pH effects on growth and lipid accumulation of the biofuel microalgae Nannochloropsis salina and invading organisms

Biofuels derived from non-crop sources, such as microalgae, offer their own advantages and limitations. Despite high growth rates and lipid accumulation, microalgae cultivation still requires more energy than it produces. Furthermore, invading organisms can lower efficiency of algae production. Simple environmental changes might be able to increase algae productivity while minimizing undesired organisms like competitive algae or predatory algae grazers. Microalgae are susceptible to pH changes. In many production systems, pH is kept below 8 by CO₂ addition. Here, we uncouple the effects of pH and CO₂ input, by using chemical pH buffers and investigate how pH influences Nannochloropsis salina growth and lipid accumulation as well as invading organisms. We used a wide range of pH levels (5, 6, 7, 8, 9, and 10). N. salina showed highest growth rates at pH 8 and 9 (0.19?±?0.008 and 0.19?±?0.011, respectively; mean ± SD). Maximum cell densities in these treatments were reached around 21 days into the experiment (95.6?×?10⁶?±?9?×?10⁶ cells mL^?1 for pH 8 and 92.8?×?10⁶?±?24?×?10⁶ cells mL^?1 for pH 9). Lipid accumulation of unbuffered controls were 21.8?±?5.8 % fatty acid methyl esters content by mass, and we were unable to trigger additional significant lipid accumulation by manipulating pH levels at the beginning of stationary phase. Ciliates (grazing predators) occurred in significant higher densities at pH 6 (56.9?±?39.6?×?10⁴ organisms mL^?1) than higher pH treatments (0.1–6.8?×?10⁴ organisms mL^?1). Furthermore, the addition of buffers themselves seemed to negatively impact diatoms (algal competitors). They were more abundant in an unbuffered control (12.7?±?5.1?×?10⁴ organisms mL^?1) than any of the pH treatments (3.6–4.7?×?10⁴ organisms mL^?1). In general, pH values of 8 to 9 might be most conducive to increasing algae production and minimizing invading organisms. CO₂ addition seems more valuable to algae as an inorganic carbon source and not as an essential mechanism to reduce pH.     

Abundance and primary production of filamentous green algae Zygogonium ericetorum in an extremely acid (pH 2.9) mining lake and its impact on alkalinity generation

1. In extremely acid mining lakes, benthic filamentous green algae (Zygnemataceae, Chlorophyta) thrive as effective competitors for limited carbon (C). These algae could supply C for microbial‐mediated benthic alkalinity generation. However, biomass, productivity and impact of the acidobiontic filamentous green algae at pH ≤3 have not previously been determined. 2. Periphytic filamentous green algae was mapped by harvesting their biomass from 85 1 × 1 m quadrats in mining lake Grünewalder Lauch. Zygogonium ericetorum colonised water depths between 1.6 and 10.5 m covering 88% of total area. Biomass peaked at 5–6 m depth. Total Zygogonium biomass amounted to 72.2 t dry weight for the whole lake (0.94 km²), which corresponds to 16.1 t C and the accumulation of primary production from 2.2 years. 3. Growth of Zygogonium is moderately N, C and extremely P deficient, and seriously stressed by high rates of Fe deposition during summer. Consequently, net primary production (NPP) of Zygogonium, calculated from measured photosynthesis versus irradiance characteristics and calculated underwater irradiance (0.13 g C m^?2 year^?1) and in situ oxygen measurements (7.8 g C m^?2 year^?1), corresponds to only 0.3% and 18.1% of pelagic NPP. 4. Neither pelagic nor benthic Zygogonium primary production can supply enough C for efficient acidity removal. However, at rates of benthic NPP in summer of 21.4 mg C m^?2 day^?1, Zygogonium contributed 26% of the C equivalents to remove acidity associated with ferric iron, contributing at least seasonally to efficient alkalinity generation.     

Chemical composition and ethanol production potential of Thai seaweed species

This study evaluated the potential use of several Thai seaweed species for ethanol production. The high biomass of the green algae Ulva intestinalis and Rhizoclonium riparium and the red algae Gracilaria salicornia and Gracilaria tenuistipitata in an earthen pond culture led us to select these species for our study. The seaweed species were analyzed for chemical composition, resulting in ash contents of 37.62?±?0.15 % and fiber of 11.93?±?0.16 %, with the highest values in R. riparium. Low lipid values were found in all species, with the highest value (p?<?0.05) in G. salicornia (1.69?±?0.07 %) and the lowest in R. riparium (0.28?±?0.01 %) and G. tenuistipitata (0.26?±?0.01 %). The highest carbohydrate contents were found in G. tenuistipitata (54.89 %), and the lowest were in R. riparium (29.53 %). G. tenuistipitata (8.58?±?0.36 %) and U. intestinalis (8.24?±?0.28 %) had higher sulfate contents compared with G. salicornia (4.69?±?0.04 %) and R. riparium (1.97?±?0.20 %). The monosugar algal tissue components were analyzed by HPLC; rhamnose, xylose, fucose, arabinose, mannose, glucose, and galactose were used as reference sugars. Total sugar was found to be highest in G. tenuistipitata (98.21 %). Arabinose, glucose, and galactose were the main sugar components in all species. Glucose obtained from G. tenuistipitata (6.55 %) and R. riparium (6.52 %) was higher than in G. salicornia (0.27 %) and U. intestinalis (2.78 %). G. tenuistipitata fermentation gave a higher yield of ethanol (4.17?×?10^?3 g ethanol g^?1 sugars; 139.12 μg ethanol g^?1 glucose) than R. riparium (0.086?×?10^?3 g ethanol g^?1 sugars; 33.84 μg ethanol g^?1 glucose), U. intestinalis (0.074?×?10^?3 g ethanol g^?1 sugars; 9.98 μg ethanol g^?1 glucose), and G. salicornia (0.031?×?10^?3 g ethanol g^?1 sugars; 1.43 μg ethanol g^?1 glucose).     

Elicitor mediated enhancement of wedelolactone in cell suspension culture of <Emphasis Type="Italic">Eclipta alba</Emphasis> (L.) Hassk

The present research focused on enhancing the production of wedelolactone through cell suspension culture (CSC) in Eclipta alba (L.) Hassk. With an aim of attaining a sustainable CSC, various plant growth regulators, elicitors and agitation speed were examined. Nodal segments of in vitro propagated plantlets induced the maximum percentage (93.47?±?0.61%) of callus inoculated on Murashige and Skoog (MS) medium fortified with picloram (2 mg L^?1). The growth kinetics of CSC exhibited a sigmoid pattern with a lag phase (0–6 days), a log phase (6–18 days), a stationary phase (18–24 days) and then death phase thereafter. The highest biomass accumulation in CSC with 7.09?±?0.06 g 50 mL^?1 fresh weight, 1.52?±?0.02 g 50 mL^?1 dry cell weight, 1.34?±?0.01?×?10⁶ cell mL^?1 total cell count and 57.00?±?0.58% packed cell volume was obtained in the liquid MS medium supplemented with 1.5 mg L^?1 picloram plus 0.5 mg L^?1 kinetin at 120 rpm. High performance thin layer chromatography confirmed that yeast extract (biotic elicitor) at 150 mg L^?1 accumulated more CSC biomass with 1.22-fold increase in wedelolactone (288.97?±?1.94 µg g^?1 dry weight) content in comparison to the non-elicited CSC (237.78?±?0.04 µg g^?1 dry weight) after 120 h of incubation. Contrastingly, methyl jasmonate (abiotic elicitor) did not alter the biomass but increased the wedelolactone content (259.32?±?1.06 µg g^?1 dry weight) to an extent of 1.09-fold at 100 µM. Complete plantlet regeneration from CSC was possible on MS medium containing N⁶-benzyladenine (0.75 mg L^?1) and abscisic acid (0.5 mg L^?1). Thus, the establishment of protocol for CSC constitutes the bases for future biotechnological improvement studies in this crop.     

Growing microalgae as aquaculture feeds on twin-layers: a novel solid-state photobioreactor

Four strains of marine microalgae commonly used as live feeds in hatcheries (Isochrysis sp. T.ISO, Tetraselmis suecica, Phaeodactylum tricornutum, Nannochloropsis sp.) were grown in a novel solid-state photobioreactor, the twin-layer system. Microalgae were immobilized by self adhesion to vertically oriented twin-layer modules which consisted of two different types of ultrathin layers, a macroporous source layer (glass fiber nonwoven) through which the culture medium was transported by gravity flow, and a microporous substrate layer (plain printing paper) which carried the algae on both surfaces of the source layer. This simple open cultivation system effectively separated the immobilized microalgae from the bulk of the growth medium and permitted prolonged cultivation of microalgae with average biomass yields of 10–15 g dry weight m^?2 growth area after 14–25 days of cultivation. Algal biomass was harvested as fresh weight (with 72–84 % water content) without the need to pre-concentrate algae. No aeration or external CO₂ supply was necessary, and due to the microporous substrate layer, no eukaryotic contaminations were observed during the experiment. All experiments were conducted in Germany under greenhouse conditions with natural sunlight. Small-scale growth experiments performed under the same conditions revealed that growth over most of the experimental period (24 days) was linear in all tested algae with growth rates (dry weight per square meter growth area) determined to be 0.6 g ?m^?2?day^?1 (Isochrysis), 0.8 g? m^?2?day^?1 (Nannochloropsis), 1.5 g ?m^?2?day^?1 (Tetraselmis), and 1.8 g? m^?2?day^?1 (Phaeodactylum). Due to its cost-effective construction and with further optimisation of design and productivity at technical scales, the twin-layer system may provide an attractive alternative to methods traditionally used to cultivate live microalgae.     

Evaluation of the in vitro methods for micropropagation of Chondracanthus acicularis (Roth) Fredericq (Gigartinales, Rhodophyta): tissue culture and production of protoplasts

Tissue was cultured and protoplasts isolated from the carrageenophyte Chondracanthus acicularis with the aim of developing micropropagation as an alternative to harvesting raw material from natural beds. Both adventitious shoots and filamentous calluses were induced by tissue culture on medium solidified with 0.4–1 % (w/v) agar. Adventitious shoots were mainly produced from discoid bases while filamentous calluses were mainly induced from basal zones and sub-apical explants. A gradient of the regeneration ability was observed from the top to the bottom of the thallus. The discoid base was the most reactive explant and produced the highest number of adventitious shoots compared to basal zones and sub-apical explants, irrespective of the concentration of agar. Protoplasts were isolated enzymatically from the whole thallus using a combination of cellulase R-10 Onozuka, macerozyme R-10, and crude extract of the gland gut of algivorous molluscs. The highest mean yield of protoplasts (1.2?×?10⁶ protoplasts g^?1 fresh weight) was obtained after 16 h of digestion with an enzyme mixture containing 2 % (w/v) cellulase R-10, 1 % (w/v) macerozyme R-10 Onozuka, 4 % (v/v) crude extract of gut gland of Haliotis, 0.8 M mannitol, 50 mM sodium citrate, 0.3 % (w/v) bovine serum albumin. Depending on the conditions, mean protoplast yields ranged from 3.14?×?10⁵ to 1.2?×?10⁶ protoplasts g^?1 fresh weight. Different factors (storage duration, mannitol, sodium citrate, crude extract of the gland gut of algivorous molluscs) were tested to improve the yield of protoplasts but none has a significantly effect.     

Spatiotemporal Variation of Bacterial Assemblages in a Shallow Subtropical Coastal Lagoon in Southern Brazil

