Nitrate reductase activity in Zostera marina

Eelgrass (Zostera marina L.) has access to nutrient pools in both the water column and sediments. We investigated the potential for eelgrass to utilize nitrate nitrogen by measuring nitrate reductase (NR) activity with an in vivo tissue assay. Optimal incubation media contained 60 mM nitrate, 100 mM phosphate, and 0.5% 1-propanol at pH 7.0. Leaves had significantly higher NR activity than roots (350 vs 50 nmoles NO ₂ ^– produced g FW^–1 h^–1). The effects of growing depth (0.8 m MLW, 1.2 m, 3.0 m, 5.0 m) and location within the eelgrass meadow (patch edge vs middle) on NR activity were examined using plants collected from three locations in the Woods Hole area, Massachusetts, USA, in July 1987. Neither depth nor position within the meadow appear to affect NR activity. Nitrate enrichment experiments (200 M NO ₃ ^– for 6 d) were conducted in the laboratory to determine if NR activity could be induced. Certain plants from shallow depth (1.2 m) showed a significant response to enrichment, with NR activity increasing from >100 up to 950 nmoles NO ₂ ^– g FW^–1 h^–1 over 6 d. It appears that Z. marina growing in very shallow water (0.8 m) near a shoreline may be affected by ground water or surface run-off enrichments, since plants from this area exhibited rates up to 1 600 nmol NO ₂ ^– g FW^–1 h^–1. Water samples from this location consistently had slightly higher NO ₃ ^– concentrations (1.4 M) than all other collection sites (0.7 M). Thus, it is possible that chronic run-off or localized groundwater inputs can create sufficient NO ₃ ^– enrichment in the water column to induce nitrate reductase activity in Zostera leaves.     

Different patterns of growth and nitrogen uptake in two clones of marineSynechococcus spp.

We grew marineSynechococcus Clones WH7803 and WH8018 at 150µE m^–2 s^–1 in dilute batch cultures with NH ₄ ⁺ as the limiting nutrient. The maximal uptake capacities for NH ₄ ⁺ and NO ₃ ^- were measured in frequent experiments during log and stationary phases of growth. Clone WH7803, originally isolated from oceanic waters, had a specific uptake rate of NH ₄ ⁺ that approximated the maximum (log phase) specific growth rate (ca ~ 0.025 h^–1). NO ₃ ^- uptake was observed only after nitrogen in the media was depleted; the NO ₃ ^– uptake capacity was ca 12% the capacity for NH ₄ ⁺ uptake throughout the nitrogen depleted period. Growth was arrested upon nitrogen depletion, but resumed soon after reinoculation into fresh media, even after 5 d of starvation. Clone WH8018, originally isolated from coastal waters, revealed a five-fold enhancement in the NH ₄ ⁺ uptake rate relative to growth rate at the time of nitrogen depletion. As nitrogen starvation proceeded, this enhancement was reduced. This clone, too, was able to take up NO ₃ ^- once nitrogen in the media was depleted, but only after ca 20 h. Growth continued for a limited period during nitrogen depletion, but nitrogen-starved cells were slow to recover upon reinoculation into fresh media. We speculate that clonal differences may reflect differences in the molecular regulation of nitrogen assimilation.     

Nitrogen supply,tissue composition and frond growth rates for Macrocystis pyrifera off the coast of southern California

Seasonal changes in ambient NO ₃ ^– and NH ₄ ⁺ , tissue composition (N, C, and C/N ratio), and frond growth rates for Macrocystis pyrifera (L.) Agardh were examined. Ambient NO ₃ ^– showed distinct seasonal variations. Frond growth rates were variable, but showed no clear correlation to ambient NO ₃ ^– . The average N content of plant tissue did, however, show the same seasonal variations as ambient NO ₃ ^– . The longitudinal distribution of total tissue N and various components of tissue N along fronds were also analyzed. Several distinct patterns were found: high levels of protein N at growing tips and elevated levels of soluble N in lower parts of the frond. Free amino acids accounted for a major portion of the soluble N, but neither NO ₃ ^– nor NH ₄ ⁺ accumulated in the plant tissue. The longitudinal distribution of N along the fronds is compared to reported variations in C metabolism, and it is concluded that C and N sourcesink relations do not always coincide and bidirectional translocation may occur.     

