Nitrate toxicity toPenaeus monodon protozoea

Increased levels of nitrate occur in natural waters due to pollution, and in aquaculture systems from nitrification and addition of microalgal cultures for feeding. Static bioassays showed that significant mortality of larvalPenaeus monodon (Fabricius) occurred within 40 h at nitrate concentrations as low as 1 mg NO ₃ ^- l^–1. Sublethal effects of this concentration resulted in changes to ganglionic neuropiles and muscles. At higher concentrations (10 and 100 mg NO ₃ ^- l^–1), additional tissues were affected including the hypodermis, midgut and proventriculus. This is the first report of toxicity to a marine organism of nitrate at concentrations normally present in enclosed seawater and mariculture systems. The results are discussed in terms of management of culture systems and of natural marine ecosystems containing elevated levels of nitrate.     

Inhibition of diatom photosynthesis by germanic acid: Separation of diatom productivity from total marine primary productivity

Germanic acid was shown to inhibit photosynthetic ¹⁴CO₂ uptake in marine diatoms. Inhibition was not complete even at concentrations of 20 mg Ge/1 nor in cultures incubated for extended periods of time (up to 24 h). The decrease in photosynthesis due to Ge(OH)₄ was independent of the stage of growth of the diatom culture. At 0.5 and 1.0 mg Ge/1, the degree of inhibition was dependent on the concentration of Si(OH)₄ in the medium. At 5 and 10 mg Ge/1, inhibition was not affected by Si(OH)₄ concentrations as high as those found in the sea-120 g-at Si/1. The effect of Ge(OH)₄ on photosynthesis is specific for diatoms; other marine phytoplankton were not inhibited. In mixed cultures of diatoms and marine flagellates, the reduction in ¹⁴CO₂ fixation upon addition of Ge(OH)₄ was used to calculated the proportion of diatom photosynthesis to total photosynthesis, and calculated proportions agreed well with actual proportions. Inhibition by Ge(OH)₄ was also used to estimate the percent of diatom photosynthesis in a natural marine community, and this was compared with the diatom portion of the crop. Diatom photosynthesis was higher than one might expect from crop figures, although both diatom photosynthesis and diatom numbers in the crop were low.     

Dark CO2 uptake by the diatom Chaetoceros simplex in response to nitrogen pulsing

In a continuing investigation of dark CO₂ uptake by nitrogen-limited cultures of the marine diatom Chaetoceros simplex (Bbsm), we expanded on several of our earlier conclusions regarding the potential application of this physiological response for measuring the degree and type of nitrogen limitation in phytoplankton populations. First, the duration over which the maximal enhancement of dark ¹⁴CO₂ uptake was sustained after NH ₄ ⁺ enrichment was a function both of the concentration of added NH ₄ ⁺ and the standing crop of phytoplankton nitrogen — in effect, the total N demand. Second, pulsing with NH ₄ ⁺ for a given degree of N-limitation always produced the same level of enhanced dark CO₂ uptake regardless of whether the cultures were preconditioned with oxidized or reduced nitrogen. In contrast, urea pulsing led to reduced dark CO₂ uptake, but the effect was most pronounced in cells grown on NO ₃ ^– . And third, the assay could be used to distinguish readily between no, moderate, and severe N limitation. The degree of severe N limitation was quantitatively correlated with the degree of enhanced dark CO₂ uptake, but this relationship was not so clear in the region of moderate N limitation. The main advantage of the assay is that it is a relatively simple and effective alternative to more complicated techniques for gauging the degree and form of N limitation in phytoplankton. Further evaluation will be required, both in the laboratory and field, before the assay can be calibrated for quantitative use.Contribution No. 5982 from the Woods Hole Oceanographic Institution     

Acid-base responses to lethal aquatic hypercapnia in three marine fishes

Ocean sequestration of CO₂ is proposed as a possible measure to mitigate environmental changes due to the increasing atmospheric concentration of the gas. However, toxic effects of CO₂ on marine organisms are poorly understood. We therefore studied acid–base responses and mortality during exposure to fatal levels of CO₂ in three marine fishes (Japanese flounder, Paralichthys olivaceus; yellowtail, Seriola quinqueradiata; and starspotted dogfish, Mustelus manazo). The teleosts died during exposure to seawater equilibrated with a gas mixture containing 5% CO₂ (water PCO₂ 4.95 kPa); 100% mortality occurred within 8 h for yellowtail and within 48 h for flounder. Only 20% mortality was recorded at 72 h for the dogfish during exposure to 7% CO₂ (water PCO₂ 6.96 kPa). Arterial pH (pH_a) initially decreased, but completely recovered within 1–24 h for the teleosts at 1% and 3%, although the recovery was slower and complete only at 1% (water PCO₂ 0.99 kPa) for the dogfish. During exposure to 5%, the flounder died after the pH_a had been completely restored, suggesting that the mortality was not due to plasma acidosis. During exposure to 1% hypercapnia, plasma [Cl^–] appeared to be the main counter ion to balance increases in plasma [HCO₃^-]. There was a 1:1 stoichiometry for the rise in [HCO₃^-] and the fall in [Cl^–] for the teleosts, whereas the ratio was 1:0.4 for the dogfish at 1% CO₂. At the higher levels of hypercapnia, the rise in [HCO₃^-] consistently exceeded the fall in [Cl^–], and plasma [Na⁺] significantly increased.These results do not agree with the generally accepted model for acid–base regulation in marine fish in which Na⁺/H⁺ exchangers are assumed to play a predominant role, and indicate that an acid–base regulatory mechanism differs between teleost and elasmobranch fishes, as well as the intensity of acidic stress.Communicated by T. Ikeda, Hakodate     

Iron-mediated effects on nitrate reductase in marine phytoplankton

The potential activity of nitrate reductase was determined in uni-algal cultures in the laboratory and in natural marine phytoplankton assemblages. In the laboratory bioassays, distinct differences in nitrate reductase activity were observed in iron replete versus depleted cultures for Emiliania huxleyi, Isochrysis galbana and Tetraselmis sp. Cells from iron-depleted cultures had 15 to 50 percent lower enzyme activity than those from iron-replete cultures. Upon addition of iron, nitrate reductase activity was enhanced in depleted cells up to levels comparable to those of the replete cells. Bioassays in the northern North Sea conducted in 1993, under low iron conditions, demonstrated similar results. Upon addition of 2.5 nM iron, a distinct enhancement, to a maximum of three times, of nitrate reductase activity was observed within 32 h after addition. Therefore, iron can stimulate nitrate reductase activity. In spite of the clean techniques used, some nitrate reductase activity was always observed. Iron deficiency was shown to impair nitrate reductase activity, but it is unlikely that nitrate reduction would cease completely.     

Selective reduction of NO by photo-SCR with ammonia in an annular fixed-film photoreactor

Gaseous NO was photocatalytically reduced at room temperature by photo-assisted selective catalytic reduction (photo-SCR) with ammonia over TiO₂ in this study. NO reduction efficiency and N₂ selectivity were determined from gases composition at the outlet stream of photoreactor. Effect of operating conditions, e.g. light intensity and inlet concentrations of ammonia and oxygen, on the NO reduction efficiency and N₂ selectivity were discussed to determine the feasible operating condition for photocatalytic reduction of NO. Experimental results showed that selective catalytic reduction of NO with ammonia over TiO₂ in the presence of oxygen was a spontaneous reaction in dark. The photoirradiation on the TiO₂ surface caused remarkable photocatalytic reduction of NO to form N₂, NO₂, and N₂O under 254 nm UV illuminations, while almost 90% of N₂ selectivity was achieved in this study. The ammonia and oxygen molecules played the roles of reductant and oxidant for NO reduction and active sites regeneration, respectively. The reduction of NO was found to be increased with the increase of inlet ammonia and oxygen concentrations until specific concentrations because of the limited active sites on the surface of TiO₂. The kinetic model proposed in this study can be used to reasonably describe the reaction mechanism of photo-SCR.     

Physiological responses of the calcifying rhodophyte, <Emphasis Type="Italic">Corallina officinalis</Emphasis> (L.), to future CO<Subscript>2</Subscript> levels

Future atmospheric CO₂ levels will most likely have complex consequences for marine organisms, particulary photosynthetic calcifying organisms. Corallina officinalis L. is an erect calcifying macroalga found in the inter- and subtidal regions of temperate rocky coastlines and provides important substrate and refugia for marine meiofauna. The main goal of the current study was to determine the physiological responses of C. officinalis to increased CO₂ concentrations expected to occur within the next century and beyond. Our results show that growth and production of inorganic material decreased under high CO₂ levels, while carbonic anhydrase activity was stimulated and negatively correlated to algal inorganic content. Photosynthetic efficiency based on oxygen evolution was also negatively affected by increased CO₂. The results of this study indicate that C. officinalis may become less competitive under future CO₂ levels, which could result in structural changes in future temperate intertidal communities.     

