Investigation of biomass and lipids production with Neochloris oleoabundans in photobioreactor

The fresh water microalga Neochloris oleoabundans was investigated for its ability to accumulate lipids and especially triacylglycerols (TAG). A systematic study was conducted, from the determination of the growth medium to its characterization in an airlift photobioreactor. Without nutrient limitation, a maximal biomass areal productivity of 16.5 g m⁻² day⁻¹ was found. Effects of nitrogen starvation to induce lipids accumulation was next investigated. Due to initial N. oleoabundans total lipids high content (23% of dry weight), highest productivity was obtained without mineral limitation with a maximal total lipids productivity of 3.8 g m⁻² day⁻¹. Regarding TAG, an almost similar productivity was found whatever the protocol was: continuous production without mineral limitation (0.5 g m⁻² day⁻¹) or batch production with either sudden or progressive nitrogen deprivation (0.7 g m⁻² day⁻¹). The decrease in growth rate reduces the benefit of the important lipids and TAG accumulation as obtained in nitrogen starvation (37% and 18% of dry weight, respectively).     

Utilization of the cyanobacteria Anabaena sp. ATCC 33047 in CO2 removal processes

In this paper the utilization of the cyanobacteria Anabaena sp. in carbon dioxide removal processes is evaluated. For this, continuous cultures of this strain were performed at different dilution rates; alternatives for the recovery of the organic matter produced being also studied. A maximum CO₂ fixation rate of 1.45 g CO₂ L⁻¹ day⁻¹ was measured experimentally, but it can be increased up to 3.0 g CO₂ L⁻¹ day⁻¹ outdoors. The CO₂ is mainly transformed into exopolysaccharides, biomass representing one third of the total organic matter produced. Organic matter can be recovered by sedimentation with efficiencies higher than 90%, the velocity of sedimentation being 2 · 10⁻⁴ s⁻¹. The major compounds were carbohydrates and proteins with productivities of 0.70 and 0.12 g L⁻¹ day⁻¹, respectively. The behaviour of the cultures of Anabaena sp. has been modelized, also the characteristics parameters requested to design separation units being reported. Finally, to valorizate the organic matter as biofertilizers and biofuels is proposed.     

Production and recovery of recombinant protease inhibitor α1-antitrypsin

Human α₁-antitrypsin (AAT) was produced in the recombinant yeast Saccharomyces cerevisiae ATCC 20699 grown in batch and fed-batch culture. The final biomass concentration and antitrypsin concentration attained were 55 g·L⁻¹ and 1.23 g·L⁻¹, respectively, in the fed-batch. The maximum productivities of biomass and antitrypsin were 1.6 and > 0.04 g L⁻¹h⁻¹, respectively, or substantially greater than the highest productivity values reported in the past. For recovering the antitrypsin, the cell slurry was concentrated 4-fold (231 g·L⁻¹ biomass, 122 min of processing) by cross-flow microfiltration and the cells were disrupted by bead milling (3 passes of 3 min total retention time). The cell homogenate was treated with aluminum chloride or PBS (pH 7) to aid separation of the cell debris by flocculation and sedimentation. The clarified cell homogenate was subjected to ammonium sulfate fractionation to precipitate the recombinant antitrypsin. The AAT precipitated at 45–75% saturation of ammonium sulfate, depending on the age of the homogenate. The crude AAT in the homogenate degraded at room temperature (25°C), with a zero order deactivation rate of 1.815 × 10⁻³ ± 3.43 × 10⁻⁴ g AAT L⁻¹h⁻¹.     

The effect of environmental factors on the growth rate of Karenia brevis (Davis) G. Hansen and Moestrup