Nitrate reductase and urease enzyme activity in the marine diatom<Emphasis Type="Italic"> Thalassiosira weissflogii</Emphasis> (Bacillariophyceae): interactions among nitrogen substrates

The dissolved nitrogen pool in aquatic systems is comprised of many different nitrogen forms, both inorganic and organic. Interaction among these nitrogen forms at the level of uptake and enzyme activity is, with the exception of NH₄⁺ and NO₃^–, not completely understood. Nitrate reductase (NR) and urease (UA) activities in the marine diatom Thalassiosira weissflogii (Grunow) Fryxell et Hasle were measured in NO₃^–, NH₄⁺, and urea-sufficient cultures before and after challenge additions of NH₄⁺, NO₃^–, and urea in a factorial design. NR and UA were constitutively expressed during growth on NO₃^–, NH₄⁺, and urea. Growth on NH₄⁺ or urea resulted in NR activities that were <10% of the activity observed in the NO₃^–-grown culture, while growth on NO₃^– resulted in UA values that were ~35% of the activities during growth on either NH₄⁺ or urea. The addition of NH₄⁺ or urea to NO₃^–-grown cultures resulted in an immediate decrease in cellular NO₃^– uptake rate, which was not mirrored by an immediate repression of in vitro NR activity; however, the diel peak in NR was suppressed in these challenge experiments. The addition of NO₃^– or NH₄⁺ to urea-grown cultures resulted in non-significant decreases in the urea uptake rate. UA was not impacted by NO₃^– addition, but NH₄⁺ addition significantly decreased UA throughout the experiment. These studies demonstrate that the uptake and assimilation of NO₃^– and urea may not be subject to the same internal feedback mechanism when challenged with other nitrogen substrates.Communicated by J.P. Grassle, New Brunswick     

Relative growth of different species of marine algae in wasterwater-seawater mixtures

In recent studies, we developed a combined nutrient removal-marine aquaculture process for the tertiary treatment of wastewater and the production of commercially important shellfish. Part of this process consists of an outdoor mass cultivation system for marine algae. During our previous experiments we observed that marine diatoms almost exclusively are the dominant algal species in our outdoor cultures. To better understand this phenomenon of diatom dominance we grew 16 species of marine algae in continuous monoculture under laboratory conditions simulating to some degree the conditions prevailing in our outdoor experiments. Species such as Skeletonema costatum, Monochrysis lutheri and Tetraselmis sp., which were never dominant in our outdoor cultures, grew as well in monoculture, as Phaeodactylum tricornutum, frequently, the prevalent species outdoors. However, when monocultures of Dunaliella tertiolecta and Thalassiosira pseudonana (3H) were purposely contaminated with P. tricornutum, the latter species quickly became dominant. It is suggested that a complex interaction of environmental factors is usually responsible for the dominance of a particular species; one such factor may be the nitrogen source in the growth media. Under conditions of virtually, complete nitrogen assimilation, the carbon: nitrogen ratio in the algae was high (7 to 8) when NO ₃ ^- –N was the source of nitrogen, and low (4 to 6) when NH ₄ ⁺ –N was the prime form of nitrogen. When algal growth was low, resulting in a large inorganic nitrogen residue, the carbon:nitrogen ratio was low regardless of whether NO ₃ ^- –N or NH ₄ ⁺ –N was the main nitrogen source.Contribution No. 3297 from the Woods Hole Oceanographic Institution.     

Comparisons of nutrient uptake rates for Baltic macroalgae with different thallus morphologies

In-situ experiments were performed during different seasons to determine uptake rates of PO _3- ⁴ , NH ₊ ⁴ and NO _- ³ within ecologically representative ranges of nutrient concentrations, of dominant macroalgae in the Baltic Sea. Uptake rates were governed by nutrient concentrations, water temperature and thallus morphology, but not by the phylogenetic affinity of the species. Nitrogen uptake rates were always higher than those of phosphorus at the same concentrations, and NH ₊ ⁴ –N uptake rates exceeded those of NO _- ³ –N. The lowest uptake rates occurred among the late successional, long-lived, coarse species with low surface: volume ratios (Fucus vesiculosus, Furcellaria lumbricalis andPhyllophora truncata). The highest uptake rates were measured for short-lived, opportunistic algae, filamentous or with numerous hairs, (Cladophora glomerata, Enteromorpha ahlneriana, Scytosiphon lomentaria, Dictyosiphon foeniculaceus andCeramium tenuicorne). The latter group also had the highest V_max:k_max ratios, which indicates a more competitive advantage for nutrient uptake at low concentrations.     

Ammonium regeneration by microzooplankton in the Oslofjord

Concentrations of dissolved inorganic nitrogen compounds above the pycnocline in the Oslofjord are very low in the summer, with turnover times of the inorganic N pools of no more than a few hours. To investigate the possibility that continued phytoplankton growth in the summer depends on ammonium excretion by microzooplankton, rates of NH ₄ ⁺ regeneration and assimilation were measured by a ¹⁵N isotope dilution method. Daytime regeneration rates at 0–2 m depth were 0–28% of the calculated assimilation rates at ambient NH ₄ ⁺ concentrations. Regeneration was faster during a dinoflagellate bloom in August than in mixed diatom-dinoflagellate blooms in June and September. Most of the NH ₄ ⁺ appeared to be produced by juvenile copepods, rotifers, tintinnids, and heterotrophic dinoflagellates in the size fraction 45–200 m.     

Effect of algal bloom deposition on sediment respiration and fluxes

Using sediment cores collected in November 1989 from Aarhus Bight, Denmark, the fluxes of O₂, CO₂ (total CO₂), NH ₄ ