastases. Imaging prokaryotes, which range in size from several hundred nm to a few microns, is especially challenging. An emerging technique to address these issues is Digital Holographic Microscopy (DHM). DHM is an imaging technique that uses the interference of light to record and reproduce three-dimensional magnified images of objects. This approach has several advantages over ordinary brightfield microscopy for fieldwork: a larger depth of field, hands-off operation, robustness regarding environmental conditions, and large sampling volumes with quantitative 3D records of motility behavior. Despite these promising features, real-time DHM was thought to be impractical for technological and computational reasons until recently, and there has so far been very limited application of DHM to biology. Most existing instruments are limited in performance by their particular (e.g. in-line, lens-less, phase-shifting) approach to holography. These limitations can be mitigated with an off-axis dual-path configuration. Here we describe the design and implementation of a design for a Mach-Zehnder-type holographic microscope with diffraction-limited lateral resolution, with intended applications in environmental microbiology. We have achieved sub-micron resolution and three-dimensional tracking of prokaryotic and eukaryotic test strains designed to represent different modes and speeds of microbial motility. Prokaryotes are Escherichia coli, Vibrio alginolyticus, and Bacillus subtilis. Each shows a characteristic motility pattern, as we illustrate in holographic videos in sample chambers 0.6 mm in depth. The ability to establish gradients of attractants with bacterial taxis towards the attractant is also established. The eukaryotic strains are Euglena gracilis, which demonstrates both phototaxis and geotaxis, and Paramecium micromultinucleatum. The challenges of optimizing resolution vs. field of view, and of handling the large volumes of data generated during holographic imaging, are discussed.

[Trophic ecology in tadpoles of Rhinella arenarum (Anura: Bufonidae) in agroecosystems and their possible implications for conservation].

PubMed

Bionda, Clarisa; Gari, Noemi; Luque, Elisa; Salas, Nancy; Lajmanovich, Rafael; Martino, Adolfo

2012-06-01

The progress of the agriculture border has led an important loss of natural habitats, with significant consequences for biodiversity. In this sense, the studies in anuran amphibian tadpoles inhabiting these environments are relevant, because the larval stage is a phase of population regulation. The aim of this study was to analyze the body condition and diet in Rhinella arenarum, tadpoles, an anuran species widely distributed in South America and that inhabit agroecosystems. Three sites were sampled, two agroecosystems with different alteration degrees (C1 and C2) and an uncultured (SM) third place. The captured tadpoles were anesthetized, fixed and preserved in formaldehyde (10%). Subsequently, body measurements were made and the complete intestine was removed and analyzed for food items under a binocular microscope. The diet in R. arenarum tadpoles has a dominance of algae Bacillariophyceae, followed by Cyanophyceae. In particular, the class Bacillariophyceae, due to the presence of the genus Navicula, Nitzschia, Gomphonema and Hantzschia, was important in the diet of the anurans in those agroecosystems. Class Cyanophyceae, mainly represented by genus Oscillatoria and Euglenophyceae represented by Euglena and Strombomonas, were predominant in the diet of the anurans in SM. Some differences in the total items consumed by tadpole were observed between the studied sites. Tadpoles that inhabit the modified sites (C1 and C2) recorded a significantly smaller amount of food. Moreover, the tadpoles that inhabit these sites showed a lower body condition. The presence of certain algae associated with eutrophic environments, could indicate some pollution in agroecosystems (C1 and C2). Food resources would be lesser in places with strong agricultural activity, possibly with a greater degree of eutrophication. A smaller food amount could have consequences at population level for the short and long time terms, because of its impact on individual growth. Larval diet is suggested as a potential bioindicator of environmental health for these areas.

Photoreception in Phytoplankton.

