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Motile microorganisms such as the green Euglena gracilis use a number of external stimuli to orient in their environment. They respond to light with photophobic responses, photokinesis and phototaxis, all of which can result in accumulations of the organisms in suitable habitats. The light responses operate synergistically with gravitaxis, aerotaxis and other responses. Originally the microscopically obvious stigma was thought to be the photoreceptor, but later the paraxonemal body (PAB, paraflagellar body) has been identified as the light responsive organelle, located in the trailing flagellum inside the reservoir. The stigma can aid in light direction perception by shading the PAB periodically when the cell rotates helically in lateral light, but stigmaless mutants can also orient with respect to the light direction, and negative phototaxis does not need the presence of the stigma. The PAB is composed of dichroically oriented chromoproteins which is reflected in a pronounced polarotaxis in polarized light. There was a long debate about the potential photoreceptor molecule in Euglena, including carotenoids, flavins and rhodopsins. This discussion was terminated by the unambiguous proof that the photoreceptor is a 400 kDa photoactivated adenylyl cyclase (PAC) which consists of two α- and two β-subunits each. Each subunit possesses two BLUF (Blue Light receptor Using FAD) domains binding FAD, which harvest the light energy, and two adenylyl cyclases, which produce cAMP from ATP. The cAMP has been found to activate one of the five protein kinase s found in Euglena (PK.4). This enzyme in turn is thought to phosphorylate proteins inside the flagellum which result in a change in the flagellar beating pattern and thus a course correction of the cell. The involvements of PAC and protein kinase have been confirmed by RNA interference (RNAi). PAC is responsible for step-up photophobic responses as well as positive and negative phototaxis, but not for the step-down photophobic response, even though the action spectrum of this resembles those for the other two responses. Analysis of several colorless Euglena mutants and the closely related Euglena longa (formerly Astasia longa) confirms the results. Photokinesis shows a completely different action spectrum. Some other Euglena species, such as E. sanguinea and the gliding E. mutabilis, have been investigated, again showing totally different action spectra for phototaxis and photokinesis as well as step-up and step-down photophobic responses.

The unicellular flagellate Euglena gracilis shows positive phototaxis at low-light intensities (<10 W/m(2)) and a negative one at higher irradiances (>10 W/m(2)). Phototaxis is based on blue light-activated adenylyl cyclases, which produce cAMP upon irradiation. In the absence of light the cells swim upward in the water column (negative gravitaxis). The results of sounding rocket campaigns and of a large number of ground experiments led to the following model of signal perception and transduction in gravitaxis of E. gracilis: The body of the cell is heavier than the surrounding medium, sediments and thereby exerts a force onto the lower membrane. Upon deviation from a vertical swimming path mechano-sensitive ion channels are activated. Calcium is gated inwards which leads to an increase in the intracellular calcium concentration and causes a change of the membrane potential. After influx, calcium activates one of several calmodulins found in Euglena, which in turn activates an adenylyl cyclase (different from the one involved in phototaxis) to produce cAMP from ATP. One further element in the sensory transduction chain of both phototaxis and gravitaxis is a specific protein kinase A. We found five different protein kinases A in E. gracilis. The blockage of only one of these (PK.4, accession No. EU935859) by means of RNAi inhibited both phototaxis and gravitaxis, while inhibition of the other four affected neither phototaxis nor gravitaxis. It is assumed that cAMP directly activates this protein kinase A which may in turn phosphorylate a protein involved in the flagellar beating mechanism.

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.

The unicellular freshwater flagellate Euglena gracilis regulates its position in the water column by means of phototactic and gravitactic behavior. Recent experiments have revealed that the cells switch between negative and positive gravitaxis depending upon environmental stimuli such as solar radiation. In this study, the effect of increased salinity on gravitaxis in Euglena gracilis was investigated. In some experiments it was found that salt concentrations up to 5 gL-1 (in some experiments 10 gL-1) increased the motility, velocity and precision of negative gravitactic orientation. Higher salt concentrations decreased all these parameters. At concentrations of about 15 gL-1, cells which did not become immobile, switched from negative to positive gravitaxis. Positive gravitaxis persisted for several hours or even days when the cells were transferred back to standard culture medium. Most of the cells in cultures exposed to salt concentrations above 20 gL-1 lost their motility (partial formation of palmella stages) but recovered when transferred back to standard medium or de-ionised water. Post recovery, the cells showed pronounced positive gravitaxis. Additional investigations on the pigmentation, revealed that the cells showed a complete loss of a carotenoid shoulder in the spectrum, which reappeared when the cells were brought back to standard medium.

The protective effects of Trolox on the inhibition of negative gravitaxis in Euglena gracilis exposed to 200 Gy 60Co gamma-rays were examined using different concentrations (1, 10 and 100 microM). The orientation precision of the negative gravitaxis was quantified using the r-value. A significant decrease in the r-value was observed in gamma-irradiated samples (0.18+/-0.03) compared to those of non-irradiated samples (0.47+/-0.03). There were no significant changes in the r-value of cells exposed to 200 Gy gamma-rays by the addition of 1 or 10 microM of Trolox. A significant increase (0.19) in the r-value of cells exposed to 200 Gy with 100 microM Trolox was observed. The results indicates that Trolox at a concentration of 100 microM protects negative gravitaxis against 60Co gamma-ray irradiation at a dose of 200 Gy. It also suggests that the negative gravitaxis of Euglena gracilis is affected by free radicals.

