Growth on ATP Elicits a P-Stress Response in the Picoeukaryote Micromonas pusilla.

The surface waters of oligotrophic oceans have chronically low phosphate (Pi) concentrations, which renders dissolved organic phosphorus (DOP) an important nutrient source. In the subtropical North Atlantic, cyanobacteria are often numerically dominant, but picoeukaryotes can dominate autotrophic biomass and productivity making them important contributors to the ocean carbon cycle. Despite their importance, little is known regarding the metabolic response of picoeukaryotes to changes in phosphorus (P) source and availability. To understand the molecular mechanisms that regulate P utilization in oligotrophic environments, we evaluated transcriptomes of the picoeukaryote Micromonas pusilla grown under Pi-replete and -deficient conditions, with an additional investigation of growth on DOP in replete conditions. Genes that function in sulfolipid substitution and Pi uptake increased in expression with Pi-deficiency, suggesting cells were reallocating cellular P and increasing P acquisition capabilities. Pi-deficient M. pusilla cells also increased alkaline phosphatase activity and reduced their cellular P content. Cells grown with DOP were able to maintain relatively high growth rates, however the transcriptomic response was more similar to the Pi-deficient response than that seen in cells grown under Pi-replete conditions. The results demonstrate that not all P sources are the same for growth; while M. pusilla, a model picoeukaryote, may grow well on DOP, the metabolic demand is greater than growth on Pi. These findings provide insight into the cellular strategies which may be used to support growth in a stratified future ocean predicted to favor picoeukaryotes.

Boosting TAG Accumulation with Improved Biodiesel Production from Novel Oleaginous Microalgae Scenedesmus sp. IITRIND2 Utilizing Waste Sugarcane Bagasse Aqueous Extract (SBAE).

This investigation utilized sugarcane bagasse aqueous extract (SBAE), a nontoxic, cost-effective medium to boost triacylglycerol (TAG) accumulation in novel fresh water microalgal isolate Scenedesmus sp. IITRIND2. Maximum lipid productivity of 112 ± 5.2 mg/L/day was recorded in microalgae grown in SBAE compared to modified BBM (26 ± 3 %). Carotenoid to chlorophyll ratio was 12.5 ± 2 % higher than in photoautotrophic control, indicating an increase in photosystem II activity, thereby increasing growth rate. Fatty acid methyl ester (FAME) profile revealed presence of C14:0 (2.29 %), C16:0 (15.99 %), C16:2 (4.05 %), C18:0 (3.41 %), C18:1 (41.55 %), C18:2 (12.41), and C20:0 (1.21 %) as the major fatty acids. Cetane number (64.03), cold filter plugging property (-1.05 °C), and oxidative stability (12.03 h) indicated quality biodiesel abiding by ASTM D6751 and EN 14214 fuel standards. Results consolidate the candidature of novel freshwater microalgal isolate Scenedesmus sp. IITRIND2 cultivated in SBAE, aqueous extract made from copious, agricultural waste sugarcane bagasse to increase the lipid productivity, and could further be utilized for cost-effective biodiesel production.

Community structure and nutrient level control the tolerance of autotrophic biofilm to silver contamination.

Autotrophic biofilms are complex and fundamental biological compartments of many aquatic ecosystems. Since microbial species differ in their sensitivity to stressors, biofilms have long been proposed for assessing the quality of aquatic ecosystems. Among the many stressors impacting aquatic ecosystems, eutrophication and metal pollution are certainly the most common. Despite that these stressors often occur together, their effects on biofilms have been far much studied separately than interactively. In this study, we evaluated the interactive effects of silver (Ag), a reemerging contaminant, and phosphorus (P), a nutrient often associated with freshwater eutrophication, on the structure and functioning of two types of autotrophic biofilms, one dominated by diatoms and another one dominated by cyanobacteria. We hypothesized that P would alleviate the toxic effects of Ag, either directly, through the contribution of P in metal detoxification processes, or indirectly, through P-mediated shifts in biofilm community compositions and associated divergences in metal tolerance. Results showed that Ag impacted biofilm community structure and functioning but only at unrealistic concentrations (50 µg/L). P availability led to significant shifts in biofilm community composition, these changes being more pronounced in diatom- than those in cyanobacteria-dominated biofilm. In addition, P tended to reduce the impact of Ag but only for the cyanobacteria-dominated biofilm. More generally, our results highlight the preponderant role of the initial community structure and nutrient level on biofilm response to metallic pollutants.

Isolation and characterization of microalgae for biodiesel production from seawater.

As green marine microalgae isolated from local seawater in Tianjin, China, Nannochloropsis gaditana Q6 was tolerant to the variation of salinity with the highest biomass and lipid concentration in natural seawater medium. Although this strain could grow mixotrophically with glycerol, the narrow gap between mixotrophic and autotrophic cultivation suggested that autotrophic cultivation was the optimal trophic type for N. gaditana Q6 growth. In addition, strain Q6 was more sensitive to the variance of NH4HCO3 concentration than NaH2PO4 concentration. Consequently, the lipid production could be maximized by the two-stage cultivation strategy, with an initial high NH4HCO3 concentration for biomass production followed by low NH4HCO3 concentration for lipid accumulation.

Screening and characterization of oleaginous Chlorella strains and exploration of photoautotrophic Chlorella protothecoides for oil production.

The growth and oil production of nine Chlorella strains were comparatively assessed and Chlorellaprotothecoides CS-41 demonstrated the greatest lipid production potential. The effects of different nitrogen forms and concentrations, phosphorus concentrations and light intensities on growth and oil production were studied in laboratory columns. C. protothecoides CS-41 accumulated lipids up to 55% of dry weight, with triacylglycerol and oleic acid being 71% of total lipids and 59% of total fatty acids, respectively. High biomass and lipid productivities were achieved in outdoor panel PBRs, up to 1.25 and 0.59 g L(-1) day(-1), or 44. 1 and 16.1 g m(-2) day(-1), respectively. A two-stage cultivation strategy was proposed to enhance the algal biomass and lipid production. This is the first comprehensive investigation of both indoor and outdoor photoautotrophic C. protothecoides cultures for oil production, and C. protothecoides CS-41 represents a promising biofuel feedstock worthy of further exploration.

A new biological phosphorus removal process in association with sulfur cycle.

Hong Kong has practiced seawater toilet flushing since 1958, saving 750,000 m(3) freshwater every day. A high sulfate-to-COD ratio (>1.25 mg SO4/mg COD) in the saline sewage resulting from this practice has enabled us to develop the Sulfate reduction Autotrophic denitrification and Nitrification Integrated (SANI(®)) process with minimal sludge production. This study seeks to expand the SANI process into an enhanced biological phosphorus removal (EBPR) process. A sulfur cycle associated EBPR was explored in an alternating anaerobic/oxygen-limited aerobic sequencing batch reactor with acetate fed as sole electron donor and sulfate as sulfur source at a total organic carbon to sulfur ratio of 1.1-3.1 (mg C/mg S). Phosphate uptake and polyphosphate formation was observed in this reactor that sustained high phosphate removal (20 mg P/L removed with 320 mg COD/L). This new EBPR process was supported by six observations: 1) anaerobic phosphate release associated with acetate uptake, poly-phosphate hydrolysis, poly-hydroxyalkanoate (PHA) (and poly-S(2-)/S(0)) formation and an "aerobic" phosphate uptake associated with PHA (and poly-S(2-)/S(0)) degradation, and polyphosphate formation; 2) a high P/VSS ratio (>0.16 mg P/mg VSS) and an associated low VSS/TSS ratio (0.75) characteristic of conventional PAOs; 3) a lack of P-release and P-uptake with formaldehyde inactivation and autoclaved sterilized biomass; 4) an absence of chemical precipitated P crystals as determined by XRD analysis; 5) a sludge P of more than 90% polyphosphate as determined by sequential P extraction; and 6) microscopically, observed PHA, poly-P and S globules in the biomass.

Periphyton responses to nutrient and atrazine mixtures introduced through agricultural runoff.

Agricultural runoff often contains pollutants with antagonistic impacts. The individual influence of nutrients and atrazine on periphyton has been extensively studied, but their impact when introduced together and with multiple agricultural pollutants is less clear. We simulated a field-scale runoff pulse into a riverine wetland that mimicked pollutant composition typical of field runoff of the Mississippi River Alluvial Plain. Periphyton biomass and functional responses were measured for 2 weeks along a 500 m section. Additionally, laboratory chamber assays were used to identify potential periphyton changes due to nutrients, atrazine, and their interactions. Generally, nutrients stimulated, and atrazine reduced chlorophyll a (Chl a) in chambers. In the wetland, nutrient and atrazine relationships with periphyton were weaker, and when found, were often opposite of trends in chambers. Total nitrogen (TN) was inversely related to Chl a, and total phosphorus was inversely related to respiration (R) rates. Atrazine (10-20 µg L(-1) in the wetland) had a positive relationship with ash-free dry mass (AFDM), and weakened the relationship between TN and AFDM. Wetland periphyton biomass was better correlated to total suspended solids than nutrients or atrazine. Periphyton function was resilient as periphyton gross primary production (GPP)/R ratios were not strongly impacted by runoff. However, whole-system GPP and R decreased over the 2-week period, suggesting that although periphyton metabolism recovered quickly, whole-system metabolism took longer to recover. The individual and combined impacts of nutrients and atrazine in complex pollutant mixtures can vary substantially from their influence when introduced separately, and non-linear impacts can occur with distance downstream of the pollutant introduction point.

Removal of highly elevated nitrate from drinking water by pH-heterogenized heterotrophic denitrification facilitated with ferrous sulfide-based autotrophic denitrification.

The performance of acetic acid-supported pH-heterogenized heterotrophic denitrification (HD) facilitated with ferrous sulfide-based autotrophic denitrification (AD) was investigated in upflow activated carbon-packed column reactors for reliable removal of highly elevated nitrate (42 mg NO(3)-Nl(-1)) in drinking water. The use of acetic acid as substrate provided sufficient internal carbon dioxide to completely eliminate the need of external pH adjustment for HD, but simultaneously created vertically heterogenized pH varying from 4.8 to 7.8 in the HD reactor. After 5-week acclimation, the HD reactor developed a moderate nitrate removal capacity with about one third of nitrate removal occurring in the acidic zone (pH 4.8-6.2). To increase the treatment reliability, acetic acid-supported HD was operated under 10% carbon limitation to remove >85% of nitrate, and ferrous sulfide-based AD was supplementally operated to remove residual nitrate and formed nitrite without excess of soluble organic carbon, nitrite or sulfate in the final effluent.