The red tide dinoflagellate Karenia brevis (Davis) G. Hansen and Moestrup is noted for causing mass mortalities of marine organisms in the Gulf of Mexico. Most research has focused on culture isolates from the eastern Gulf of Mexico. In this investigation, we examine the effects of light, temperature and salinity on the growth rate of K. brevis from the western Gulf of Mexico. Growth rates of K. brevis were determined under various combinations of irradiance (19, 31, 52, 67, and 123 μmol m⁻² s⁻¹), salinity (25, 30, 35, 40 and 45), and temperature (15, 20, 25, and 30 °C). Maximum growth rates varied from 0.17 to 0.36 div day⁻¹ with exponential growth rates increasing with increasing irradiance. Little or no growth was supported at 19 μmol photons m⁻² s⁻¹ for any experiment. Maximum growth rates at 15 °C were much lower than at other temperatures. Maximum growth rates of the Texas clone (SP3) fell within the range of Florida clones reported in the literature (0.17–0.36 div day⁻¹ versus 0.2–1.0 div day⁻¹). The Texas clone SP3 had a very similar light saturation point compared to that of a Florida isolate (Wilson's clone) (67 μmol m⁻² s⁻¹ versus 65 μmol m⁻² s⁻¹), and light compensation (20–30 μmol m⁻² s⁻¹1). The upper and lower salinity tolerance of the Texas clone was similar than that of some Florida clones (45 versus 46 and 25 versus 22.5, respectively). In our study, the Texas clone had the same temperature tolerance reported for Florida clones (15–30 °C). While individual clones can vary considerably in maximum growth rates, our results indicate only minor differences exist between the Texas and Florida strains of K. brevis in their temperature and salinity tolerance for growth. While the literature notes lower salinity occurrences of K. brevis in nearby Louisiana, our isolate from the southern Texas coast has the higher salinity requirements typical of K. brevis in the eastern Gulf of Mexico.     

Spacer elongation and plagiotropic growth are the primary clonal strategies used by Vallisneria spiralis to acclimate to sedimentation

The aim of this study is to identify how submerged macrophyte Vallisneria spiralis acclimate to sedimentation by investigating the growth, biomass allocation and clonal characteristics in a greenhouse experiment of 30 days. Experimental treatments combined two sediment types (mud and sand) with four sedimentation depths (0, 2, 4 and 8 cm) in a factorial design. Biomass accumulation (0.98–1.33 versus 0.36 g per plant) and relative growth rate (RGR, 0.082–0.093 versus 0.046 g g⁻¹ day⁻¹) decreased only in the 8 cm sand treatment. Neither sedimentation depth nor sediment type influenced biomass allocation. The ratio of spacer length to biomass was significantly higher in the 8 cm sand (20.4 cm g⁻¹) than in other treatments (6.0–8.5 cm g⁻¹). Branching angles and the depths between ramet basal and sediment surface were only affected by sedimentation depth. Clonal ramets developed nearly vertical branching angles (ranged from 78° to 101°) in the 0 cm sedimentation treatment, but the angles of treated plants decreased at the initial 3–5 ramets (ranged from 68° to 78° at the first ramet level), then remained a relatively constant value (about 90°) in the following spacers. These data indicate that plagiotropic stolons were formed to project the ramets to sediment surface and to escape sedimentation stress primarily by elongating spacer length and decreasing branching angle, rather than by adjusting biomass allocation.     

Effects of growth conditions on exopolysaccharide production by Cyanospira capsulata

Batch cultures of Cyanospira capsulata, a heterocystous cyanobacterium possessing a thick polysaccharidic capsule, were characterized by increasing viscosity owing to the continuous release of a soluble polysaccharide (EPS) into the culture medium. Both capsulated trichomes and solubilized EPS contributed to the flow properties of whole cultures. A typical pseudoplastic behaviour, the more marked the more aged were the cultures, was evidenced.The production of EPS was investigated under different growth conditions by changing some nutritional and physical parameters known to affect the synthesis of exopolysaccharides in algae and cyanobacteria. Among the factors tested (Ca²⁺, Mg²⁺ or PO₄⁻⁴ deficiencies, salinity and pH) only Mg²⁺ shortage caused a significant enhancement of the EPS production. Under continuous illumination in open ponds, the EPS productivity of batch cultures on standard mineral medium was about 5·8 g m⁻² day⁻¹, whereas under Mg²⁺ deficiency with a consequent increase of the cultures' viscosity     

Effects of dwarf bamboo (Fargesia denudata) density on biomass, carbon and nutrient distribution pattern