Human exploration of space and other celestial bodies bears a multitude of challenges. The Earth-bound supply of material and food is restricted, and in situ resource utilisation (ISRU) is a prerequisite. Excellent candidates for delivering several services are unicellular algae, such as the space-approved flagellate Euglena gracilis. This review summarizes the main characteristics of this unicellular organism. Euglena has been exposed on various platforms that alter the impact of gravity to analyse its corresponding gravity-dependent physiological and molecular genetic responses. The sensory transduction chain of gravitaxis in E. gracilis has been identified. The molecular gravi-(mechano-)receptors are mechanosensory calcium channels (TRP channels). The inward gated calcium binds specifically to one of several calmodulins (CaM.2), which, in turn, activates an adenylyl cyclase. This enzyme uses ATP to produce cAMP, which induces protein kinase A, followed by the phosphorylation of a motor protein in the flagellum, initiating a course correction, and, finally, resulting in gravitaxis. During long space missions, a considerable amount of food, oxygen, and water has to be carried, and the exhaled carbon dioxide has to be removed. In this context, E. gracilis is an excellent candidate for biological life support systems, since it produces oxygen by photosynthesis, takes up carbon dioxide, and is even edible. Various species and mutants of Euglena are utilized as a producer of commercial food items, as well as a source of medicines, as it produces a number of vitamins, contains numerous trace elements, and synthesizes dietary proteins, lipids, and the reserve molecule paramylon. Euglena has anti-inflammatory, -oxidant, and -obesity properties. [ABSTRACT FROM AUTHOR]

For long-term manned interplanetary missions it is not feasible to carry the necessary oxygen, food, and water to sustain the astronauts. In addition, the CO2 exhaled by the astronauts has to be removed from the cabin air. One alternative is to utilize photosynthetic organisms to uptake the CO2 and produce oxygen. In addition to higher plants, algae are perfect candidates for this purpose. They also serve to absorb wastes and clean the water. Cyanobacteria can be utilized as food supplement. Early ground-based systems include micro-ecological life support system alternative, closed equilibrated biological aquatic system, and the Biomass Production Chamber. The AQUARACK used the unicellular flagellate Euglena which produced the oxygen for fish in a connected compartment. A number of bioregenerative systems (AQUACELLS, OMEGAHAB) have been built for experiments on satellites. A later experiment was based on a 60-ml closed aquatic ecosystem launched on the Shenzhou 8 spacecraft containing several algae and a small snail living in adjacent chambers. Recently the Eu : CROPIS mission has been launched in a small satellite within a Deutschen Zentrum für Luft- und Raumfahrt (DLR) program. In addition to tomato plants, Euglena is included as oxygen producer. One new approach is to recycle urine on a bacterial filter to produce nitrogen fertilizer to grow vegetables. [ABSTRACT FROM AUTHOR]

Understanding the toxicity of certain potentially toxic compounds on various aquatic organisms allows to assess the impact that these pollutants on the aquatic biota. One source of pollution is the wastewater from hemodialysis. The process of sewage treatment is inefficient in inhibition and removal of pathogenic bacteria resistant to antibiotics in this wastewater. In many countries, such as Brazil, during emergencies, sewage and effluents from hospitals are often dumped directly into waterways without any previous treatment. The objective of this study was to characterize the effluents generated by hemodialysis and to assess the degree of acute and chronic environmental toxicity. The effluents of hemodialysis showed high concentrations of nitrites, phosphates, sulfates, ammonia, and total nitrogen, as well as elevated conductivity, turbidity, salinity, biochemical and chemical oxygen demand, exceeding the thresholds defined in the CONAMA Resolution 430. The samples showed acute toxicity to the green flagellate Euglena gracilis affecting different physiological parameters used as endpoints in an automatic bioassay such as motility, precision of gravitational orientation (r-value), compactness, upward movement, and alignment, with mean EC50 values of recalculate as 76.90 percent (±4.68 percent) of the undiluted effluents. In tests with Daphnia magna, the acute toxicity EC50 was 86.91 percent (±0.39 percent) and a NOEC value of 72.97 percent and a LEOC value 94.66 percent. [Copyright &y& Elsevier]

Water covers more than 70% of the Earth's surface and is the most valuable natural resource on our planet. The rapid increase of human population and industrial development as well as expanding use of chemicals causes pollution to aquatic ecosystems; therefore the demand for methods to determine the damaging and hazardous effects is greater than ever. Ecotoxicology is a field that evaluates the effect of toxins on biological systems. It is a fundamental tool in monitoring pollutants in the environment, toxicity and pollutant identification. The bioassay ECOTOX is an automatic early warning system to monitor probable pollution of freshwater, urban or industrial waste waters or aquatic ecosystems. It uses a real time image analysis of movement and orientation parameters of unicellular flagellate algae, such as the photosynthetic freshwater Euglena gracilis. This organism was found to be severely affected upon exposure to many classes of toxins, such as heavy metals and organic pollutants at low concentrations. In order to expand the application of the instrument to brackish and saline waters, marine flagellates were also evaluated as acknowledged bioassay organisms, e.g. Dunaliella, Prorocentrum and Tetraselmis suecica. The instrument gives the user the opportunity to automatically determine effect-concentration (EC) curves from which several parameters provide important information about the level of toxicity. The NOEC (non observed effect concentration), EC50 and LD (lethal dose) values can be deduced from the data. The system is applicable for short-term (direct exposure to toxins) as well as long-term (incubation time with toxins) risk assessment. The review provides a detailed description of the ECOTOX instruments, about their applications and results obtained so far. [ABSTRACT FROM AUTHOR]