Nitrogen Utilization and Toxin Production by Two Diatoms of the Pseudo‐nitzschia pseudodelicatissima Complex: P. cuspidata and P. fryxelliana

The toxigenic diatom Pseudo‐nitzschia cuspidata, isolated from the U.S. Pacific Northwest, was examined in unialgal batch cultures to evaluate domoic acid (DA) toxicity and growth as a function of light, N substrate, and growth phase. Experiments conducted at saturating (120 μmol photons · m^?2 · s^?1) and subsaturating (40 μmol photons · m^?2 · s^?1) photosynthetic photon flux density (PPFD), demonstrate that P. cuspidata grows significantly faster at the higher PPFD on all three N substrates tested [nitrate (NO₃^?), ammonium (NH₄⁺), and urea], but neither cellular toxicity nor exponential growth rates were strongly associated with one N source over the other at high PPFD. However, at the lower PPFD, the exponential growth rates were approximately halved, and the cells were significantly more toxic regardless of N substrate. Urea supported significantly faster growth rates, and cellular toxicity varied as a function of N substrate with NO₃^?‐supported cells being significantly more toxic than both NH₄⁺‐ and urea‐supported cells at the low PPFD. Kinetic uptake parameters were determined for another member of the P. pseudodelicatissima complex, P. fryxelliana. After growth of these cells on NO₃^? they exhibited maximum specific uptake rates (V_max) of 22.7, 29.9, 8.98 × 10^?3 · h^?1, half‐saturation constants (K_s) of 1.34, 2.14, 0.28 μg‐at N · L^?1, and affinity values (α) of 17.0, 14.7, 32.5 × 10^?3 · h^?1/(μg‐at N · L^?1) for NO₃^?, NH₄⁺ and urea, respectively. These labo‐ratory results demonstrate the capability of P. cuspidata to grow and produce DA on both oxidized and reduced N substrates during both exponential and stationary growth phases, and the uptake kinetic results for the pseudo‐cryptic species, P. fryxelliana suggest that reduced N sources from coastal runoff could be important for maintenance of these small pennate diatoms in U.S. west coast blooms, especially during times of low ambient N concentrations.     

Variation in growth rate,carbon assimilation,and photosynthetic efficiency in response to nitrogen source and concentration in phytoplankton isolated from upper San Francisco Bay

Six species of phytoplankton recently isolated from upper San Francisco Bay were tested for their sensitivity to growth inhibition by ammonium (NH₄⁺), and for differences in growth rates according to inorganic nitrogen (N) growth source. The quantum yield of photosystem II (F_v/F_m) was a sensitive indicator of NH₄⁺ toxicity, manifested by a suppression of F_v/F_m in a dose‐dependent manner. Two chlorophytes were the least sensitive to NH₄⁺ inhibition, at concentrations of >3,000 μmoles NH₄⁺ · L^?1, followed by two estuarine diatoms that were sensitive at concentrations >1,000 μmoles NH₄⁺ · L^?1, followed lastly by two freshwater diatoms that were sensitive at concentrations between 200 and 500 μmoles NH₄⁺ · L^?1. At non‐inhibiting concentrations of NH₄⁺, the freshwater diatom species grew fastest, followed by the estuarine diatoms, while the chlorophytes grew slowest. Variations in growth rates with N source did not follow taxonomic divisions. Of the two chlorophytes, one grew significantly faster on nitrate (NO₃^?), whereas the other grew significantly faster on NH₄⁺. All four diatoms tested grew faster on NH₄⁺ compared with NO₃^?. We showed that in cases where growth rates were faster on NH₄⁺ than they were on NO₃^?, the difference was not larger for chlorophytes compared with diatoms. This holds true for comparisons across a number of culture investigations suggesting that diatoms as a group will not be at a competitive disadvantage under natural conditions when NH₄⁺ dominates the total N pool and they will also not have a growth advantage when NO₃^? is dominant, as long as N concentrations are sufficient.     

