Differential proteomic analyses of green microalga Ettlia sp. at various dehydration levels.

Water deprivation could be a lethal stress for aquatic and aero-terrestrial organisms. Ettlia sp. is a unicellular photosynthetic freshwater microalga. In the present study, proteomic alterations and physiological characteristics of Ettlia sp. were analyzed to comprehend the molecular changes in dehydrated conditions. Varying levels of dehydration were achieved by incubating drained Ettlia sp. in different relative humidity environments for 24  hours. Using a comparative proteomic analysis, 52 differentially expressed protein spots were identified that could be divided into eight functional groups. The PCA analysis of normalized protein expression values demonstrated a clear segregation of protein expression profiles among different dehydration levels. Identified proteins could be grouped into four clusters based on their expression profiles. Proteins relating to photosynthesis comprised the largest group with 25% of the identified proteins that were decreased in dehydrated samples and belonged to cluster I. The photosynthetic activities were measured with rehydrated Ettlia sp. These results revealed that photosynthesis remained inhibited over extended time in response to dehydration. The expressions of reactive oxygen species (ROS) scavenger proteins increased in strong dehydration and were assigned to cluster III. Carbon metabolism proteins were suppressed, which might limit energy consumption, whereas glycolysis was activated at mild dehydration. The accumulation of desiccation-associated late embryogenesis proteins might inhibit the aggregation of housekeeping proteins. DNA protective proteins were expressed higher in the dehydrated state, which might reduce DNA damage, and membrane-stabilizing proteins increased in abundance in desiccation. These findings provide an understanding of Ettlia's adaptation and survival capabilities in a dehydrated state.

Maximizing biomass and lipid production in Ettlia sp. by ultraviolet stress in a continuous culture.

Lipid production in microalgae can be induced by various stress factors. However, stress induced lipid accumulation requires considerable time leading to the decrease in lipid productivity. Here, we attempted to increase the lipid productivity while maintaining the high growth of Ettlia sp. by optimizing nitrogen concentration and UV exposure in a continuous culture. The biomass and lipid productivities of Ettlia sp. cultured with 150 mg N L-1 and UV-A added PAR were 1.67 ±â€¯0.08 g L-1 d-1 and 0.55 ±â€¯0.05 g L-1 d-1, respectively. Lipid productivity and lipid content were around 43.7% and 33.7% higher, respectively in UV-A treatment compared to the control. Moreover, gene-expression patterns related to antioxidant defense and intracellular ROS levels indicated that UV-A affected certain ROS and antioxidants pathways and successfully induced the lipid accumulation in Ettlia sp. This strategy to activate lipid accumulation can be applied in other microalgae without affecting their growth.

Bacterial community enhances flocculation efficiency of Ettlia sp. by altering extracellular polymeric substances profile.

This study examined the effects of a bacterial community and extracellular polymeric substances (EPS) on Ettlia sp. flocculation. The growth rate, flocculation efficiency (FE), bacterial community, and EPS profile of axenic and xenic Ettlia cultures were monitored during 46 days of cultivation. For the xenic culture, with a great abundance of growth-promoting and flocculation-inducing bacteria, the biomass density was 18.75% higher and its FE reached 100% in the mid-stationary phase. Moreover, microscopic observation and a quantitative analysis of the EPS revealed the exclusive presence of long filamentous EPS and more compact structure in the xenic Ettlia culture, possibly explaining its better FE. Notwithstanding, for the axenic culture, despite a lower biomass density and reduced abundance of EPS, its FE reached 92.54% in the mid-stationary phase. Thus, the role of the bacterial community was found to be supportive rather than vital for the high settleability of the self-flocculating Ettlia microalgal culture.

Optimal strategies for bioremediation of nitrate-contaminated groundwater and microalgae biomass production.

Optimizing the mono-cultivation and mixed cultivation of Chlamydomonas reinhardtii, Chlorella vulgaris, and an Ettlia sp. was evaluated for treating nitrate-contaminated groundwater and biomass production. Ettlia sp. showed the highest nutrient assimilation and growth rate among the three microalgae during bioremediation. Light-dark cycle was the effective condition for nutrient removal and COD mitigation by microalgae. Mixed microalgae with a larger presence of the Ettlia sp. exhibited the highest biomass productivity, nitrate-nitrogen, and phosphate-phosphorus removal rates of 0.21 g/L/d, 16.6, and 3.06 mg/L/d, respectively. An N:P mass ratio of 5 was necessary to increase the mixed-microalgal performance. The settling efficiency of the mixed microalgae increased up to 0.55 when using pH modulation during 30 min. Therefore, applying an Ettlia sp.-dominant consortium was the optimum strategy for the bioremediation of nitrate-contaminated groundwater in 3 days.

