A new type of metabolism chamber for the determination of active and postprandial metabolism offish, and consideration of results for coregonid and salmon juveniles

The optomotor reaction of juvenile Coregonus schinzipalea Val. et Cuv. and Salmo salar L. was utilized to develop a circular tube metabolism chamber to measure oxygen consumption and ammonia excretion as a function of swimming speed. The metabolism chamber with a constant water flow assured the maintenance of stable conditions. The unidirectional movement of fish was measured in a circular tube with a single narrowing. The relationships between the swimming speed and oxygen consumption or ammonia excretion described by exponential equations allowed the extrapolation towards the standard metabolism, i.e., zero swimming speed. For a juvenile coregonid (0.1–0.15 g individual weight, 2.6–2.8 cm total length) standard metabolism at 14° C was estimated as 0.65 mg0₂ g⁻¹ h⁻¹ and 17.3 μg N(NH₃)g⁻¹ h⁻¹, whereas for juvenile salmon (136mg individual weight) respective values at 22° C were 0.047mg0₂g⁻¹h⁻¹ and 0.61 μg N(NH₃)g⁻¹ h⁻¹. The feeding test with juvenile salmon was also performed in this circular chamber, and in both energy and nitrogen budgets after a meal the partitioning could be precisely attributed to standard metabolism, active metabolism and specific dynamic action (in the case of oxygen consumption) or postprandial nitrogen increase.
The new metabolism chamber allowed the relationship between metabolism and swimming velocity of juvenile fish with developed rheotactic response. It could be used with adult fish for similar purposes.     

Impact of gaseous nitrogen deposition on plant functioning

Dry deposition of NH₃ and NO_x (NO and NO₂) can affect plant metabolism at the cellular and whole-plant level. Gaseous pollutants enter the plant mainly through the stomata, and once in the apoplast NH₃ dissolves to form NH₄⁺, whereas NO₂ dissolves to form NO₃⁻ and NO₂⁻. The latter compound can also be formed after exposure to NO. There is evidence that NH₃-N and NO_x-N can be reversibly stored in the apoplast. Temporary storage might affect processes such as absorption rate, assimilation and re-emission. Once formed, NO₃⁻ and NO₂⁻ can be reduced, and NH₄⁺ can be assimilated via the normal enzymatic pathways, nitrate reductase (NR), nitrite reductase and the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle. Fumigation with low concentrations of atmospheric NH₃ increases in vitro glutamine synthetase activity, but whether this involves both or only one of the GS isoforms is still an open question. There seems to be no correlation between fumigation with low concentrations of NH₃ and in vitro GDH activity. The contribution of atmospheric NH₃ and NO₂ deposition to the N budget of the whole plant has been calculated for various atmospheric pollutant concentrations and relative growth rates ( RGRs ). It is concluded that at current ambient atmospheric N concentrations the direct impact of gaseous N uptake by foliage on plant growth is generally small.     

Ecology of Peridinium willei and P. volzii (Dinophyceae) in Danish lakes

The distribution of Peridinium willei and P. volzii was studied in Danish lakes. Both species were confined to lakes with concentrations of Total P < 0.15 mg 1^-1, with the majority of occurrences at Total P concentration between 0.020–0.040 mg 1^-1 and concentrations of PO₄ P between detection limit and 0.040 mg 1^-1. The occurrence of the species in relation to inorganic N compounds (NH₄ N and NO₂+ NO₃ N) was significantly broader for P. willei than for P. volzii: P. willei had an almost even distribution within a wide range of NH₄ N, whereas P. volzii mainly occurred between 0.001 and 0.10 NH₄ N 1^-1. P. willei had an almost even distribution at values beween 0.005 and 0.42 mg NO₂+ NO₃ N 1^-1, whereas P. volzii mainly occurred below 0.050 mg NO₂+ NO₃ N 1¹. P. willei was found at pH values between 4.2 and 8.5, whereas P. volzii was confined to lakes with a slightly basic pH. The study confirmed the broad limits of P. willei and the much more narrow limits of P. volzii in relation to seasonal occurrence and pH, as well as an affinity of the former to ponds and lakes with a rich bottom vegetation. The study also showed, however, that the species were not as widespread and common in recent Danish lake phytoplankton as generally stated by previous authors. The use of different ecological factors to give weight to species separation is discussed. The inclusion of P. volzii in P. willei proposed by Popovsky & Phiester is not supported by the present study, as the two taxa appear to have different ecological tolerances.     