Kinetic Modeling for Simultaneous Organic Carbon Oxidation,Nitrification, and Denitrification of Abattoir Wastewater in Sequencing Batch Reactor

A laboratory-scale study was conducted in a 20.0-L sequencing batch reactor (SBR) to explore the feasibility of simultaneous removal of organic carbon and nitrogen from abattoir wastewater. The reactor was operated under three different combinations of aerobic-anoxic sequence, viz., (4+4), (5+3), and (5+4) h of total react period, with influent soluble chemical oxygen demand (SCOD) and ammonia nitrogen (NH₄⁺-N) level of 2200 ± 50 and 125 ± 5 mg L^?1, respectively. In (5+4) h cycle, a maximum 90.27% of ammonia reduction corresponding to initial NH₄⁺-N value of 122.25 mg L^?1 and 91.36% of organic carbon removal corresponding to initial SCOD value of 2215.25 mg L^?1 have been achieved, respectively. The biokinetic parameters such as yield coefficient (Y), endogenous decay constant (k_d), and half-velocity constant (K_s) were also determined to improve the design and operation of package effluent treatment plants comprising SBR units. The specific denitrification rate (q_DN) during anoxic condition was estimated as 6.135 mg N/g mixed liquor volatile suspended solid (MLVSS)·h on 4-h average contact period. The value of Y, k_d and K_s for carbon oxidation and nitrification were found to be in the range of 0.6225–0.6952 mg VSS/mg SCOD, 0.0481–0.0588 day^?1, and 306.56–320.51 mg L^?1, and 0.2461–0.2541 mg VSS/mg NH₄⁺-N, 0.0324–0.0565 day^?1, and 38.28–50.08 mg L^?1, respectively, for different combinations of react periods.     

Dry deposition of ammonia gas drives species change faster than wet deposition of ammonium ions: evidence from a long‐term field manipulation

Although the effects of atmospheric nitrogen deposition on species composition are relatively well known, the roles of the different forms of nitrogen, in particular gaseous ammonia (NH₃), have not been tested in the field. Since 2002, we have manipulated the form of N deposition to an ombrotrophic bog, Whim, on deep peat in southern Scotland, with low ambient N (wet + dry = 8 kg N ha^?1 yr^?1) and S (4 kg S ha^?1 yr^?1) deposition. A gradient of ammonia (NH₃, dry N), from 70 kg N ha^?1 yr^?1 down to background, 3–4 kg N ha^?1 yr^?1 was generated by free air release. Wet ammonium (NH₄⁺, wet N) was provided to replicate plots in a fine rainwater spray (NH₄Cl at +8, +24, +56 kg N ha^?1 yr^?1). Automated treatments are coupled to meteorological conditions, in a globally unique, field experiment. Ammonia concentrations were converted to NH₃‐N deposition (kg N ha^?1) using a site/vegetation specific parameterization. Within 3 years, exposure to relatively modest deposition of NH₃, 20–56 kg NH₃‐N ha^?1 yr^?1 led to dramatic reductions in species cover, with almost total loss of Calluna vulgaris, Sphagnum capillifolium and Cladonia portentosa. These effects appear to result from direct foliar uptake and interaction with abiotic and biotic stresses, rather than via effects on the soil. Additional wet N by contrast, significantly increased Calluna cover after 5 years at the 56 kg N dose, but reduced cover of Sphagnum and Cladonia. Cover reductions caused by wet N were significantly different from and much smaller than those caused by equivalent dry N doses. The effects of gaseous NH₃ described here, highlight the potential for ammonia to destroy acid heathland and peat bog ecosystems. Separating the effects of gaseous ammonia and wet ammonium deposition, for a peat bog, has significant implications for regulatory bodies and conservation agencies.     