PubMed

Colley, Nansi Jo; Nilsson, Dan-Eric

2016-11-01

In many species of phytoplankton, simple photoreceptors monitor ambient lighting. Photoreceptors provide a number of selective advantages including the ability to assess the time of day for circadian rhythms, seasonal changes, and the detection of excessive light intensities and harmful UV light. Photoreceptors also serve as depth gauges in the water column for behaviors such as diurnal vertical migration. Photoreceptors can be organized together with screening pigment into visible eyespots. In a wide variety of motile phytoplankton, including Chlamydomonas, Volvox, Euglena, and Kryptoperidinium, eyespots are light-sensitive organelles residing within the cell. Eyespots are composed of photoreceptor proteins and typically red to orange carotenoid screening pigments. This association of photosensory pigment with screening pigment allows for detection of light directionality, needed for light-guided behaviors such as positive and negative phototaxis. In Chlamydomonas, the eyespot is located in the chloroplast and Chlamydomonas expresses a number of photosensory pigments including the microbial channelrhodopsins (ChR1 and ChR2). Dinoflagellates are unicellular protists that are ecologically important constituents of the phytoplankton. They display a great deal of diversity in morphology, nutritional modes and symbioses, and can be photosynthetic or heterotrophic, feeding on smaller phytoplankton. Dinoflagellates, such as Kryptoperidinium foliaceum, have eyespots that are used for light-mediated tasks including phototaxis. Dinoflagellates belonging to the family Warnowiaceae have a more elaborate eye. Their eye-organelle, called an ocelloid, is a large, elaborate structure consisting of a focusing lens, highly ordered retinal membranes, and a shield of dark pigment. This complex eye-organelle is similar to multicellular camera eyes, such as our own. Unraveling the molecular makeup, structure and function of dinoflagellate eyes, as well as light-guided behaviors in phytoplankton can inform us about the selective forces that drove evolution in the important steps from light detection to vision. We show here that the evolution from simple photoreception to vision seems to have independently followed identical paths and principles in phytoplankton and animals, significantly strengthening our understanding of this important biological process. © The Author 2016. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

OMEGAHAB-XP a bioregenerative aquatic life support system designed to be used in Bion-M1 long term space flight

NASA Astrophysics Data System (ADS)

Hilbig, Reinhard; Lebert, Michael

The OmegaHab XP Experiment will be based on the OmegaHab system successfully flown in the context of the FOTON M3 mission. OmegaHab XP -a refurbished OmegaHab for a long term mission -is in general assembled from four parts: an algae compartment, a nutrition com-partment for higher plants and crustaceens, a fish compartment and a filter compartment with biodegradant bacterias. The algae compartment (Euglena gracilis; unicellular, photosynthetic flagellate) will be illuminated with photosynthetic active radiation and will produce oxygen. The photosynthetic process also consumes carbon dioxide and if available ammonia. In addi-tion, nitrate will be taken up by the algae and by this means removed from the system. Via a gas-permeable membrane (gas/ion exchanger) the produced oxygen will be transported in a separate fish compartment. The metabolism of the fish will produce carbon dioxide and nitro-genic components. These components as well as the carbon dioxide will be transported back in the algae compartment and subsequently used by the algae. The transport of the components is enhanced by a counter flow inside the gas/ion exchanger driven by a pump. In addition, a filter system is installed which removes debris as well as ammonia by means of ammonia metabolizing bacteria. The nutrition compartment with higher plants and the crustaceans (e.g. Hyalella azteca; flown successfully aboard shuttles) builds the basis of this multi-trophic sys-tem. Hyalella azteca can reproduce in an adequate amount to replace external fish nutrition for Oreochromis mossambicus in large parts. The fish compartment is divided into two chambers: a hatchery chamber for larval fishes and an chamber for subadult Oreochromis mossambicus. The system is fully automatic and measures and stores all house-keeping data internally. These house-keeping data include light, temperature, acceleration and oxygen as well as many system related parameters. By means of Peltier-elements the system can be temperature-controlled. Two video cameras (one coupled to a sub-miniaturized microscope, one for observation of the fish) allow to monitor and internally store the behaviour of fish and algae.