A study on the bacterioplankton of Conceição Lagoon (27°34′ S–48°27′ W), Southern Brazil, was carried out in July 2005 (austral winter) and January 2006 (austral summer) to characterize the bacterial spatiotemporal distribution and to determine the heterotrophic and photoautotrophic bacterial dominance in hypoxic/oxic stratified waters. Bacterial abundance increased significantly (p?5 (winter) to 3.21?×?10⁶ cells mL^?1 (summer), heterotrophic coccus/rod-shaped (HCR) cells from 7.00?×?10⁴ to 3.60?×?10⁶ cells mL^?1, and heterotrophic filamentous (HF) bacteria from 2.90?×?10³ to 2.74?×?10⁵ cells mL^?1. Bacterial biovolumes also increased in summer with mean biovolumes of CCY ranging from 0.38 to 1.37 μm³, HCR cells from 0.31 to 1.12 μm³, and HF from 3.32 to 11.34 μm³. Principal component analysis showed that salinity, temperature, and light were the abiotic factors that better explained the temporal variability of bacterial assemblages. Bacterial heterotrophy dominated in the lagoon, excepted by the southern and part of central sector in January 2006, when autotrophic-dominated microbial community occurred. Spatially, bacterial assemblages were influenced by nutrient gradient, oxygen, and salinity with a positive relationship between biovolumes and nutrients and a negative relationship between abundance of coccus cyanobacteria and nutrients. area revealed a singular temporal pattern with hypoxic bottom waters in winter and oxygen-rich waters appearing in summer related with the availability of light and predominant microbes. Thus, oxygen consumption/production is likely to be regulated by the amount of light reaching the bottom, stimulating the production of oxygen by oxygenic phototrophs.     

Cultivation of the two perennial brown algae Ecklonia cava and E. stolonifera for abalone feeds in Korea

Ecklonia cava and Ecklonia stolonifera are perennial brown algae that form sea forests off the coast of Korea. Both species are cultured to supply a summer feed for the abalone industry. Recent expansion of the abalone industry in Korea has been bringing an increase in demand for fresh algal supply. Zoospores of the two algae were seeded in October 2006 on seed frames coiled with 100 m of seed fibers. After 2 months of indoor culture and 2 months of intermediate culture, growth and production of the two algae were compared during their main cultivation period from March 2007 to June 2008, in the culture ground in Wando, Korea (34°26′18.68″ N, 127°05′43.88″ E), in situ. Growth rate of E. cava and E. stolonifera was 1.058 and 3.089 mm day^?1, respectively. The mean production of E. stolonifera obtained from the culture ropes was ca. 12 kg wet wt. m^?1 of culture rope while production of E. cava was ca. 3 kg wet wt. m^?1 of culture rope. The difference in production was attributed from the different growth strategies of the two algae, with only E. stolonifera being able to regenerate blades from the holdfast. The ability to regenerate blades from the holdfast therefore makes E. stolonifera the preferred species for biomass production for abalone feeds. In a 120-day feeding experiment, growth rate, weight gain, and survival rate of abalone showed that E. cava and E. stolonifera feeds could provide an alternative feed to Saccharina japonica during summer months.     

Combined effect of five single nucleotide polymorphisms related to IL23/Th17 pathway in the risk of psoriasis

Psoriasis is a common immune-mediated inflammatory skin disease with strong genetic components, in which the IL23/Th17 pathway has been implicated. To explore the effective role in psoriasis, we genotyped five single nucleotide polymorphisms in genes related to IL23/Th17 pathway in 14,929 Han Chinese samples. A Bonferroni-corrected significant single-variant association was identified between rs1512970 within IL21 (odds ratio (OR)?=?1.07, 95 % confidence interval (CI)?=?1.02–1.13, P?=?4.94?×?10^?03). We failed to validate rs744166 (OR?=?1.06, 95 % CI?=?1.01–1.11, P?=?1.52?×?10^?02) and other three SNPs (P?=?2.48?×?10^?01?～?1.27?×?10^?02) to meet the single-variant association significance threshold. However, we found that their combined effect substantially contributed to the risk of psoriasis in our sample (P?=?3.91?×?10^?07) and the highest score group conferred the largest risk effect size (OR?=?1.22, 95 % CI?=?1.11–1.34, P?=?1.85?×?10^?05). Our results implicate the ethnic heterogeneity in the susceptibility of psoriasis and suggest common variants with weak effect in IL23/Th17 pathway, which do not show significance in conventional single-variant association study, may contribute to the risk of psoriasis. This study sheds light on the important role of IL23/Th17 pathway in the susceptibility of psoriasis.     