AMMONIUM AND UV RADIATION STIMULATE THE ACCUMULATION OF MYCOSPORINE‐LIKE AMINO ACIDS IN PORPHYRA COLUMBINA (RHODOPHYTA) FROM PATAGONIA,ARGENTINA1

The combined effects of ammonium concentration and UV radiation on the red alga Porphyra columbina (Montagne) from the Patagonian coast (Chubut, Argentina) was determined using short‐term (less than a week) experimentation. Discs of P. columbina were incubated with three ammonium concentrations (0, 50, and 300 μM NH₄Cl) in anilluminated chamber (PAR=300 μmol photons·m^?2·s^?1, UVA=15 W·m^?2, UVB=0.7 W·m^?2) at 15°C. Algae incubated at 300 μM ammonium showed a significant increase (P<0.05) in the concentration of mycosporine‐like amino acids (MAAs) compared with the initial value or with the other ammonium treatments. The increase of MAAs was, however, a function of the quality of irradiance received, with a higher increase in samples exposed to UVA compared with UVB (29% and 5% increase, respectively). However, UVB radiation was more effective in inducing MAA synthesis per unit energy received by the algae. Samples exposed to PAR only had an intermediate increase in MAA concentration of 16%. Chl a concentration decreased through the incubation with the greatest decrease at high ammonium concentration. Phycobiliprotein (BP) decreased through time with the smallest decrease occurring at high ammonium concentration. Photoinhibition (as a decrease of optimal quantum yield) was significantly greater under nitrogen‐deprived conditions than that under replete ammonium levels. Maximal gross photosynthesis (GP_max), as oxygen evolution, and maximal electron transport rate (ETR_max), as chl fluorescence, increased with the ammonium concentration. Positive relationships between maximal GP or ETR and pigment ratios (BP/chl a and MAAs/chl a) and negative relationships with chl a concentration were found.     

INTERACTIONS BETWEEN LIGHT,NH4+, AND CO2 IN BUOYANCY REGULATION OF ANABAENA FLOS-AQUAE (CYANOPHYCEAE)1

Buoyancy of the gas-vacuolate alga Anabaena flosaquae Brébisson was measured under various levels of light, NH₄⁺, and CO₂. At high irradiance (50 μE · m^?2·^?1) the alga was non-buoyant regardless of the availability of CO₂ and NH₄⁺. At low irradiance (≤10 μE · m ^?2· s^?1) buoyancy was controlled by the availability of NH₄⁺ and CO₂. When NH₄⁺ was abundant, algal buoyancy was high over a wide range of CO₂ concentrations. In the absence of NH₄⁺, algal buoyancy was reduced at high CO₂ concentrations, however as the CO₂ concentration declined below about 5 μmol · L^?1, algal buoyancy increased. These results help explain why gas vacuolate, nitrogen-fixing blue-green algae often form surface blooms in eutrophic lakes.     

Production rate estimation of mycosporine‐like amino acids in two Arctic melt ponds by stable isotope probing with NAH13CO3

The net carbon uptake rate and net production rate of mycosporine‐like amino acids (MAAs) were measured in phytoplankton from 2 different melt ponds (MPs; closed and open type pond) in the western Arctic Ocean using a ¹³C stable isotope tracer technique. The Research Vessel Araon visited ice‐covered western‐central basins situated at 82°N and 173°E in the summer of 2012, when Arctic sea ice declined to a record minimum. The average net carbon uptake rate of the phytoplankton in polycarbonate (PC) bottles in the closed MP was 3.24 mg C · m^?3 · h^?1 (SD = ±1.12 mg C · m^?3 · h^?1), while that in the open MP was 1.3 mg C · m^?3 · h^?1 (SD = ±0.05 mg C · m^?3 · h^?1). The net production rate of total MAAs in incubated PC bottles was highest (1.44 (SD = ±0.24) ng C · L^?1 · h^?1) in the open MP and lowest (0.05 (SD = ±0.003) ng C · L^?1 · h^?1) in the closed MP. The net production rate of shinorine and palythine in incubated PC bottles at the open MP presented significantly high values 0.76 (SD = ±0.12) ng C · L^?1 · h^?1and 0.53 (SD = ±0.06) ng C · L^?1 · h^?1. Our results showed that high net production rate of MAAs in the open MP was enhanced by a combination of osmotic and UVR stress and that in situ net production rates of individual MAA can be determined using ¹³C tracer in MPs in Arctic sea ice.     

Purification of R‐phycoerythrin from Porphyra haitanensis (Bangiales,Rhodophyta) using expanded‐bed absorption1

R‐phycoerythrin (R‐PE) was purified from leafy gametophyte of Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) by a simple, scaleable procedure. Initially, phycobiliproteins were extracted by repeated freeze‐thaw cycles, resulting in release from the algal cells by osmotic shock. Next, R‐PE was recovered by applying the crude extract with a high concentration of (NH₄)₂SO₄ salt directly to the expanded‐bed columns loaded with phenyl‐sepharose. An expanded‐bed volume twice the settled‐bed volume was maintained; then low (NH₄)₂SO₄ concentration was used to develop the column. After two rounds of hydrophobic interaction chromatography (HIC), R‐PE was purified by anion‐exchange column. The method was also successful with free‐living conchocelis of P. haitanensis. The purified R‐PE was identified with electrophoresis, and absorption and fluorescence emission spectroscopy. The results were in agreement with those previously reported. The yield with a spectroscopic purity (OD₅₆₅/OD₂₈₀) higher than 3.2 (the ratio of A₅₆₅/A₆₂₀ ≤ 0.02) was 1.4 mg · g^?1 of leafy gametophyte of P. haitanensis. For the free‐living conchocelis of P. haitanensis extract, R‐PE could be purified successfully with only one round of HIC. The yield with a spectroscopic purity (OD₅₆₅/OD₂₈₀) higher than 3.2 (the ratio of A₅₆₅/A₆₂₀ ≤ 0.02) was 5.0 mg · g^?1 of free‐living conchocelis of P. haitanensis. The method described here is a scaleable technology that allows a large quantity of R‐PE to be recovered from the unclarified P. haitanensis crude extract. It is also a high protein recovery technology, reducing both processing costs and times, which enhances the value of this endemic Porphyra of China.     

The influence of light on copper‐limited growth of an oceanic diatom,Thalassiosira oceanica (Coscinodiscophyceae)

Thalassiosira oceanica (CCMP 1005) was grown over a range of copper concentrations at saturating and subsaturating irradiance to test the hypothesis that Cu and light were interacting essential resources. Growth was a hyperbolic function of irradiance in Cu‐replete medium (263 fmol Cu′ · L^?1) with maximum rates achieved at 200 μmol photons · m^?2 · s^?1. Lowering the Cu concentration at this irradiance to 30.8 fmol Cu′ · L^?1 decreased cellular Cu quota by 7‐fold and reduced growth rate by 50%. Copper‐deficient cells had significantly slower (P < 0.0001) rates of maximum, relative photosynthetic electron transport (rETR_max) than Cu‐sufficient cells, consistent with the role of Cu in photosynthesis in this diatom. In low‐Cu medium (30.8 fmol Cu′ · L^?1), growth rate was best described as a positive, linear function of irradiance and reached the maximum value measured in Cu‐replete cells when irradiance increased to 400 μmol photons · m^?2 · s^?1. Thus, at high light, low‐Cu concentration was no longer limiting to growth: Cu concentration and light interacted strongly to affect growth rate of T. oceanica (P < 0.0001). Relative ETR_max and Cu quota of cells grown at low Cu also increased at 400 μmol photons · m^?2 · s^?1 to levels measured in Cu‐replete cells. Steady‐state uptake rates of Cu‐deficient and sufficient cells were light‐dependent, suggesting that faster growth of T. oceanica under high light and low Cu was a result of light‐stimulated Cu uptake.     

The CO2‐concentrating mechanism in the bloom‐forming cyanobacterium Microcystis aeruginosa (Cyanophyceae) and effects of UVB radiation on its operation1

The bloom‐forming cyanobacterium Microcystis aeruginosa (Kütz.) Kütz. 854 was cultured with 1.05 W · m^?2 ultraviolet‐B radiation (UVBR) for 3 h every day, and the CO₂‐concentrating mechanism (CCM) within this species as well as effects of UVBR on its operation were investigated. Microcystis aeruginosa 854 possessed at least three inorganic carbon transport systems and could utilize external HCO₃^? and CO₂ for its photosynthesis. The maximum photosynthetic rate was approximately the same, but the apparent affinity for dissolved inorganic carbon was significantly decreased from 74.7 μmol · L^?1 in the control to 34.7 μmol · L^?1 in UVBR‐treated cells. At 150 μmol · L^?1 KHCO₃ and pH 8.0, Na⁺‐dependent HCO₃^? transport contributed 43.4%–40.2% to the photosynthesis in the control and 34.5%–31.9% in UVBR‐treated cells. However, the contribution of Na⁺‐independent HCO₃^? transport increased from 8.7% in the control to 18.3% in UVBR‐treated cells. The contribution of CO₂‐uptake systems showed little difference: 47.9%–51.0% in the control and 49.8%–47.2% in UVBR‐treated cells. Thus, the rate of total inorganic carbon uptake was only marginally affected, although UVBR had a differential effect on various inorganic carbon transporters. However, the number of carboxysomes in UVBR‐treated cells was significantly decreased compared to that in the control.     

15N MEASUREMENTS OF AMMONIUM AND NITRATE UPTAKE BY ULVA FENESTRATA (CHLOROPHYTA) AND GRACILARIA PACIFICA (RHODOPHYTA): COMPARISON OF NET NUTRIENT DISAPPEARANCE,RELEASE OF AMMONIUM AND NITRATE,AND 15N ACCUMULATION IN ALGAL TISSUE 1

Ammonium and nitrate uptake rates in the macroalgae Ulva fenestrata (Postels and Ruprecht) (Chlorophyta) and Gracilaria pacifica (Abbott) (Rhodophyta) were determined by ¹⁵N accumulation in algal tissue and by disappearance of nutrient from the medium in long‐term (4–13 days) incubations. Nitrogen‐rich algae (total nitrogen> 4% dry weight [dw]) were used to detect isotope dilution by release of inorganic unlabeled N from algal thalli. Uptake of NH₄ ⁺ was similar for the two macroalgae, and the highest rates were observed on the first day of incubation (45 μmol N·g dw ^{? 1}·h ^{? 1} in U. fenestrata and 32 μmol N·g dw ^{? 1}·h ^{? 1} in G. pacifica). A significant isotope dilution (from 10 to 7.9 atom % enrichment) occurred in U. fenestrata cultures during the first day, corresponding to a NH₄ ⁺ release rate of 11 μmol N·g dw ^{? 1}·h ^{? 1}. Little isotope dilution occurred in the other algal cultures. Concurrently to net NH₄ ⁺ uptake, we observed a transient free amino acid (FAA) release on the first day in both macroalgal cultures. The uptake rates estimated by NH₄ ⁺ disappearance and ¹⁵N incorporation in algal tissue compare well (82% agreement, defined as the percentage ratio of the lower to the higher rate) at high NH₄ ⁺ concentrations, provided that isotope dilution is taken into account. On average, 96% of added ¹⁵NH₄ ⁺ was recovered from the medium and algal tissue at the end of the incubation. Negligible uptake of NO₃ ^? was observed during the first 2–3 days in both macroalgae. The lag of uptake may have resulted from the need for either some N deprivation (use of NO₃ ^? pools) or physiological/metabolic changes required before the uptake of NO₃ ^? . During the subsequent days, NO₃ ^? uptake rates were similar for the two macroalgae but much lower than NH₄ ⁺ uptake rates (1.97–3.19 μmol N·g dw ^{? 1}·h ^{? 1}). Very little isotope dilution and FAA release were observed. The agreement between rates calculated with the two different methods averaged 91% in U. fenestrata and 95% in G. pacifica. Recovery of added ¹⁵NO₃ ^? was virtually complete (99%). These tracer incubations show that isotope dilution can be significant in NH₄ ⁺ uptake experiments conducted with N‐rich macroalgae and that determination of ¹⁵N atom % enrichment of the dissolved NH₄ ⁺ is recommended to avoid poor isotope recovery and underestimation of uptake rates.     

RAPID AMMONIUM‐ AND NITRATE‐INDUCED PERTURBATIONS TO CHL a FLUORESCENCE IN NITROGEN‐STRESSED DUNALIELLA TERTIOLECTA (CHLOROPHYTA)1

When NH₄ ⁺ or NO₃ ^? was supplied to NO₃ ^? ‐stressed cells of the microalga Dunaliella tertiolecta Butcher, immediate transient changes in chl a fluorescence were observed over several minutes that were not seen in N‐replete cells. These changes were predominantly due to nonphotochemical fluorescence quenching. Fluorescence changes were accompanied by changes in photosynthetic oxygen evolution, indicating interactions between photosynthesis and N assimilation. The magnitude of the fluorescence change showed a Michaelis‐Menten relationship with half‐saturation concentration of 0.5 μM for NO₃ ^? and 10 μM for NH₄ ⁺ . Changes in fluorescence responses were characterized in D. tertiolecta both over 5 days of N starvation and in cells cultured at a range of NO₃ ^? ‐limited growth rates. Variation in responses was more marked in starved than in limited cells. During N starvation, the timing and onset of the fluorescence responses were different for NO₃ ^? versus NH₄ ⁺ and were correlated with changes in maximum N uptake rate during N starvation. In severely N‐starved cells, the major fluorescence response to NO₃ ^? disappeared, whereas the response to NH₄ ⁺ persisted. N‐starved cells previously grown with NH₄ ⁺ alone showed fluorescence responses with NH₄ ⁺ but not NO₃ ^? additions. The distinct responses to NO₃ ^? and NH₄ ⁺ may be due to the differences between regulation of the uptake mechanisms for the two N sources during N starvation. This method offers potential for assessing the importance of NO₃ ^? or NH₄ ⁺ as an N source to phytoplankton populations and as a diagnostic tool for N limitation.     

Unravelling growth trajectories from complicated otoliths – the case of Brazilian codling Urophycis brasiliensis

Uncertainty regarding the age determination of the Brazilian codling Urophycis brasiliensis has hampered its stock assessment. Transverse sections of otoliths displayed up to seven (in males) and 12 (in females) alternate opaque and translucent bands that could not be conclusively validated as annuli, resulting in unrealistically high ages of first maturity (A₅₀) (A_50male = 4·5 years and A_50female = 6 years). Therefore, growth was described by the von Bertalanffy (VB) model using an alternative approach that combined microstructure data (daily growth increments) and a fixed asymptotic total length (L_∞). This approach was supported by applying it to two other co‐occurring species, the whitemouth croaker Micropogonias furnieri and the king weakfish Macrodon atricauda, for which daily and annual ring formation has previously been validated. The sensitivity to realistic errors associated with the choice of the L_∞ and the daily increment readings was shown to be low. The results show that U. brasiliensis has a fast growth rate (K_male = 1·19 year^?1, K_female = 0·71 year^?1) and early maturation (A_50male = 1·1–1·5 years, A_50female = 1·6–1·8 years); typical life‐history traits for a sub‐tropical coastal gadiform. This novel study offers an alternative approach for age and growth reconstruction for species with complex patterns of opaque and translucent bands provided that daily growth increments in the yearlings can be counted and L_∞ reliably estimated.     

Heterotrophic nitrification by <Emphasis Type="Italic">Achromobacter xylosoxidans</Emphasis> S18 isolated from a small-scale slaughterhouse wastewater

Heterotrophic carbon utilizing microbes were acclimatized in the laboratory by inoculating sludge collected from the waste discharge pond of a small-scale rural abattoir in India in a nutrient solution intermittently fed with glucose and ammonium chloride. Cultures of 10 well-developed isolates were selected and grown in a basal medium containing glucose and ammonium chloride. Culture supernatants were periodically analyzed for ammonium nitrogen (NH₄ ⁺-N) and chemical oxygen demand (COD). Polyphasic taxonomic study of the most active nitrifier (S18) was done. Half saturation concentration (K _s), maximum rate of substrate utilization (k), yield coefficient (Y) and decay coefficient (K _d) were determined from the Lineweaver–Burk plot using the modified Monod equation. S18 was able to remove 97 ± 2% of (NH₄ ⁺-N) and 88 ± 3% of COD. Molecular phylogenetic study supported by physiological and biochemical characteristics assigned S18 as Achromobacter xylosoxidans. Nitrification activity of A. xylosoxidans was demonstrated for the first time, while interestingly, the distinctive anaerobic denitrification property was preserved in S18. K _s values were determined as 232.13 ± 1.5 mg/l for COD reduction and 2.131 ± 1.9 mg/l for NH₄ ⁺-N utilization. Yield coefficients obtained were 0.4423 ± 0.1134 mg of MLVSS/mg of COD and 0.2461 ± 0.0793 mg of MLVSS/mg of NH₄ ⁺-N while the decay coefficients were 0.0627 ± 0.0013 per day and 0.0514 ± 0.0008 per day, respectively. After a contact period of 24 h, 650 ± 5 mg/l solids were produced when the initial concentration of COD and NH₄ ⁺-N were 1820 ± 10 mg/l and 120 ± 5.5 mg/l, respectively. This is the first report on the kinetic coefficients for carbon oxidation and nitrification by a single bacterium isolated from slaughterhouse wastewater.     