Improving Satellite Compatible Microdevices to Study Biology in Space

NASA Technical Reports Server (NTRS)

Kalkus, Trevor; Snyder, Jessica; Paulino-Lima, Ivan; Rothschild, Lynn

2017-01-01

The technology for biology in space lags far behind the gold standard for biological experiments on Earth. To remedy this disparity, the Rothschild lab works on proof of concept, prototyping, and developing of new sensors and devices to further the capabilities of biology research on satellites. One such device is the PowerCell Payload System. One goal for synthetic biology in aiding space travel and colonization is to genetically engineer living cells to produce biochemicals in space. However, such farming in space presupposes bacteria retain their functionality post-launch, bombarded by radiation, and without the 1G of Earth. Our questions is, does a co-culture of cyanobacteria and protein-synthesizing bacteria produce Earth-like yields of target proteins? Is the yield sensitive to variable gravitational forces? To answer these questions, a PowerCell Payload System will spend 1 year aboard the German Aerospace Center's Euglena and Combined Regenerative Organic-food Production In Space (Eu:CROPIS) mission satellite. The PowerCell system is a pair of two 48-well microfluidic cards, each well seeded with bacteria. The system integrates fluidic, thermal, optical, electronic, and control systems to germinate bacteria spores, then measure the protein synthesized for comparison to parallel experiments conducted on the Earth. In developing the PowerCell Payload, we gained insight into the shortcomings of biology experiments on satellites. To address these issues, we have started three new prototyping projects: 1) The development of an extremely stable and radiation resistant cell-free system, allowing for the construction of proteins utilizing only cell components instead of living cells. This can be lyophilized on a substrate, like paper. (2) Using paper as a microfluidic platform that is flexible, stable, cheap, and wicking. The capillary action eliminates the need for pumps, reducing volume, mass, and potential failing points. Electrodes can be printed on the paper to sense for biochemicals. (3) Developing a modular, semi-autonomous microfluidic device that can be easily adapted for a variety of common biological experiments. This versatility will allow for quicker and cheaper experimentation. These improvements to satellite experiment platforms have the potential to radically increase the return from NASA's biological and field studies with reduced development time, mass, and cost with increased robustness data and interpretation.

Intermediate introns in nuclear genes of euglenids - are they a distinct type?

PubMed

Milanowski, Rafał; Gumińska, Natalia; Karnkowska, Anna; Ishikawa, Takao; Zakryś, Bożena

2016-02-29

Nuclear genes of euglenids contain two major types of introns: conventional spliceosomal and nonconventional introns. The latter are characterized by variable non-canonical borders, RNA secondary structure that brings intron ends together, and an unknown mechanism of removal. Some researchers also distinguish intermediate introns, which combine features of both types. They form a stable RNA secondary structure and are classified into two subtypes depending on whether they contain one (intermediate/nonconventional subtype) or both (conventional/intermediate subtype) canonical spliceosomal borders. However, it has been also postulated that most introns classified as intermediate could simply be special cases of conventional or nonconventional introns. Sequences of tubB, hsp90 and gapC genes from six strains of Euglena agilis were obtained. They contain four, six, and two or three introns, respectively (the third intron in the gapC gene is unique for just one strain). Conventional introns were present at three positions: two in the tubB gene (at one position conventional/intermediate introns were also found) and one in the gapC gene. Nonconventional introns are present at ten positions: two in the tubB gene (at one position intermediate/nonconventional introns were also found), six in hsp90 (at four positions intermediate/nonconventional introns were also found), and two in the gapC gene. Sequence and RNA secondary structure analyses of nonconventional introns confirmed that their most strongly conserved elements are base pairing nucleotides at positions +4, +5 and +6/ -8, -7 and -6 (in most introns CAG/CTG nucleotides were observed). It was also confirmed that the presence of the 5' GT/C end in intermediate/nonconventional introns is not the result of kinship with conventional introns, but is due to evolutionary pressure to preserve the purine at the 5' end. However, an example of a nonconventional intron with GC-AG ends was shown, suggesting the possibility of intron type conversion between nonconventional and conventional. Furthermore, an analysis of conventional introns revealed that the ability to form a stable RNA secondary structure by some introns is probably not a result of their relationship with nonconventional introns. It was also shown that acquisition of new nonconventional introns is an ongoing process and can be observed at the level of a single species. In the recently acquired intron in the gapC gene an extended direct repeats at the intron-exon junctions are present, suggesting that double-strand break repair process could be the source of new nonconventional introns.