Inorganic nitrogen metabolism in marine bacteria: Nitrate uptake and reduction in a marine pseudomonad

Growth experiments in batch cultures indicated that the uptake of nitrate by the marine pseudomonad PL1 was inhibited in the presence of ammonia provided that the ammonia concentration was higher than 1 mM. At ammonia concentrations of less than about 1 mM, however, both nitrate and ammonia were utilised simultaneously. The saturation constants for nitrate and ammonia uptake were both 2.6x10^-4 M, and similar to the Michaelis constants of nitrate reductase for nitrate (2.9x10^-4 M) and glutamine synthetase for ammonia (2x10^-4 M). Nitrate reductase activity linked to NADH was detected in chemostat-grown cultures with nitrate as nitrogen source, and in cultures containing limiting concentrations of nitrate and ammonia, ammonia or glutamate. Enzyme synthesis appeared to be repressed in cultures containing an excess of ammonia or glutamate. Chemostat cultures utilised ammonia or glutamate in preference to nitrate, while there was no marked preference between ammonia and glutamate.     

High CO<Subscript>2</Subscript> adsorption by amino-modified bio-spherical cellulose nanofibres aerogels

Climate change has become increasingly serious due to the greenhouse effect. It is therefore necessary to control the content of greenhouse gases such as carbon dioxide in the atmosphere, using, for instance, CO₂-adsorbing materials. Here, we synthesized ultra-lightweight and spherical cellulose nanofibres aerogels by a suspension titration method using an efficient amination process. These functional materials with high porosity, higher than 96.54%, and three-dimensional network structure, were prepared by freeze-drying spherical cellulose nanofibres hydrogel. Their maximum CO₂ adsorption capacity reaches 1.78 mmol/g, and they show excellent regeneration, of more than 10 cycles. This synthesis of bioaerogels represents a new method for the preparation of bio-CO₂ adsorbents.     

Ionic liquids for the inhibition of gas hydrates. A review

The formation of gas hydrates is a major issue during the operation of oil and gas pipelines, because gas hydrates cause plugging, thereby disrupting the normal oil and gas flows. A solution is to inject gas hydrate inhibitors such as ionic liquids. Contrary to classical inhibitors, ionic liquids act both as thermodynamic inhibitors and hydrate inhibitors, and as anti-agglomerates. Imidazolium-based ionic liquids have been found efficient for the inhibition of CO₂ and CH₄ hydrates. For CO₂ gas hydrates, N-ethyl-N-methylmorpholinium bromide showed an average depression temperature of 1.72 K at 10 wt% concentration. The induction time of 1-ethyl-3-methyl imidazolium bromide is 36.3 h for CO₂ hydrates at 1 wt% concentration. For CH₄ hydrates, 1-ethyl-3-methyl-imidazolium chloride showed average depression temperature of 4.80 K at 40 wt%. For mixed gas hydrates of CO₂ and CH₄, only quaternary ammonium salts have been studied. Tetramethyl ammonium hydroxide shifted the hydrate liquid vapour equilibrium to 1.56 K at 10 wt%, while tetrabutylammonium hydroxide showed an induction time of 0.74 h at 1 wt% concentration.

     

Photosynthesis and carbon storage between tides in a brown alga, Fucus vesiculosus

Intertidal macroalgae may spend a significant part of their lives in air. During photosynthesis in air, they encounter much lower concentrations of inorganic carbon than in seawater. Because they accumulate inorganic carbon from seawater, we investigated whether they similarly accumulate it from air. We measured photosynthesis in the intertidal species Fucus vesiculosus L. during 1990 and 1991 with a gas-phase O₂ electrode or CO₂-exchange apparatus in air and with a liquid-phase O₂ electrode in seawater. Maximum rates were rapid and similar in air and seawater regardless of the method. Tissue from seawater could carry on photosynthesis in CO₂-free air, indicating that carbon was stored in the tissue. After 2 h, this store was depleted and photosynthesis ceased. Supplying CO₂ in air replenished the store. Under identical conditions, terrestrial C₃ and C₄ species showed no evidence of this store, but a CAM (crassulacean acid metabolism) species did. However, in contrast to the CAM behavior, F. vesiculosus did not store CO₂ significantly in the dark. We found a small acid-releasable pool of carbon in the tissue that disappeared as photosynthesis depleted the carbon store. However, the pool was too small to account for the total carbon stored. While CO₂ was being acquired or released from the store in the light, photosynthesis was not inhibited by 21% O₂. These results indicate that there are two parallel paths for the supply of CO₂ to photosynthesis. The first depends on inorganic carbon in seawater or in air and supports rapid photosynthesis. The second involves CO₂ slowly released from an organic intermediate. The release protects CO₂ fixation from the inhibitory effects of 21% O₂. Photosynthesis in F. vesiculosus thus appears to be C₃-like in its rapid fixation of CO₂ from a small inorganic pool into phosphoglycerate. However, it is C₄-like in its pre-fixation of carbon in an organic pool in the light, and is CAM-like in its ability to slowly use this pool as a sole source of CO₂. The organic pool may serve to protect photosynthetic CO₂ fixation against the inhibitory effects of O₂ in air and in the boundary layer in seawater. Received: 6 March 1998 / Accepted: 16 October 1998     