Luminescent properties of 4‐aminobenzo‐15‐crown‐5 after preferential binding of ferric ions in aqueous solutions

We report a combined approach that introduces the use of 4‐aminobenzo‐15‐crown‐5 (4AB15C5) for the detection of ferric(III) ions by colorimetric, ultraviolet (UV)–visible light absorption, fluorescence, and live‐cell imaging techniques along with density functional theory (DFT) calculations. We have found that 4AB15C5 is sensitive and selective for binding ferric(III) ions in aqueous solutions. DFT calculations using the polarizable continuum model have been used to explain the strong binding of the ferric ion by 4AB15C5 in aqueous solutions. The detection limit in the fluorescence quenching measurements was found to be as low as 50 μM for the ferric ion with a determined Stern–Volmer constant of 1.52 × 10⁴ M^?1. Fluorescence intensity did not change for other ions tested, Fe²⁺, Co²⁺, Mn²⁺, Mg²⁺, Zn²⁺, Ca²⁺, NH₄⁺, Na⁺, and K⁺ ions. Live‐cell fluorescence imaging was also used to check the intracellular variations in ferric ion levels. Our spectroscopic data indicated that 4AB15C5 can bind ferric ions selectively in aqueous solutions.     

Effect of trace elements and optimization of their composition for the nitrification of a heterotrophic nitrifying bacterium, <Emphasis Type="Italic">Acinetobacter harbinensis</Emphasis> HITLi7<Superscript>T</Superscript>, at low temperature

The effects of trace elements on ammonium degradation performance and extracellular polymeric substances (EPS) secretion of Acinetobacter harbinensis HITLi7^T at low temperature were investigated. Response surface methodology (RSM) was applied to obtain the optimal composition of trace elements and analyze their correlation. In this study, the results indicated that the ammonium removal performance could be enhanced by the presence of 0.1 mg L^?1 Fe, Mn, or B in pure cultivation. When the concentrations of Fe and Mn were 0.2 mg L^?1, the ammonium removal rates of the novel strain HITLi7^T were 0.49 ± 0.01 mg L^?1·h^?1 and 0.58 ± 0.01 mg L^?1·h^?1, respectively, while it was the low concentration of 0.05 mg L^?1 B that showed the maximum ammonium removal rate (0.56 ± 0.02 mg L^?1·h^?1) of strain HITLi7^T. The regression model was obtained and the optimal formulation of trace elements was: B 0.064 mg L^?1, Fe 0.12 mg L^?1, and Mn 0.1 mg L^?1. Based on these values, the experimental ammonium removal rate could reach 0.59 mg L^?1·h^?1, which matched well with the predicted response. The study also found that the addition of trace elements, causing high ammonium removal rates, resulted in a high polysaccharide (PS) ratio in the EPS secreted by Acinetobacter harbinensis HITLi7^T. Especially under the optimal conditions, the PS ratio reached the highest value of 49.9%.     