Factors correlating with deterioration of giant kelp Macrocystis pyrifera (Laminariales, Heterokontophyta) in an aquarium setting

Survival of the giant kelp, Macrocystis pyrifera (Linnaeus) C. Agardh, in its natural habitat is governed by abiotic and biotic factors such as temperature, light, nutrients, current velocity, and predators. Factors affecting the survival of the alga in an aquarium setting, however, have not been investigated. The National Museum of Marine Biology and Aquarium (NMMBA), in subtropical Taiwan, is the only aquarium in the world that displays giant kelp that does not have naturally occurring specimens in nearby waters. Giant kelp displayed in aquaria often deteriorates within a 3-month period, yet the cause of this mortality is unknown. We investigated abiotic and biotic parameters affecting survival of giant kelp in aquaria over a 3-month period. The results indicated that temperature, salinity, pH, light, and nutrient concentrations did not affect giant kelp survival. However, the massive proliferation of epiphytic diatoms on kelp blades (from 7?×?10² cells cm^?2 initially to 3?×?10⁴ cells cm^?2 after 1 month) was identified as being the most likely candidate affecting survival of giant kelp in an aquarium setting. Potential factors that may stimulate epiphyte proliferation include lack of epiphytic algae control via predation, high nutrient concentrations, a weak current, and a generally stable environment.     

Fate of radiocesium in freshwater aquatic plants and algae in the vicinity of the Fukushima Daiichi nuclear power plant

The behavior of radiocesium (¹³⁷Cs) in aquatic plants (five species) and algae (three genera) grown in either a river (one sampling point) or pond (four sampling points) in the vicinity of the Fukushima Daiichi nuclear power plant was investigated. The ¹³⁷Cs concentration of <0.45-μm fractions of water taken from the river and ponds was between 5.01 × 10^?1 and 2.98 Bq/L, while that of sediment was between 4.85 × 10³ and 5.72 × 10⁴ Bq/kg dry weight. The ratio of ¹³⁷Cs concentration of sediment/water in ponds was ~10⁴. The sediment-to-plant transfer factor (TF) [(¹³⁷Cs concentration Bq/kg dry weight_plant) × (¹³⁷Cs concentration Bq/kg dry weight_sediment)^?1] was also measured. For aquatic plants, the highest value was 5.55 for Potamogeton crispus from the river, while the lowest was 3.34 × 10^?2 for P. distinctus from a pond. There were significant differences in values between aquatic plants belonging to the same genus. The water-to-plant TF [(¹³⁷Cs concentration Bq/kg dry weight_plant) × (¹³⁷Cs concentration Bq/L_water)^?1] of filamentous algae (Spirogyra sp.) and cyanobacteria (coexisting Anabaena sp. and Microcystis sp.) were 2.39 × 10³ and 1.26 × 10³, respectively. The ¹³⁷Cs concentration of cyanobacteria in pond water was 4.87 × 10^?1 Bq/L, which was the same order of magnitude as the ¹³⁷Cs concentration of pond water. Enrichment of ¹³⁷Cs in cyanobacteria was not observed.     

<Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> Spore Production Under Solid-State Fermentation of Lignocellulosic Residues

This study was conducted to elucidate cultivation conditions determining Bacillus amyloliquefaciens B-1895 growth and enhanced spore formation during the solid-state fermentation (SSF) of agro-industrial lignocellulosic biomasses. Among the tested growth substrates, corncobs provided the highest yield of spores (47?×?10¹⁰ spores g^?1 biomass) while the mushroom spent substrate and sunflower oil mill appeared to be poor growth substrates for spore formation. Maximum spore yield (82?×?10¹⁰ spores g^?1 biomass) was achieved when 15 g corncobs were moistened with 60 ml of the optimized nutrient medium containing 10 g peptone, 2 g KH₂PO₄, 1 g MgSO₄·7H₂O, and 1 g NaCl per 1 l of distilled water. The cheese whey usage for wetting of lignocellulosic substrate instead water promoted spore formation and increased the spore number to 105?×?10¹⁰ spores g^?1. Addition to the cheese whey of optimized medium components favored sporulation process. The feasibility of developed medium and strategy was shown in scaled up SSF of corncobs in polypropylene bags since yield of 10?×?10¹¹ spores per gram of dry biomass was achieved. In the SSF of lignocellulose, B. amyloliquefaciens B-1895 secreted comparatively high cellulase and xylanase activities to ensure good growth of the bacterial culture.     