Dense dwarf bamboo population is a structurally and functionally important component in many subalpine forest systems. To characterize the effects of stem density on biomass, carbon and majority nutrients (N, P, K, Ca and Mg) distribution pattern, three dwarf bamboo (Fargesia denudata) populations with different stem densities (Dh with 220 ± 11 stems m^?2, Dm with 140 ± 7 stems m^?2, and Dl with 80 ± 4 stems m^?2, respectively) were selected beneath a bamboo-fir (Picea purpurea) forest in Wanglang National Nature Reserve, Sichuan, China. Leaf, branch, rhizome, root and total biomass of dwarf bamboo increased with the increase of stem density, while carbon and nutrient concentrations in bamboo components decreased. Percentages of below-ground biomass and element stocks to total biomass and stocks decreased with the increase of stem density, whereas above-ground biomass and element stocks exhibited the opposite tendency. Moreover, more above-ground biomass and elements were allocated to higher part in the higher density population. In addition, percentages of culm biomass, above-ground biomass and element stocks below 100 cm culm height (H₁₀₀) increased with the increase of stem density, while percentages of branch and leaf biomass below H₁₀₀ decreased. Pearson’s correlation analyses revealed that root biomass, above-ground biomass, below-ground biomass and total biomass significantly correlated to leaf biomass in H_100?200 and total leaf biomass within high density population, while they significantly correlated to leaf biomass in H_50?150 within low density population. The results suggested that dwarf bamboo performed an efficient adaptive strategy to favor limited resources by altering biomass, carbon and nutrients distribution pattern in the dense population.     

Export production of coccolithophores in an upwelling region: Results from San Pedro Basin, Southern California Borderlands

A seven month-long time series sediment trap project was carried out in San Pedro Basin (Southern California Borderlands) in order to evaluate the response of calcareous nannoplankton to seasonal hydrographic changes. This region is periodically influenced by upwelling, particularly during the spring and early summer. The highest fluxes of both whole coccospheres and individual coccoliths occurred during winter (January-February), a period when the fluxes of diatoms and planktic foraminifera were low. The highest coccolithophore fluxes were recorded in the mid-February with 860 × 10⁶ coccoliths m⁻² day⁻¹, 8 × 10⁶ whole coccospheres m⁻² day⁻¹, and 80 mg of coccolith carbonate m⁻² day⁻¹. Coccolith carbonate fluxes in January and February account for most of the total carbonate fluxes measured during this period. The season of maximum coccolithophore production in this region (winter) is correlated with weak stratification of the upper water column, low total primary production, low nutrient contents, and low temperatures.Emiliania huxleyi and Florisphaera profunda are the two most abundant species in this region. While E. huxleyi displays no distinct seasonal changes in flux, F. profunda shows a clear preference for cold, low nutrient water conditions and low light levels. Helicosphaera spp. flux is positively correlated to the total coccosphere fluxes and is indicative of high coccolithophore productivity.     

Growth of dinoflagellates, Ceratium furca and Ceratium fusus in Sagami Bay, Japan: The role of temperature, light intensity and photoperiod

Seasonal changes of field populations and growth rates of two dinoflagellates, Ceratium furca and Ceratium fusus, were examined in the temperate coastal water of Sagami Bay, Japan. Weekly field sampling was conducted from August 2002 to August 2003, and laboratory experiments were also carried out to investigate effects of temperature, irradiance and photoperiod on the growth rates of these two Ceratium species. In the field, the abundances of both species increased significantly from April to August 2003, were gradually decreased from November 2002 and were not observed in January 2003. C. fusus was able to increase at lower temperatures in February 2003 compared to C. furca. In the laboratory, the two species did not grow at <10 °C or >32 °C. The highest specific growth rate of C. furca was 0.72 d⁻¹ at 24 °C and 600 μmol m⁻² s⁻¹. Optimum growth rates (>0.4 d⁻¹) of C. furca were observed at temperatures from 18 to 28 °C and at irradiances from 216 to 796 μmol m⁻² s⁻¹. The highest growth rate of C. fusus was 0.56 d⁻¹ at 26 °C and 216 μmol m⁻² s⁻¹. Optimum growth rates of C. fusus were observed at the same irradiance rage of C. furca, whereas optimum temperature range was narrower (26–28 °C). The growth curves of both species indicated saturation of the growth rates when light intensity was above 216 μmol m⁻² s⁻¹, and did not show photoinhibition at irradiances up to 796 μmol m⁻² s⁻¹. The specific growth rates of both Ceratium species were clearly decreased at L:D = 10:14 relative to those at L:D = 14:10 and L:D = 12:12. The present study indicates the two Ceratium species can adapt to a wide range of temperature and irradiance.     

Annual cycle of CO2 exchange over a reed (Phragmites australis) wetland in Northeast China

Net ecosystem exchange of CO₂ (NEE) was measured during 2005 using the eddy covariance (EC) technique over a reed (Phragmites australis (Cav.) Trin. ex Steud.) wetland in Northeast China (121°54′E, 41°08′N). Diurnal NEE patterns varied markedly among months. Outside the growing season, NEE lacked a diurnal pattern and it fluctuated above zero with an average value of 0.07 mg CO₂ m⁻² s⁻¹ resulting from soil microbial activity. During the growing season, NEE showed a distinct V-like diel course, and the mean daily NEE was −7.48 ± 2.74 g CO₂ m⁻² day⁻¹, ranging from −13.58 g CO₂ m⁻² day⁻¹ (July) to −0.10 g CO₂ m⁻² day⁻¹ (October). An annual cycle was also apparent, with CO₂ uptake increasing rapidly in May, peaking in July, and decreasing from August. Monthly cumulative NEE ranged from −115 ± 24 g C m⁻² month⁻¹ (the reed wetland was a CO₂ sink) in July to 75 ± 16 g C m⁻² month⁻¹ (CO₂ source) in November. The annual CO₂ balance suggests a net uptake of −65 ± 14 g C m⁻² year⁻¹, mainly due to the gains in June and July. Cumulative CO₂ emission during the non-growing season was 327 g C m⁻², much greater than the absolute value of the annual CO₂ balance, which proves the importance of the wintertime CO₂ efflux at the study site. The ratio of ecosystem respiration (R_eco) to gross primary productivity (GPP) for this reed ecosystem was 0.95, indicating that 95% of plant assimilation was consumed by the reed plant or supported the activities of heterotrophs in the soil. Daytime NEE values during the growing season were closely related to photosynthetically active radiation (PAR) (r² > 0.63, p < 0.01). Both maximum ecosystem photosynthesis rate (A_max) and apparent quantum yield (α) were season-dependent, and reached their peak values in July (1.28 ± 0.11 mg CO₂ m⁻² s⁻¹, 0.098 ± 0.027 μmol CO₂ μmol⁻¹ photon, respectively), corresponding to the observed maximum NEE in July. Ecosystem respiration (R_eco) relied on temperature and soil water content, and the mean value of Q₁₀ was about 2.4 with monthly variation ranging from 1.8 to 4.1 during 2005. Annual methane emission from this reed ecosystem was estimated to be about 3 g C m⁻² year⁻¹, and about 5% of the net carbon fixed by the reed wetland was released to the atmosphere as CH₄.     

Bottlenecks to water transport in Quercus robur L.: the abscission zone and its physiological consequences

Cladoptosis, the abscission of twigs, is the main mechanism of changes in crown structure in senescing pedunculate oak (Quercus robur L.). We tested the hypotheses that abscission zones in nodes of old pedunculate oak trees reduce leaf-specific hydraulic conductance of shoots and thereby limit the stomatal conductance and assimilation.Hydraulic conductance and leaf-specific hydraulic conductance, measured with a high pressure flowmeter in 0.5–1.5 m long shoots, were significantly lower in shoots of low vigour compared to vigorous growing shoots in a 165-years-old stand in the southeast of Germany. Two types of bottlenecks to water transport could be identified in shoots of old oak trees, namely nodes and abscission zones. In young twigs, vessel diameter and vessel density in nodes with abscission zones were significantly reduced compared with internodes. In nodes without abscission zones, vessel density was significantly reduced. The reduction of hydraulic conductance was especially severe in the smallest and youngest shoots with diameters less than 2 mm. Internodes of 1–5 mm sapwood diameter had an average hydraulic conductance of 7.13×10⁻⁶±0.2×10⁻⁶ kg s⁻¹ m⁻¹ MPa⁻¹, compared to 4.54×10⁻⁶±0.3×10⁻⁶ kg s⁻¹ m⁻¹ MPa⁻¹ in those with nodes.Maximum stomatal conductance and maximum net assimilation rate increased significantly with hydraulic conductance and leaf-specific hydraulic conductance. Maximum rate of net photosynthesis A_max of the most vigorous shoots (VC0) (7.34±0.55 μmol m⁻² s⁻¹) was significantly higher (P<0.001) than in shoots of other vigour classes (5.97±0.28 μmol m⁻² s⁻¹). Our data support the hypothesis that the changes in shoot and consequently crown architecture that are observed in ageing and declining trees can limit photosynthesis by reducing shoot hydraulic conductance. Abscission zones increase the hydraulic disadvantage of less vigorous compared to vigorously growing twigs. Cladoptosis might serve as a mechanism of selection between twigs of different efficiency.     