SEMIANNUAL PROGRESS REPORT ON RADIOBIOLOGY FOR THE PERIOD ENDING DECEMBER 31, 1962

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

1963-10-31

Biochemistry: Progress is reported in studies on alterations in brain lipids with aging and irradiation; investigations on lipids of the starfish, Pisaster ochraceus; the constituents of the leaf cuticle wax of the ornamental rubber plant, Ficus elastica; interactions of blood serum with synthetic polypeptides; and investigations on the source of energy for single beating heart cells. Radiobiology. Progress is reported in studies on the effects of gamma radiation on aqueous solutions of pyrimidine compounds and cysteine, tracer studies of phosphate and sulfur metabolism in chlorella, the morphology and physiology of mitochondria, and photosynthesis and respiration in Euglena gracilis. Pharmacology andmore » Toxicology: The radioprotective activity of 204 amines and amine oxide compounds was investigated and the relation between structure and protective activity is discussed. Progress is reported in studies on the effect of psychotropic drugs on the kangaroo rat and the effect of irradiation on intestinal absorption of nicotinamide by rats. Nuclear Radiology Applications of radioisotope tracer methods in the diagnosis and management of liver and kidney diseases are reported. Progress is reported in the photoscanning of the stomach, heart, liver, and spleen following intravenous injection of inorganic radioiodine or of colloidal aggregates of human serum albumin labeled with I/sup 131/; studies on the antigenicity of colloidal aggregates; animal studies on renal artery occlusion; and studies on the retention of I/sup 131/ labeled human serum albumin and the absorption of Ca/sup 47/ in man. The effects of pretreatment with aminoethylisothiuronium or 5- hydroxytryptamine on survival of irradiated rats were measured. A total of 24 newborn infants was examined in a whole-body counter for radioiodine contamination. No radioiodine or any other radionuclides, except K/sup 40/, was observed. Medicine: Data are summarized from studies on the role of desoxyribonucleic acid and the thymus in leukemogenesis in mice, the development of radioinduced leukemia in mice protected with bone marrow, spleen, and thymus cells frora a strain of mice with a high incidence of leukemia, and the oncogenic properties of desoxyribonucleic acid isolated from parotid gland tumors in mice. Biophysics: Progress is reported in studies on lymphopenia in mice induced by tail irradiation. Environmental Radiation: Data are summarized from a series of studies on the ecological effects of environmental radiation due to fallout. Results are included from studies on the effects of a nuclear detonation on desert vegetation and animal populations. (C.H.)« less

Carrier Dynamics and Application of the Phase Coherent Photorefractive Effect in ZnSe Quantum Wells

NASA Astrophysics Data System (ADS)