Exploiting Phosphate-Starved cells of <Emphasis Type="Italic">Scenedesmus</Emphasis> sp. for the Treatment of Raw Sewage

Phosphate depletion is one of the favorable ways to enhance the sewage water treatment with the algae, however, detailed information is essential with respect to internal phosphate concentration and physiology of the algae. The growth rate of the phosphate-starved Scenedesmus cells was reduced drastically after 48 h. Indicating cells entered in the stationary phase of the growth cycle. Fourier Transform Infrared analysis of phosphate-starved Scenedesmus cells showed the reduction in internal phosphate concentration and an increase in carbohydrate/phosphate and carbohydrate/lipid ratio. The phosphate-starved Scenedesmus cells, with an initial cell density of, 1 × 10⁶ cells mL^?1 shows 87% phosphate and 100 % nitrogen removal in 24 h. The normal Scenedesmus cells need approximately 48 h to trim down the nutrients from wastewater up to this extent. Other microalgae, Ankistrodesmus, growth pattern was not affected due to phosphate starvation. The cells of Ankistrodesmus was able to reduce 71% phosphate and 73% nitrogen within 24 h, with an initial cell density of, 1 × 10⁶ cells mL^?1.     

Sea ice microbial dynamics over an annual ice cycle in Prydz Bay, Antarctica

Microbial community dynamics within the fast sea ice of Prydz Bay (68°S?78°E) were investigated over an annual cycle at two sites (1 and 3?km offshore) between April and November 2008. There are few long-term sea ice studies, and few that cover the phase of winter darkness when autotrophic processes are curtailed. Mean chlorophyll a concentrations in the ice column ranged between 0.76 and 44.8?μg?L^?1 at the 1-km site (Site 1) and 3.11–144.6?μg?L^?1 at the 3-km site (Site 2). Highest chlorophyll a usually occurred at the base of the ice. Bacterial concentrations ranged between 0.30 and 2.08?×?10⁸?cells?L^?1, heterotrophic nanoflagellates (HNAN) between 0.21?×?10⁵ and 2.98?×?10⁵?cells?L^?1 and phototrophic nanoflagellates (PNAN) 0–1.06?×?10⁵?cells?L^?1. While HNAN occurred throughout the year, PNAN were largely absent in winter. Dinoflagellates were a conspicuous and occasionally an abundant element of the community (maximum 17,460?cells?L^?1), while ciliates were sparse. The bacterial community showed considerable morphological diversity with a dominance of filamentous forms. Bacterial production continued throughout the year ranging between 0 and 22.92?μg?C?L^?1?day^?1 throughout the ice column. Lowest rates occurred between late June and early August. The sea ice sustained an active and diverse microbial community through its annual extent. The data suggest that during winter darkness the microbial community is dominated by heterotrophic processes, sustained by a pool of dissolved organic carbon.     

Strong interactions between stoichiometric constraints and algal defenses: evidence from population dynamics of Daphnia and algae in phosphorus-limited microcosms

The dynamic interactions among nutrients, algae and grazers were tested in a 2 × 3 factorial microcosm experiment that manipulated grazers (Daphnia present or absent) and algal composition (single species cultures and mixtures of an undefended and a digestion-resistant green alga). The experiment was run for 25 days in 10-L carboys under mesotrophic conditions that quickly led to strong phosphorus limitation of algal growth (TP ? 0.5 μM, N:P 40:1). Four-day Daphnia juvenile growth assays tested for Daphnia P-limitation and nutrient-dependent or grazer-induced algal defenses. The maximal algal growth rate of undefended Ankistrodesmus (mean ± SE for three replicate microcosms; 0.92 ± 0.02 day^?1) was higher than for defended Oocystis (0.62 ± 0.03 day^?1), but by day 6, algal growth was strongly P-limited in all six treatments (molar C:P ratio >900). The P-deficient algae were poor quality resources in all three algal treatments. However, Daphnia population growth, reproduction, and survival were much lower in the digestion-resistant treatment even though growth assays provided evidence for Daphnia P-limitation in only the undefended and mixed treatments. Growth assays provided little or no support for simple threshold element ratio (TER) models that fail to consider algae defenses that result in viable gut passage. Our results show that strong P-limitation of algal growth enhances the defenses of a digestion-resistant alga, favoring high abundance of well-defended algae and energy limitation of zooplankton growth.     