Nitrogen concentration, ammonium/nitrate ratio and NaCl interaction in vegetative and reproductive growth of peanuts

Peanuts ( Arachis hypogaea L. cv. Shulamit) grown with NO₃⁻ and saline water in hydroponics responded positively to addition of nitrogen (N) in their vegetative growth, but not in desert dune sand. In order to clarify these conflicting results, peanut plants were grown in a greenhouse pot experiment with fine calcareous sand. The nutrient solution contained 0 or 50 m M NaCl and 2 or 6 m M N in the form of Ca(NO₃)₂, NH₄NO₃ or (NH₄)₂SO₄. Three replicates were harvested after 48 days (beginning of reproductive stage) and three after 109 days (pod filling). In addition, gynophores were treated with 0, 50, 100, 150 or 200 m M NaCl outside the growth pot to check their sensitivity to salt. Shoot dry weight became greater with increasing NH₄⁺/NO₃⁻ ratio. Increasing the N concentration from 2 to 6 m M did not change shoot dry weight of the NH₄NO₃ or NH₄⁺-fed plants, but caused a reduction in shoot dry weight of NO₃⁻-fed plants. Shoot dry weight was not affected by increasing the NaCl concentration to 50 m M . Salt caused an increase in the number of gynophores per plant and a reduction of the mean pod weight. A NaCl concentration of 100 m M and above reduced gynophore vitality. It is concluded that the salt sensitivity of peanut plants resides mainly in the sensitivity of the reproductive organs.     

The effect of dissolved oxygen and salinity on the toxicity of ammonia to smolts of salmon, Salmo salar L.

The survival of Atlantic salmon smolts on exposure to constant concentrations of ammonia has been measured under laboratory conditions. At concentrations of dissolved oxygen close to the air-saturation value, the 24-h LC50 of un-ionised ammonia is 0.15 mg NH₃1⁻¹ in fresh water (hardness 264 mg 1⁻¹ as CaCO₃) and 0.3 mg NH₃1⁻¹ in 30% sea water; at concentrations of dissolved oxygen of 3.5 mg 1⁻¹ in fresh water and 3.1 mg 1⁻¹ in 30% sea water, the 24-h LC50 is 0.09 mg NH₃ 1⁻¹ and 0.12 mg NH₃ 1⁻¹ respectively; for fish acclimated for 1 day to a concentration of ammonia close to the 24-h median for un-acclimated fish, the median is increased between 38 and 79%, depending on test conditions.     

Methane-dependent nitrate production by a microbial consortium enriched from a cultivated humisol

Abstract A consortium was enriched from a humisol incubated with 3.6 kPa CH₄ and NH₄⁺. This consortium oxidized NH₄⁺ to NO₂⁻ and NO₃⁻ (NO₃⁻/NO₂⁻ ratio about 20) with smaller amounts of N₂O. This oxidation stopped in the stationary phase after depletion of CH₄. CH₃OH or CO₂ did not support oxidation. Growth and resting cell experiments suggested that nitrification was associated with methanotrophic activity and that chemoautotrophic nitrifiers were absent.     

A reaction diffusion model of C-N-S-O species in some arctic sediments

Abstract A reaction diffusion model was used to simulate the mineralization processes in an Arctic sediment. The simulation and the actual sediment were compared in relation to profiles of O₂, NO₃ and NH₄⁺. The site of particulate organic matter (POM) degradation was the single most important factor in fitting the simulation profiles to those of the sediment. It was deduced that most POM degradation occurred close to the sediment surface. When a reasonably good simulation had been obtained, the sensitivity of the model to changes in other parameters was investigated. Increases in POM degradation in the upper sediment resulted in increases in concentration of NH₄⁺ and NO₃⁻, but further increases in POM degradation created anoxic conditions below 3 mm, resulting in decreases in NO₃⁻ concentrations. The model was relatively intensive to changes in POM degradation in the lower sediment layers; increases led to more anoxic conditions and to less NO₃⁻. Increases in the C/N ratio of the POM in the lower sediment layers had little effect; increases in C/N in the upper layers led to a decrease in NH₄⁺ and NO₃⁻. The model was sensitive to changes in the first order rate constant for nitrification, but not for denitrification. Decreases in the K _m for O₂ of the nitrifying bacteria had no effect on the profiles.     