RQ of benthic marine invertebrates

This study investigated an incubation method which employed simultaneous measurement of CO₂ production and O₂ consumption rates to calculate the RQ (respiratory quotient; CO₂ production rate: O₂ consumption rate) of individual benthic marine invertebrates. Carbon dioxide production rates were calculated from changes in CO₂ concentration determined using seawater pH. O₂ consumption rates were calculated from changes in O₂ concentration with a correction applied for O₂ flux across the air/water interface due to gaseous exchange. Species examined were Triphyllozoon sp. cf. moniliferum (MacGillivray 1860), a bryozoan; Herdmania momus (Savigny), a solitary ascidian; Poneroplax albida (Blainville 1825), a chiton; and Haliotis roei (Gray 1826), an abalone. Six individuals of each were collected on 14 November 1985 from the limestone walls of a cave in a nearshore reef off Marmion, Western Australia. After acclimation for 6 h in experimental conditions, rates of CO₂ production and O₂ consumption were measured. A minimum period of 4 h was required to obtain consistent RQ values for each species. The standard error (SE) of the (calculated) RQ ratio was 14 to 33% of the mean in incubations of 4 h, and less than 14% in incubations of 4 to 12 h. The RQ is commonly used as an indicator of unknown catabolic substrates by comparing it with biochemically determined limits for known substrates. This study provides a strong argument against using the RQ of individual animals to draw any conclusions about catabolic substrates. Unexplained variation in the components of the RQ of an individual, measured over short time periods, and the potential involvement of stored reserves in catabolism, over longer time periods, obscure the relationship between the RQ of individual animals and the ratio's biochemically determined limits.     

New denitrifying bacteria isolated from Red Sea sediments

Sediments of various newly discovered deeps in the Red Sea were analyzed for the occurrence of denitrifying bacteria. The samples were collected in 1972 during the Valdivia cruises. Among the 27 different samples investigated, 17 revealed both coccoid and rod-shaped bacteria when enriched in complex nutrient broth (with 10% NaCl). Denitrifiers were recorded abundantly in the sediments, their population decreasing from some 10⁶/g in the surface material to only a few in the subsediment. A total of 16 pure cultures of denitrifying bacteria were isolated from the Suakin-and Thetis-deeps and studied morphologically, physiologically and biochemically. Genetics (molar percentage of guanine plus cytosine, % GC) and numerical taxonomy were included to reveal relationships and improve taxonomic classification. Fifteen isolates were described as Gram-negative, aerobic and facultative anaerobic (with NO ₃ ^- as H⁺-acceptor), polarly flagellated rods (Pseudomonas spp.); one was an inmotile, Gram-positive, facultative anaerobic coccus. None of the 15 Pseudomonas-isolates could be identified with one of the denitrifying species so far described and recognized. The strains should be regarded as hitherto undescribed denitrifying marine bacteria.Supported by the Bundesministerium für Forschung und Technologie, Bonn, Germany (FRG).     

Degradation of ammonium dinitramide (ADN) in digested sewage sludge under strict anaerobic conditions

The biodegradation of one popular nitramine energetics, ammonium dinitramide (ADN) by mixture of denitrifying bacterial species was investigated. ADN was observed to be effectively mineralized in the anaerobic mixed culture. The initial ADN concentration of 250 mg/L was reduced to non‐detectable levels (> 99% removal efficiency) in 5 days of incubation under anaerobic conditions. Final products generated from anaerobic degradation of nitramine energetics by anaerobic metabolism were NH₄ ⁺, CH₄, and CO₂ that were released to the environment with the denitrifiers’ growth. In addition, it was found that the activity of denitrifiers was inhibited by high concentration of ammonia generated through the degradation reactions of energetic nitrites.     