Production of microsclerotia by Brazilian strains of Metarhizium spp. using submerged liquid culture fermentation

We investigated the potential production and desiccation tolerance of microsclerotia (MS) by Brazilian strains of Metarhizium anisopliae (Ma), M. acridum (Mc) and M. robertsii (Mr). These fungi were grown in a liquid medium containing 16 g carbon l^?1 with a carbon:nitrogen ratio of 50:1. One hundred milliliters cultures were grown in 250 ml Erlenmeyer flasks in a rotary incubator shaker at 28 °C and 200 rpm for 5 days. Five-day-old MS were harvested, mixed with diatomaceous earth (DE) and air-dried for 2 days at 30 °C. The air-dried MS–DE granular preparations were milled by mortar + pestle and stored in centrifuged tubes at either 26 or ?20 °C. Desiccation tolerance and conidia production were assessed for dried MS granules by measuring hyphal germination after incubation for 2 days on water agar plates at 26 °C and for conidia production following 7 days incubation. Yields of MS by all strains of Metarhizium were 6.1–7.3 × 10⁶ l^?1 after 3 days growth with maximum MS yields (0.7–1.1 × 10⁷ l^?1) after 5 days growth. No differences in biomass accumulation were observed after 3 days growth, whereas Ma-CG168 showed the highest biomass accumulation after 5 days growth. Dried MS–DE preparations of all fungal strains were equally tolerant to desiccation (≥93 % germination) and the highest conidia production was obtained by MS granules of Mc-CG423 (4 × 10⁹ conidia g^?1). All MS granules showed similar stability after storage at either 26 or ?20 °C for 3.5 months.     

Impact of seasonal changes in stream metabolism on nitrate concentrations in an urban stream

Nitrate (NO₃ ^?) dynamics in urban streams differ from many natural streams due to stormwater runoff, sewage inputs, decreased groundwater discharge, often limited hyporheic exchange, increased primary productivity, and limited carbon input. We investigated NO₃ ^? dynamics in a first-order urban stream in Syracuse, NY, which has urbanized headwaters and a geomorphologically natural downstream section. Twice-monthly water sampling, NO₃ ^? injection tests, NO₃ ^? isotopic analysis, filamentous algae mat density, and riparian shading were used to identify processes regulating NO₃ ^? dynamics in the stream over a 12-month period. The urban headwater reach had low NO₃ ^? (0.006–0.2 mg N/L) in the spring through fall, with a minimum uptake length of 900 m, no canopy cover, and high algae mat density. The downstream natural reach (100% canopy cover during the summer and low algae mat density) had nitrate concentrations between 0.6 and 1.2 mg N/L from winter to summer, which decreased during autumn leaf-off. In the urban reach, autotrophic uptake by filamentous green algae is a major NO₃ ^? sink in summer. In the natural reach, the addition of organic matter to the stream at leaf-off led to a decrease in NO₃ ^? concentration followed by an increase in NO₃ ^? concentration in winter as gross primary productivity decreased. This study shows that the balance between autotrophy and heterotrophy in urban streams is variable and depends on an interplay of drivers such as temperature, light, and carbon inputs that are mediated by the riparian ecosystem.     

Selected coccal green algae are not affected by the humic substance Huminfeed<Superscript>®</Superscript> in term of growth or photosynthetic performance

Humic substances (HSs) have been shown to influence growth, photophysiology, and redox homeostasis in phototrophs. However, many ecological studies deliver controversial results indicating inhibitory or stimulating effects depending on the kind of phototrophic organism or type of HSs applied. Here we analyzed the effect of Huminfeed^® (HF), a preparation from leonardite associated with lignite deposits, on growth and photosynthetic performance of three different coccal green algae. Concentrations of HF in the range of 2–20 mg l^?1 dissolved organic carbon (DOC) did neither affect the growth rate nor the light-adapted photosynthetic electron transport. Even photoinhibitory light intensities of 1,600 μmol photons m^?2 s^?1, representing the tenfold of growth light intensity, did not result in a decline of the maximal photosynthetic rate in HF-treated algae. In HF-grown algae, a very subtle decrease by about 10% could be observed in thermoluminescent light emission, a sensitive method to detect changes in photosystem II (PSII) chemistry. However, thermoluminescence (TL) represents only 3% of the light-induced charge separated states in PSII. Hence, rather small changes will not have significant effects on overall photosynthetic performance and growth. Therefore, the physiological effect of HSs on freshwater phototrophs has to be revisited.     

Effects of density on growth rates of four benthic diatoms and variations in biochemical composition associated with growth phase

Diatom cell quantity and their biochemical composition vary among species and are greatly affected by harvest stage or culture conditions. This study compares growth pattern, cell attachment, and biochemical composition of four diatoms suitable for abalone post-larvae: Navicula incerta, Proschkinia sp., Nitzschia sp., and Amphora sp. The four diatoms were grown in F/2 medium at 28.5?±?1.4°C, under 62?±?8?μmol?photons?m^?2?s^?1, at different original inoculating densities (0.05?×?10⁶, 0.10?×?10⁶, and 0.25?×?10⁶?cells?mL^?1) and were harvested in log and stationary phase of growth for biochemical analysis. Total protein, carbohydrate, lipid, and ash composition, as well as fatty acid composition, were determined. All diatoms grew better when inoculated at 0.10?×?10⁶?cell?mL^?1 with Proschkinia sp. reaching the highest cell density of 6.56?×?10⁶?cells?mL^?1 in log phase. Amphora sp. had the highest cell attachment capacity when inoculated at 0.10?×?10⁶?cell?mL^?1 (11,580?cells?mm^?2), whereas N. incerta had the lowest (7,750?cells?mm^?2). Protein and lipid (percent dry weight) contents were generally highest in cells during log phase of growth; Amphora sp. in log phase of growth had the highest lipid content of 9.74% DW, whereas significant differences in carbohydrate between the two growth phases were only observed for Proschkinia sp. Besides, all diatoms had higher energy contents in log phase of growth. There were no significant differences in ash content among the four diatoms. Polyunsaturated fatty acid (PUFA) content ranged from 23.25% to 38.62% of the total fatty acids, and the four diatoms tested were richer in n-3 PUFA than in n-6 PUFA. All the diatoms had significant quantities of 20:5n-3 (EPA) (between 12.69% and 17.68% of TFA), and Proschkinia sp., in log phase of growth, had the highest quantity of arachidonic acid (20:4n-6; ARA). The results highlight the influence of culture conditions and harvest protocols on diatom nutritive value and enabled a preliminary approach towards the selection of novel diatom species.     

Unique picoeukaryotic algal community under multiple environmental stress conditions in a shallow, alkaline pan

Winter phytoplankton communities in the shallow alkaline pans of Hungary are frequently dominated by picoeukaryotes, sometimes in particularly high abundance. In winter 2012, the ice-covered alkaline Zab-szék pan was found to be extraordinarily rich in picoeukaryotic green algae (42–82 × 10⁶ cells ml^?1) despite the simultaneous presence of multiple stressors (low temperature and light intensity with high pH and salinity). The maximum photosynthetic rate of the picoeukaryote community was 1.4 μg C μg chlorophyll a ^?1 h^?1 at 125 μmol m^?2 s^?1. The assimilation rates compared with the available light intensity measured on the field show that the community was considerably light-limited. Estimated areal primary production was 180 mg C m^?2 d^?1. On the basis of the 18S rRNA gene analysis (cloning and DGGE), the community was phylogenetically heterogeneous with several previously undescribed chlorophyte lineages, which indicates the ability of picoeukaryotic communities to maintain high genetic diversity under extreme conditions.