Aquatic metabolism as an indicator of the ecological effects of hydrologic pulsing in flow-through wetlands

Water column metabolism is a major component in the functioning of wetland ecosystems and can be used as an indicator of ecosystem health. The effect of hydrologic pulsing on water column metabolism was studied with 2 year's field data and a validated model for two 1-ha created riparian wetlands in Columbus, OH, USA. Aquatic gross primary productivity (GPP) was measured during hydrologic pulses the first week of April, May and June of 2004 and compared to GPP during steady flow-conditions in April, May and June 2005. Pulses reduced diurnal variation of water temperature, pH and dissolved oxygen, and negatively affected GPP rates. Mean GPP measured during hydrologic flood pulses was 5.4 ± 2.6 kcal m⁻² day⁻¹, significantly lower than that measured for comparable months with steady-flow hydrology (10.8 ± 3.3 kcal m⁻² day⁻¹). Solar-normalized productivity values of 0.08 ± 0.01% of solar energy during pulses and 0.2 ± 0.02% for steady-flow conditions were also significantly different. Different hyperbolic curves of optimum productivity with water temperature were seen for pulsing and steady-flow conditions. A simulation model with hydrology, metabolism, and dissolved oxygen sub models was calibrated with 2005 steady-flow year data and validated with 2004 pulse year data. Results from both the field study and model simulations suggest that there was a threshold hydraulic inflow rate between 30 and 50 cm day⁻¹ where aquatic metabolism became negatively affected by flow.     

Influence of soft rush (Juncus effusus) on phosphorus flux in grazed seasonal wetlands

Livestock significantly affect wetland soils and vegetation but their impacts on wetland nutrient dynamics are poorly understood. We set up a full factorial laboratory experiment to assess the effects of Juncus effusus, grazing exclusion, and flooding on P flux from intact cores collected from seasonal wetlands in cattle pastures in south Florida. We collected intact cores from Juncus tussocks and plant interspaces inside and outside 4-year grazing exclosures in five replicate wetlands. We incubated the cores for 50 days under continuous flooding or weekly 1-day flooding cycles and measured P concentrations in surface and pore water. Grazing exclosures had less Juncus (17%) and bare ground (2%) than adjacent grazed areas (Juncus, 48%; bare ground, 12%), but did not affect P fluxes. Initial fluxes of soluble reactive P (SRP) were much higher in cores with Juncus (242 ± 153 mg P m⁻² day⁻¹) than without Juncus (14 ± 20 mg P m⁻² day⁻¹). In weekly flooded cores P fluxes fell to 19.7 ± 13.4 mg P m⁻² day⁻¹ in cores with and 2.7 ± 2.6 in cores without Juncus. The strong effect of Juncus on P flux was an indirect effect of cattle grazing, but 4 years of grazing exclusion did not have a significant effect on P fluxes.     

A novel photobioreactor with transparent rectangular chambers for cultivation of microalgae

An open tank photobioreactor containing transparent rectangular chambers (TRCs) was developed to improve the photosynthetic efficiency of microalgal cultivation. The TRCs, made of transparent acrylic, conducted light deep into the photobioreactor, especially at high cell concentrations. The average irradiance, I_av, was calculated by Lambert–Beer's law, and was used to determine the light conditions in the cultivation system. The photobioreactor provided large areas of illumination that improved the effective utilization of light energy for microalgae growth and created a good artificial environment for a high rate of cell growth, even at low I_av. The biomass concentration of Chlorella sp. reached 3.745 g L⁻¹ on the 13th day, with biomass productivity of 0.340 g L⁻¹ d⁻¹. The total biomass obtained was 56% more than that of similar culture systems without TRCs.     