CULTIVATION OF ARTHROSPIRA (SPIRULINA) PLATENSIS (CYANOPHYCEAE) BY FED‐BATCH ADDITION OF AMMONIUM CHLORIDE AT EXPONENTIALLY INCREASING FEEDING RATES1

Arthrospira (Spirulina) platensis (Nordstedt) Gomont was cultivated under light‐limited conditions in 5‐L open tanks by daily supplying NH₄Cl as nitrogen source. Exponentially increasing feeding rates were adopted to prevent ammonia toxicity. The total feeding time (T) was varied between 12 and 20 days, and the starting (m₀) and total (m_T) quantities of the nitrogen source per unit reactor volume were varied in the ranges 0.19–1.7 mM and 2.3–23.1 mM, respectively. This intermittent addition of the nitrogen source prevented ammonia from reaching inhibitory levels and ensured final cell concentrations (X_m) and cell productivities (P_x) comparable with those of batch runs with KNO₃. Moreover, the lower nitrogen addition due to the use of NH₄Cl rather than KNO₃ allowed for higher nitrogen‐to‐cell conversions (Y_x/n). These results were evaluated using three‐factor, five‐level, central composite experimental planning, combined with the response surface methodology, selecting T, m₀, and m_T as the independent variables and X_m, P_x, and Y_x/n as the response variables. This approach allowed us to identify, through the simultaneous optimization of the variables, T=16 days, m₀=1.7 mM, and m_T=21.5 mM as the best conditions for A. platensis cultivation at 72 μmol photons·m^?2·s^?1. Under these conditions, a maximum cell concentration of 1239 mg ·L^?1 was obtained, which is a value comparable with that obtained using KNO₃ as nitrogen source and nearly coincident with the theoretical one estimated by the response surface methodology.     

Different fermentation strategies by Schizochytrium mangrovei strain pq6 to produce feedstock for exploitation of squalene and omega‐3 fatty acids

Schizochytrium mangrovei strain PQ 6 was investigated for coproduction of docosahexaenoic acid (C22: 6ω‐3, DHA ) and squalene using a 30‐L bioreactor with a working volume of 15 L under various batch and fed‐batch fermentation process regimes. The fed‐batch process was a more efficient cultivation strategy for achieving higher biomass production rich in DHA and squalene. The final biomass, total lipid, unsaponifiable lipid content, and DHA productivity were 105.25 g · L^?1, 43.40% of dry cell weight, 8.58% total lipid, and 61.66 mg · g^?1 · L^?1, respectively, after a 96 h fed‐batch fermentation. The squalene content was highest at 48 h after feeding glucose (98.07 mg · g^?1 of lipid). Differences in lipid accumulation during fermentation were correlated with changes in ultrastructure using transmission electron microscopy and Nile Red staining of cells. The results may be of relevance to industrial‐scale coproduction of DHA and squalene in heterotrophic marine microalgae such as Schizochytrium .     

NH4 : NO3 nutrition influence on biomass productivity and root respiration of poplar and willow clones

Nitrogen fertilization often improves the yield of intensively managed, short‐rotation coppices. However, information of N nutrition form on the growth of common species and clones used for biomass production is limited. Thus, this study aims at evaluating N form effects on the growth of two Salicaceae clones. Cuttings of the poplar clone Max 4 (Populus maximovizcii × P. nigra) and the willow clone Inger (Salix triandra × S. viminialis) were fertilized in a pot experiment with four ratios of nitrate (NO₃^?) to ammonium (50%, 62.5%, 75% and 87.5% NO₃^? balanced with ammonium (NH₄⁺) to constant total N) for one growing season and under stable soil pH. Plants were harvested for analysis of biomass and morphology of leaves, stem and roots. Respiration of fine and coarse roots (RR) was determined and related to biomass growth. Salix cv. Inger accumulated more total dry matter than Populus cv. Max 4. In both Salicaceae clones, the total biomass was significantly influenced by the nitrate ratio and greatest in plants fertilized with 50% NO₃^? of the total N supply. Both clones possess a different leaf and root morphology, but no significant influence of the NO₃^? ratio on the morphology was found. Fine RR rates differed significantly between clones, with significantly greater fine RR in Max 4; 87.5% NO₃^? fertilization increased the fine RR. Fine RR and total accumulated plant biomass were closely related. Our study is the first to show the tremendous influence of fine root respiration, especially including the carbon‐intensive reduction of NO₃^? to NH₄⁺, on the aboveground growth of Salicaceae clones. Ways to improve yield in SRC are thus to lower the assimilate consumption by fine roots and to match fertilization regimes to the used clones or vice versa.