Response of nitrogen metabolism to boron toxicity in tomato plants

Boron (B) toxicity has become important in areas close to the Mediterranean Sea where intensive agriculture has been developed. The objective of this research was to study the effects of B toxicity (0.5 m m and 2.0 m m B) on nitrogen (N) assimilation of two tomato cultivars that are often used in these areas. Leaf biomass, relative leaf growth rate (RGR_L), concentration of B, nitrate (NO₃⁻), ammonium (NH₄⁺), organic N, amino acids and soluble proteins, as well as nitrate reductase (NR), nitrite reductase (NiR), glutamine synthase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH) activities were analysed in leaves. Boron toxicity significantly decreased leaf biomass, RGR_L, organic N, soluble proteins, and NR and NiR activities. The lowest NO₃⁻ and NH₄⁺ concentration in leaves was recorded when plants were supplied with 2.0 m m B in the root medium. Total B, amino acids, activities of GS, GOGAT and GDH increased under B toxicity. Data from the present study prove that B toxicity causes inhibition of NO₃⁻ reduction and increases NH₄⁺ assimilation in tomato plants.     

Effect of darkness and CO2 starvation on NH4+ and NO3− assimilation in the unicellular alga Cyanidium caldarium

Cyanidium caldarium (Tilden) Geitler, a non-vacuolate unicellular alga, resuspended in medium flushed with air enriched with 5% CO₂, assimilated NH₄⁺ at high rates both in the light and in the dark. The assimilation of NO₃⁻, by contrast, was inhibited by 63% in the dark. In cell suspensions flushed with CO₂-free air, NH₄⁺ assimilation decreased with time both in the light and in the dark and ceased almost completely after 90 min. The addition of CO₂ completely restored the capacity of the alga to assimilate NH₄⁺. NO₃⁻ assimilation, by contrast, was 33% higher in the absence of CO₂ and was linear with time. It is suggested that NO₃⁻ and NH₄⁺ metabolism in C. caldarium are differently controlled in response to the light and carbon conditions of the cell.     

Effects of osmolality and ions on the motility of stripped and testicular sperm of freshwater-and seawater-acclimated tilapia, Oreochromis mossambicus

A significantly higher concentration of testicular spermatozoa was obtained from freshwater Oreochromis mossambicus (9·9×10⁹ spermatozoa ml⁻¹) than seawater O. mossambicus (4·6×10⁹ spermatozoa ml⁻¹). The mean osmolality of the urine of freshwater fish (78·5 mOsmol kg⁻¹) was significantly different from that of seawater fish (304·8 mOsmol kg⁻¹). The mean length of the mid-piece of the spermatozoa together with the tail was more variable in freshwater O. mossambicus (8·80±0·23μm) than in seawater specimens (8·27±0·18 μm). Stripped sperm of freshwater O. mossambicus was highly contaminated by urine which was a good activator of sperm motility in O. mossambicus held in both fresh and sea water. The osmolality for initiation of motility in freshwater O. mossambicus spermatozoa was from 0 to 333 mOsmol kg⁻¹ while for seawater O. mossambicus spermatozoa it was from 0 to 1022 mOsmol kg⁻¹. The optimum osmolality for motility was from 70 to 333 mOsmol kg⁻¹ for freshwater O. mossambicus spermatozoa and from 333 to 645 mOsmol kg⁻¹ for seawater fish. In freshwater O. mossambicus spermatozoa, the presence of 20 mM CaCl₂ increased the permissive osmolality of NaCl from 184 to 645 mOsmol kg⁻¹. For seawater O. mossambicus spermatozoa, solutions of NaCl devoid of CaCl₂ were unable initiate motility, but the addition of 1·5 to 30 mM CaCl₂ to the NaCl solution (0–934 mOsmol kg₁) had a full motility initiating effect.     

Time-course of inorganic nitrogen uptake and incorporation by natural populations of freshwater phytoplankton

SUMMARY. 1. Time-course measurements of NH₄⁺ and NO₃⁻uptake were made on the natural phytoplankton populations in a eutrophic lake at a time when these nutrients were at their lowest annual concentration.
2. Both NH₄⁺ and NO₃⁻ uptake was increased at least five-fold during the first 5 min of incubation following near saturating pulses of these nutrients.
3. Elevated uptake was also observed following low level (∼2μg N 1⁻¹) pulses of NH₄⁺ and NO₃⁻, but substrate depletion during the first hour of incubation may have been partially responsible for this apparent enhancement.
4. Incorporation of ^I5N into TCA-insoluble material (protein) following the saturating NH₄⁺ pulse was increased less than total cellular ¹⁵N uptake, whereas no elevation of ¹⁵N incorporation into protein was observed following a saturating NO₃⁻pulse.
5. The percentage of ^I5N incorporated into protein, with respect to total cellular uptake, was ∼32% and ∼12% for NH₄⁺ and NO₃⁻, respectively, following 5 h of incubation.     