Dongol, Amit

The intensity dependent diffraction efficiency of a phase coherent photorefractive (PCP) ZnSe quantum well (QW) is investigated at 80 K in a two-beam four-wave mixing (FWM) configuration using 100 fs laser pulses with a repetition rate of 80 MHz. The observed diffraction efficiencies of the first and second-order diffracted beam are on the order of 10-3 and 10-5, respectively, revealing nearly no intensity dependence. The first-order diffraction is caused by the PCP effect where the probe-pulse is diffracted due to a long-living incoherent electron density grating in the QW. The second-order diffraction is created by a combination of diffraction processes. For negative probe-pulse delay, the exciton polarization is diffracted at the electron grating twice by a cascade effect. For positive delay, the diffracted signal is modified by the destructive interference with a chi(5) generated signal due to a dynamical screening effect. Model calculations of the signal traces based on the optical Bloch equations considering inhomogeneous broadening of exciton energies are in good agreement with the experimental data. To study the carrier dynamics responsible for the occurrence of the PCP effect, threebeam FWM experiments are carried out. The non-collinear wave-vectors k1 , k2 and k3 at central wavelength of 441 nm (~2.81 eV) were resonantly tuned to the heavy-hole exciton transition energy at 20 K. In the FWM experiment the time coincident strong pump pulses k1 and k2 create both an exciton density grating in the QW and an electron-hole pair grating in the GaAs while the delayed weak pulse k3 simultaneously probes the exciton lifetime as well as the electron grating capture time. The model calculations are in good agreement with the experimental results also providing information about the transfer delay of electrons arriving from the substrate to the QW. For negative probe-pulse delay we still observe a diffracted signal due to the long living electron density grating in the QW. The electron grating build-up and decay times are also studied with the modified three-beam FWM set-up. Using an optical shutter for pump pulses k1and k2, the dynamics of the electron grating formation and its decay is continuously probed by a delayed pulse k3. The obtained build-up and decay times are found to depend nearly linearly on the intensity of incident pulses k1 and k2 being on the order of several microseconds at low pump intensities. The PCP effect in ZnSe QW possesses a time-gating capability which can be used for real-time holographic imaging. In this work we demonstrate contrast enhanced real time holographic imaging (CEHI) of floating glass beads and of living unicellular animals (Paramecium and Euglena cells) in aqueous solution. We also demonstrate CEHI of a ~100 im thick wire concealed behind a layer of chicken skin. The results demonstrate the potential of PCP QWs for real-time and depth-resolved imaging of moving micrometer sized biological objects in transparent media or of obscured objects in turbid media.

Biological soil crusts are the main contributor to winter soil respiration in a temperate desert ecosystem of China

NASA Astrophysics Data System (ADS)

He, M. Z.