Effect of biochar addition on short-term N<Subscript>2</Subscript>O and CO<Subscript>2</Subscript> emissions during repeated drying and wetting of an anthropogenic alluvial soil

Agricultural soils are an important source of greenhouse gases (GHG). Biochar application to such soils has the potential of mitigating global anthropogenic GHG emissions. Under irrigation, the topsoils in arid regions experience repeated drying and wetting during the crop growing season. Biochar incorporation into these soils would change the soil microbial environment and hence affect GHG emissions. Little information, however, is available regarding the effect of biochar addition on carbon dioxide (CO₂) and nitrous oxide (N₂O) emissions from agricultural soils undergoing repeated drying and wetting. Here, we report the results of a 49-day aerobic incubation experiment, incorporating biochar into an anthropogenic alluvial soil in an arid region of Xinjiang Province, China, and measuring CO₂ and N₂O emissions. Under both drying–wetting and constantly moist conditions, biochar amendment significantly increased cumulative CO₂ emission. At the same time, there was a significant reduction (up to ~20 %) in cumulative N₂O emission, indicating that the addition of biochar to irrigated agricultural soils may effectively slow down global warming in arid regions of China.     

Responses of marine benthic microalgae to elevated CO2

Increasing anthropogenic CO₂ emissions to the atmosphere are causing a rise in pCO₂ concentrations in the ocean surface and lowering pH. To predict the effects of these changes, we need to improve our understanding of the responses of marine primary producers since these drive biogeochemical cycles and profoundly affect the structure and function of benthic habitats. The effects of increasing CO₂ levels on the colonisation of artificial substrata by microalgal assemblages (periphyton) were examined across a CO₂ gradient off the volcanic island of Vulcano (NE Sicily). We show that periphyton communities altered significantly as CO₂ concentrations increased. CO₂ enrichment caused significant increases in chlorophyll a concentrations and in diatom abundance although we did not detect any changes in cyanobacteria. SEM analysis revealed major shifts in diatom assemblage composition as CO₂ levels increased. The responses of benthic microalgae to rising anthropogenic CO₂ emissions are likely to have significant ecological ramifications for coastal systems.     

Photosynthetic carbon acquisition in the red alga Gracilaria conferta

In this continuing study on photosynthesis of the marine red alga Gracilaria conferta, it was found that ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in crude extracts had a K _m (CO₂) of 85 M. Since seawater contains only ca. 10 M CO₂, it appears that this alga must possess a CO₂ concetrating system in order to supply sufficient CO₂ to the vicinity of the enzyme. Because this species is a C₃ plant (and thus lacks the C₄ system for concentrating CO₂), but can utilize HCO₃ ^- as an exogenous carbon source, we examined whether HCO₃ ^- uptake could be the initial step of such a CO₂ concetrating system. The surface pH of G. conferta thalli was 9.4 during photosynthesis. At this pH, estimated maximal uncatalyzed HCO₃ ^- dehydration (CO₂ formation) within the unstirred layer was too slow to account for measured phostosynthetic rates, even in the presence of an external carbonic anhydrase inhibitor. This observation, and the marked pH increase in the unstirred layer following the onset of light, suggests that a HCO₃ ^- transport system (probably coupled to transmembrane H⁺/OH^- fluxes) operates at the plasmalemma level. The involvement of surface-bound carbonic anhydrase in such a system remains, however, obscure. The apparent need of marine macroalgae such as G. conferta for CO₂ concentrating mechanisms is discussed with regard to their low affinity of Rubisco to CO₂ and the low rate of CO₂ supply in water. The close similarity between rates of Rubisco carboxylation and measured photosynthesis further suggests that the carboxylase activity, rather than inorganic carbon transport and intercoversion events, could be an internal limiting factor for photosynthetic rates of G. conferta.     

Effect of carbon dioxide concentration on calcification in the red coralline alga Bossiella orbigniana

The relationship between various experimental concentrations of CO₂ and calcification in Bossiella orbigniana (Decaisne) was studied by measuring Ca-45 incorporation into the crystalline matrix. Air containing CO₂ at partial pressures (P_{CO 2}) of 0.04 to 5.5% was bubbled through synthetic seawater in incubation vessels. The resultant pH values in the presence of plants ranged from 6.5 to 8.7. The maximum calcification rate appears to lie between 0.11 and 1.05% P_{CO 2}. The data suggest that calcification is controlled by a biological process that may be sensitive to pH and/or to the relative bicarbonate concentration. The data also suggest that a severalfold increase in CO₂ over the present atmospheric level might lead to increased calcification in this marine alga.