Elevated oxygen uptake and high rates of nitrogen excretion in early life stages of the cobia Rachycentron canadum (L.), a fast-growing subtropical fish

Physiological energetics of cobia Rachycentron canadum were quantified for 18 to 82 days post-hatch (dph) hatchery-reared juveniles to better understand energy transformation and its implications in growth and survival. Mean oxygen consumption rates ( ; mg O₂ h⁻¹) of fish fed ad libitum and fish that were starved significantly increased with increasing wet mass (M; g), = 1·4291 M ^0·8119 and = 1·1784 M ^0·7833, respectively, with a significant reduction in mean metabolic rates of starved fish (19 to 27% specific dynamic action; SDA). Total ammonia nitrogen excretion rates ( A _MM, μmol h⁻¹) also scaled with M and significantly decreased after starvation. Mean mass-specific A _MM and urea excretion rates are the highest reported in the literature, with urea accounting for approximately half the total nitrogen excretion measured in both fed and starved fish. Relatively high energetic rates may allow cobia to develop rapidly into pre-juveniles and be less susceptible to predation and starvation at a comparatively early age.     

An alternative explanation for the post-disturbance NO3– flush in some forest ecosystems

The appearance of soil NO₃^– after forest disturbance is commonly ascribed to a higher availability of NH₄⁺ to autotrophic nitrifiers, or to a reduction in available-C resulting in lower microbial assimilation of NO₃^–. Alternatively, it has been proposed that increasing NH₄⁺ pools following disturbance could increase net nitrification by reducing microbial assimilation of NO₃^–. Forest floor material was collected from shelterwood harvest plots which displayed both low available-C and low NH₄⁺ pools, and where previous experiments had suggested the prevalence of heterotrophic nitrification. Subsamples were amended with incremental rates of glucose-C or NH₄⁺, and gross NO₃^– transformation rates were measured by isotope dilution. Glucose-C additions had little effect on the net difference between gross NO₃^– production and consumption rates. On the other hand, NH₄⁺ additions caused gross NO₃^– consumption processes to decrease sharply, while gross NO₃^– production processes remained constant. The results suggest that NH₄⁺ can have an immediate positive effect on net nitrification rates by suppressing NO₃^– assimilation and uptake systems.     

Isolation, characterization, and nitrification potential of a methylotroph and two heterotrophic bacteria from a consortium showing methane-dependent nitrification

Abstract A methanotrophic nitrifying consortium was previously obtained from a humisol which showed CH₄-dependent nitrification. Although the methanotroph could not be obtained in pure culture, three other members of the consortium have been isolated: An obligately methylotrophic Methylobacillus (Is-1) which grows only on CH₃OH and does not nitrify; a Pseudomonas (Is-2) which grows on Is-1 culture filtrate and produces NO₂⁻, NO₃⁻ and N₂O from NH₂OH, and NO₃⁻ from NO₂⁻; and a second Pseudomonas (Is-3) which produces NO₃⁻ from NH₄⁺ or NO₂⁻, and N₂O from NH₂OH. A model is proposed for the trophic relations and nitrogen transformations in the consortium which may apply to some natural systems.     

A comparison of NH4+ and NO3– net fluxes along roots of rice and maize

Net fluxes of NH₄⁺ and NO₃^– along adventitious roots of rice ( Oryza sativa L.) and the primary seminal root of maize ( Zea mays L.) were investigated under nonperturbing conditions using ion-selective microelectrodes. The roots of rice contained a layer of sclerenchymatous fibres on the external side of the cortex, whereas this structure was absent in maize. Net uptake of NH₄⁺ was faster than that of NO₃^– at 1 mm behind the apex of both rice and maize roots when these ions were supplied together, each at 0·1 mol m^–3. In rice, NH₄⁺ net uptake declined in the more basal regions, whereas NO₃^– net uptake increased to a maximum at 21 mm behind the apex and then it also declined. Similar patterns of net uptake were observed when NH₄⁺ or NO₃^– was the sole nitrogen source, although the rates of NO₃^– net uptake were faster in the absence of NH₄⁺. In contrast to rice, rates of NH₄⁺ and NO₃^– net uptake in the more basal regions of maize roots were similar to those near the root apex. Hence, the layer of sclerenchymatous fibres may have limited ion absorption in the older regions of rice roots.     