Bioflocculation in natural and engineered systems: current perspectives.

Microorganisms have the tendency to accumulate at interfaces through the release of extracellular polymeric substances to form aggregates such as films or flocs. This physical association leads to different modes of interactions among cells and the subsequent development of functionally and metabolically diverse consortia. Aggregation of cells in aqueous suspensions often results in the formation of flocs, which are hotspots of enhanced microbial processes. This has important implications for the dynamics of organic and inorganic matter in varied ecosystems. These microbial flocs are not only important components in nutrient turnover, decomposition, and sinking flux but also facilitate contaminant removal and treatment of wastewater and biomass harvesting. Greater insight into the multitude of interactions between microorganisms in flocs would be useful to enhance the efficiency of bioflocculation processes. This review covers the fundamental aspects and outlines the role of bioflocculation in controlled industrial processes and in nature.

Light intensity as major factor to maximize biomass and lipid productivity of Ettlia sp. in CO2-controlled photoautotrophic chemostat.

The optimal culture conditions are critical factors for high microalgal biomass and lipid productivity. To optimize the photoautotrophic culture conditions, combination of the pH (regulated by CO2 supply), dilution rate, and light intensity was systematically investigated for Ettlia sp. YC001 cultivation in a chemostat during 143days. The biomass productivity increased with the increase in dilution rate and light intensity, but decreased with increasing pH. The average lipid content was 19.8% and statistically non-variable among the tested conditions. The highest biomass and lipid productivities were 1.48gL-1d-1 and 291.4mgL-1d-1 with a pH of 6.5, dilution rate of 0.78d-1, and light intensity of 1500µmolphotonsm-2s-1. With a sufficient supply of CO2 and nutrients, the light intensity was the main determinant of the photosynthetic rate. Therefore, the surface-to-volume ratio of a photobioreactor should enable efficient light distribution to enhance microalgal growth.

Phosphorus optimization for simultaneous nitrate-contaminated groundwater treatment and algae biomass production using Ettlia sp.

The effects of phosphorus concentration on the cell growth, nutrient assimilation, photosynthetic parameters, and biomass recovery of Ettlia sp. were evaluated with batch experiments using groundwater, 50mg/L of N-NO3-, and different concentrations of P-PO43-: 0.5, 2.5, 5, and 10mg/L. The maximum biomass productivity and phosphorus removal rate were 0.2g/L/d and 5.95mg/L/d, respectively, with the highest phosphorus concentration of 10mg/L. However, a phosphorus concentration of 5mg/L (N:P=10) was sufficient to ensure an effective nitrogen removal rate of 11mg/L/d, maximum growth rate of 0.88/d, and biomass recovery of 0.72. The appropriate hydraulic retention time was considered as 4days on a large scale to meet the effluent limitation demands of water. While nitrogen depletion had a significant effect on the photosynthetic parameters and ratio of chlorophyll a to dry cell weight during the stationary phase, the effect of phosphorus was negligible during the cultivation.

Evaluation of various techniques for microalgal biomass quantification.

Biomass concentration is one of the most important parameters in the biotechnology processes. Its measurement relies on the physical, chemical or biological properties of the cells. Several techniques were applied in this work to measure the cell concentration of four microalgae: Botryococcussp., Botryococcusbraunii, Chlorella vulgaris, and Ettlia sp. The experiments were performed using samples taken from a chemostat for each strain to provide microalgal cell suspensions in a stable physiological state and concentration. The dry cell weight (DCW) was used as the reference method for the evaluation of other methods. The two commercial sensors used to determine optical density and dielectric permittivity showed a broad effective measurement range up to more than 20gl(-1). A Red-Green-Blue model analysis of microalgal digital images in combination with Fourier equation significantly extended the measurements range up to 6gl(-1). Cell count using a flow cytometer showed a broad range of linearity to DCW in washed samples, but other counting methods using hemocytometer and microscopic automated count were limited. Finally, the oxygen production rate, representing the photosynthetic activity, showed a linear regression with DCW at cell concentrations lower than 1gl(-1).