2012-04-01

Aims Biological soil crusts (BSCs) are a key biotic component of desert ecosystems worldwide. However, most studies carried out to date on carbon (fluxes) in these ecosystems, such as soil respiration (RS), have neglected them. Also, winter RS is reported to be a significant component of annual carbon budget in other ecosystems, however, we have less knowledge about winter RS of BSCs in winter and its contribution to carbon cycle in desert regions. Therefore, the specific objectives of this study were to: (i) quantify the effects of different BSCs types (moss crust, algae crust, physical crust) on the winter RS; (ii) explore relationships of RS against soil temperature and water content for different BSCs, and (iii) assess the relative contribution of BSCs to the annual amount of C released by RS at desert ecosystem level. Methods Site Description The study sites are located at the southeast fringe of the Tengger Desert in the Shapotou region of the Ningxia Hui Autonomous Region [37°32'N and 105°02'E, at 1340 m above mean sea level (a.m.s.l.)], western China. The mean daily temperature in January is -6.9°C , while it is 24.3°C in July. The mean annual precipitation is 186 mm, approximately 80% of which falls between May and September. The annual potential evaporation is 2800 mm. The landscape of the Shapotou region is characterized by large and dense reticulate barchans chains of sand dunes that migrate south-eastward at a velocity of 3-6 m per year. The soil is loose, infertile and mobile and can thus be classified as orthic sierozem and Aeolian sandy soil. Additionally, the soil has a consistent gravimetric water content that ranges from 3 to 4%. The groundwater in the study area is too deep (>60 m) to support large areas of the native vegetation cover; therefore, precipitation is usually the only source of freshwater. The predominant native plants are Hedysarum scoparium Fisch. and Agriophyllum squarrosum Moq., Psammochloa cillosa Bor, which scattered distribute with cover about 1% of the entire study area. Prior to revegetation, straw-checkerboards approximately 1×1 m2 in area were constructed using wheat or rice straw to stabilize the dune surface and allow time for the planted xerophytic shrubs to adapt to the new environment. In 1956, the following 2-year-old xerophytic shrub seedlings were planted within the checkerboard at a density of 16 individuals per 100 m2 and grown without irrigation: Artemisia ordosica Krasch, H. scoparium Fisch, Calligonum mongolicum Turc'z, Caragana microphylla Lam., Caragana korshinskii Kom, Salix gordejevii and Atraphaxis bracteata A.Los. The stabilized area was then expanded to parallel areas in 1964 and 1982 using the same method and species. As a result, the initial stages of change that have occurred at these sites were similar. After more than fifties years succession, the predominant plants are semi-shrubs, shrubs, forbs, and grasses at present and BSCs formed. The common BSCs in the region may be dominated by cyanobacteria, algae, lichens and mosses, or any combination of these organisms. Cyanobacteria species include Microcolous vaginatus Gom., Hydrocoleus violacens Gom., Lyngbya crytoraginatus Schk., Phormidium amblgum Gom., P. autumnale (Ag.) Gom., P. foveolarum (Mont.) Gom. and Phormidium luridum (Kutz) Gom. etc; algal species mainly include Anabaena azotica Ley, Euglena sp., Hantzschia amphioxys var capitata Grum, Oscillatoria obscura Gom., O. pseudogeminate G. Schm. And Scytonema javanicum (Kutz) Bornet Flash etc; lichen species include Collema tenax (Sw.) Ach., Endocarpon pusillum Hedw.; and moss species are dominated by Bryum argenteum Hedw., Didymodon constrictus (Mitt.) Saito., Tortula bidentata Bai Xue Liang and T. desertorum Broth.. Experimental Design and Rs measurements On October 2010, We selected the moss-dominated BSCs at four revegetation sites and natural vegetation sites, in which 3 replicated plots were selected randomly. In each plot, olyvinyl chloride (PVC) collar (lenth 10 cm, internal diameter 10cm ) were inserted 7 cm into the soil. During the establishment of PVC collars, each plot falling within distances less than 2.0 m of an existing replicate was discarded and reallocated to ensure a minimum distance between replicates of 2.0 m. Soil respiration was measured with a LI-6400 and a soil CO2 flux chamber (LI-6400-09). The target value was set close to the ambient CO2 concentration (~380μmolmol_1), and the ΔCO2 value was set as the factory default value (10μmolmol-1) (LiCor 1997). Thin-walled PVC collars were inserted at least one night (12h) before measurement so as to avert CO2 flushing, as recommended by LiCor (1997). Measurements were performed monthly between November 2010 (1months after the collars were inserted to avoid any bias promoted by soil alteration during the placement of collars) and March 2010. Diurnal Rs were measured every two hours from 8:00 to 6:00 of the next day. In parallel to the measurements of soil respiration, Measurements of soil respiration, soil moisture, and soil temperature were taken according to a stratified random design. Important findings (1) The RS was positively correlated to soil temperature in all BSCs, and was significantly influenced by the interactions of soil temperature and water content in moss, lichen and mixed BSCs. (2) The sensitivity of RS to soil temperature at 10cm depth (Q10) ranged from 1.25 in moss-dominated BSCs to 1.63 lichen-dominated BSCs. The Q10 tended to increase with soil temperature until reaching a threshold, and then decline. (3) Soil respiration rates remained low at night time and exhibited as single-peak curve at day time from April to October, and compare to the shifting sand area, a remarkable monthly variation occured in the BSCs dominated area. (4) The winter RS for the moss, lichen, and mixed BSCs averaged 102, 56 and 85 gCm-2 winter time_1, respectively. The winter RS was positively correlated to soil organic carbon (SOC) concentration at O horizon. Our results indicate that winter Rs of BSCs-dominated areas are the main contributor to the total carbon released by soil respiration and, therefore, which we should considered when estimating carbon budgets in desert ecosystems. Key words: Winter Soil Respiration; Biological Soil Crust (BSCs); Q10