Benthic microalgae in coral reef sediments of the southern Great Barrier Reef,Australia

The abundance and productivity of benthic microalgae in coral reef sediments are poorly known compared with other, more conspicuous (e.g. coral zooxanthellae, macroalgae) primary producers of coral reef habitats. A survey of the distribution, biomass, and productivity of benthic microalgae on a platform reef flat and in a cross-shelf transect in the southern Great Barrier Reef indicated that benthic microalgae are ubiquitous, abundant (up to 995.0 mg chlorophyll (chl) a m^–2), and productive (up to 110 mg O₂ m^–2 h^–1) components of the reef ecosystem. Concentrations of benthic microalgae, expressed as chlorophyll a per surface area, were approximately 100-fold greater than the integrated water column concentrations of microalgae throughout the region. Benthic microalgal biomass was greater on the shallow water platform reef than in the deeper waters of the cross-shelf transect. In both areas the benthic microalgal communities had a similar composition, dominated by pennate diatoms, dinoflagellates, and cyanobacteria. Benthic microalgal populations were potentially nutrient-limited, based on responses to nitrogen and phosphorus enrichments in short-term (7-day) microcosm experiments. Benthic microalgal productivity, measured by O₂ evolution, indicated productive communities responsive to light and nutrient availability. The benthic microalgal concentrations observed (92–995 mg chl a m^–2) were high relative to other reports, particularly compared with temperate regions. This abundance of productive plants in both reef and shelf sediments in the southern Great Barrier Reef suggests that benthic microalgae are key components of coral reef ecosystems.Communicated by Environmental Editor, B.C. Hatcher     

Periphyton as a potential phosphorus sink in the Everglades Nutrient Removal Project

Phosphorus uptake and release by periphyton mats were quantified in the Everglades Nutrient Removal Project (ENRP) to evaluate the potential for periphyton P removal. Short-term P uptake rates were determined by incubating cyanobacteria (Oscillatoria princeps and Shizothrix calcicola) and Chlorophycean (primarily Rhizoclonium spp.) algal mat samples for 0.5–2 h under ambient conditions in BOD bottles spiked with soluble reactive P (SRP). Cyanobacterial mats removed P more than twice as fast (80–164 μg P h⁻¹ g⁻¹ AFDM) as Chlorophycean mats (33–61 μg P h⁻¹ g⁻¹ AFDM) during these incubations. In a longer term study, fiberglass cylinders were used to enclose 1.8 m² plots within the wetland and were dosed weekly for 7 weeks with: (1) no nutrients; (2) SRP (0.25 g P m⁻² week⁻¹); or (3) SRP plus nitrate (0.42 g N m⁻² week⁻¹) and ammonium (0.83 g N m⁻² week⁻¹). Phosphorus uptake rates by this periphyton assemblage, which was dominated by the chlorophytes Stigeoclonium spp. and Oedogonium spp., were measured weekly and were similar among nutrient treatments on most dates, indicating that the algal storage compartment for P was not saturated despite repeated P additions. Decomposition rates and P loss by cyanobacteria and Chlorophycean mats were determined by measuring biomass loss and SRP release in darkened BOD bottles over 28–42 day periods under anaerobic and aerobic conditions. First-order aerobic and anaerobic decomposition rates for cyanobacterial mats (k = 0.1095 and 0.1408 day⁻¹, respectively) were 4–20-fold higher than rates for Chlorophycean mats (k = 0.0066 and 0.0250 day⁻¹, respectively) and cyanobacteria released considerably more P back to the water column. Our findings suggest that periphyton can be an important short-term sink for P in treatment wetlands and that retention is strongly affected by the taxonomic composition of the periphyton assemblage.     

Nitrate and phosphate removal by<Emphasis Type="Italic"> Spirulina platensis</Emphasis>