Haloarchaeal assimilatory nitrate-reducing communities from a saline alkaline soil

Assimilatory nitrate reduction (ANR) is a pathway wherein NO₃⁻ is reduced to NH₄⁺, an N species that can be incorporated into the biomass. There is little information about the ANR genes in Archaea and most of the known information has been obtained from cultivable species. In this study, the diversity of the haloarchaeal assimilatory nitrate-reducing community was studied in an extreme saline alkaline soil of the former lake Texcoco (Mexico). Genes coding for the assimilatory nitrate reductase ( narB ) and the assimilatory nitrite reductase ( nirA ) were used as functional markers. Primers to amplify and detect partial narB and nirA were designed. The analysis of these amplicons by cloning and sequencing showed that the deduced protein fragments shared >45% identity with other NarB and NirA proteins from Euryarchaeota and <38% identity with other nitrate reductases from Bacteria and Crenarchaeota . Furthermore, these clone sequences were clustered within the class Halobacteria with strong support values in both constructed dendrograms, confirming that desired PCR products were obtained. The metabolic capacity to assimilate nitrate by these haloarchaea seems to be important given that at pH 10 and higher, NH₄⁺ is mostly converted to toxic and volatile NH₃, and NO₃⁻ becomes the preferable N source.     

Cytosolic and chloroplastic glutamine synthetase of sugarbeet (Beta vulgaris) respond differently to organ ontogeny and nitrogen source

Changes in the activity and subunit composition of cytosolic glutamine synthetase (GS 1; EC 6.3.1.2) and chloroplastic GS (GS 2) were studied in response to an internal (organ ontogeny) and external signal (N source: NO₃⁻ or NH₄⁺). Maximum GS 1 activity of all organs examined was measured in the fibre roots, irrespective of the N source. The response of GS 1 to the N source was, however, organ specific. In the fibre roots, NH₄⁺ nutrition resulted in a 2- to 7-fold (based on protein or freshweight, respectively) increase of GS 1 activity compared to NO₃⁻-grown plants. In contrast to the roots, GS 1 activity in the leaf blades was 2-fold lower with NH₄⁺ nutrition, whereas only minor changes occurred in the petioles. GS 2 activity was highest in the mature and senescing leaf blade; activity was 2-fold higher with NH₄⁺ than with NO₃⁻ nutrition. Not only activity, but also subunit composition of GS 1 changed during organ ontogeny as well as in response to the N source. In contrast to GS 1, only minor changes were evident in GS 2 subunit composition, despite significant changes in GS 2 activity. Up to 5 different GS 1 subunits of ≈41–43 kDa were separated; they were identical in all organs examined. GS 2 was composed of 4 different subunits of ≈48 kDa.     

Net fluxes and pools of nitrogenous compounds during suspension culture of photoautotrophic Chenopodium rubrum cells

Abstract. Suspension cultured cells of Chenopodium rubrum were grown photoautotrophically under a diurnal light-dark cycle of 16-8h. The following phases of the batch culture were differentiated: a short lag, a cell division phase terminated by a pronounced transition to stationary maintenance which finally gradually passed into senescence. Nitrogen fluxes typical of these stages were followed by measuring uptake of NO₃ and NH₄⁺ from the medium and their incorporation into the cellular fractions of nitrogenous compounds. Activities of seven N-metabolizing enzymes were determined. Compartmentation of enzymes and nitrogenous compounds was analysed after isolation of intact chloroplasts and vacuoles from protoplasts. Eighty-two per cent of the N originally present in the medium was taken up and incorporated to an extent of 80% into protein until the end of the division phase. Net protein synthesis ceased upon transition to the stationary phase. During the division phase a vacuolar pool of NO₃ was established and then maintained throughout the resting phase. Free cellular NH₄⁺ was not localized within the vacuole and responded to the ammonium content of the medium. Amino acids accumulated in the cells especially during the stationary phase, during which they were present in the vacuole. Typical nitrogen relations are portrayed as flux diagrams for one day of each of the essential developmental phases. The enzyme activities were easily sufficient to account for the observed flow rates of the corresponding nitrogenous compounds. Hence, uptake of NO₃ and NH₄⁺ must be considered as steps limiting N metabolism in Chenopodium rubrum cell suspensions.     

Photosynthate costs associated with the utilization of different nitrogen–forms: influence on the carbon balance of plants and shoot–root biomass partitioning

The photosynthate costs of processes (amino acid and protein synthesis and turnover, and pH regulation) associated with the utilization of nitrate (NO₃⁻), ammonium (NH₄⁺) or glutamine (Gln) for plant growth were estimated. Based on these estimates, the effects of these forms of nitrogen (N) on the carbon balance of plants and on shoot–root biomass allocation were evaluated. The results indicated that NO₃⁻ as an N source for plant growth is not substantially more expensive to utilize than either NH₄⁺ or Gln, particularly in the long term when costs due to protein turnover dominate the total costs of N utilization. It is also suggested that the photosynthate use in processes associated with N assimilation has little impact on the carbon balance of plants, and hence on shoot–root biomass allocation.