The cyanobacterium Spirulina platensis was used to verify the possibility of employing microalgal biomass to reduce the contents of nitrate and phosphate in wastewaters. Batch tests were carried out in 0.5 dm³ Erlenmeyer flasks under conditions of light limitation (40 mol quanta m^–2 s^–1) at a starting biomass level of 0.50 g/dm³ and varying temperature in the range 23–40°C. In this way, the best temperature for the growth of this microalga (30°C) was determined and the related thermodynamic parameters were estimated. All removed nitrate was used for biomass growth (biotic removal), whereas phosphate appeared to be removed mainly by chemical precipitation (abiotic removal). The best results in terms of specific and volumetric growth rates ( =0.044 day^–1, Q _x =33.2 mg dm^–3 day^–1) as well as volumetric rate and final yield of nitrogen removal ( =3.26 mg dm^–3 day^–1, =0.739) were obtained at 30°C, whereas phosphorus was more effectively removed at a lower temperature. In order to simulate full-scale studies, batch tests of nitrate and phosphate removal were also performed in 5.0 dm³ vessels (mini-ponds) at the optimum temperature (30°C) but increasing the photon fluence rate to 80 mol quanta m^–2 s^–1 and varying the initial biomass concentration from 0.25 to 0.86 g/dm³. These additional tests demonstrated that an increase in the inoculum level up to 0.75 g/dm³ enhanced both NO₃ ^– and PO₄ ^3– removal, confirming a strict dependence of these processes on biomass activity. In addition, the larger surface area of the ponds and the higher light intensity improved removal yields and kinetics compared to the flasks, particularly concerning phosphorus removal ( =0.032–0.050 day^–1, Q _x =34.7–42.4 mg dm^–3 day^–1, =3.24–4.06 mg dm^–3 day^–1, =0.750–0.879, =0.312–0.623 mg dm^–3 day^–1, and =0.224–0.440).     

The potential role of climate in the distribution and zonation of the introduced seagrass Zostera japonica in North America

The current distribution of the introduced seagrass Zostera japonica is restricted to the mid- to upper intertidal zone in the coastal Pacific Northwest region of North America. The climate in this region is cool and wet, becoming hotter and dryer with increasing distance southward. Since temperature is likely to be an important factor affecting distribution of this species, growth of two populations located near the northern and southern limits of its established range along the Pacific Coast of North America were measured in an experimental setting across a range of temperatures typical of those in the field during the growing season (10, 20, and 30 °C). The effects of temperature and population were both significant. Leaf elongation, growth, and areal productivity rates of the northern population were consistently lower than those of the southern population. Across the range of temperatures, mean leaf elongation rates ranged from 0.47 to 1.40 cm² shoot⁻¹ d⁻¹; mean growth rates ranged from 0.19 to 0.52 mg dry wt shoot⁻¹ d⁻¹. Mean areal productivity ranged from 0.54 to 1.92 g dry wt m⁻² d⁻¹. Maximum rates of leaf elongation, growth, and areal productivity for both populations were observed at 20 °C. However, leaf elongation, growth, and areal productivity of the northern population declined markedly at 30 °C, whereas no comparable declines were observed for the southern population. This suggests that Z. japonica populations near the southern limits of its established range may be better adapted to warmer temperatures than populations near the northern range limits and further range extensions southward along the California coast may be likely. These differences could be important in predicting the outcome of competitive interactions between native and introduced seagrass species, and in determining future patterns of distribution and zonation of Pacific Coast seagrasses.     

Seasonal differences in activity of perch (Perca flavescens) gill Na/K ATPase

We measured Na⁺/K⁺ ATPase activity in homogenates of gill tissue prepared from field caught, winter and summer acclimatized yellow perch, Perca flavescens. Water temperatures were 2–4°C in winter and 19–22°C in summer. Na⁺/K⁺ ATPase activity was measured at 8, 17, 25, and 37°C. V_max values for winter fish increased from 0.48±0.07 μmol P mg⁻¹ protein h⁻¹ at 8°C to 7.21±0.79 μmol P mg⁻¹ protein h⁻¹ at 37°C. In summer fish it ranged from 0.46±0.08 (8°C) to 3.86±0.50 (37°C) μmol P mg⁻¹ protein h⁻¹. The K_m for ATP and for Na⁺ at 8°C was ≈1.6 and 10 mM, respectively and did not vary significantly with assay temperature in homogenates from summer fish. The activation energy for Na⁺/K⁺ ATPase from summer fish was 10 309 (μmol P mg⁻¹ h⁻¹) K⁻¹. In winter fish, the K_m for ATP and Na⁺ increased from 0.59±0.08 mM and 9.56±1.18 mM at 8°C to 1.49±0.11 and 17.88±2.64 mM at 17°C. The K_m values for ATP and Na did not vary from 17 to 37°C. A single activation energy could not be calculated for Na/K ATPase from winter fish. The observed differences in enzyme activities and affinities could be due to seasonal changes in membrane lipids, differences in the amount of enzyme, or changes in isozyme expression.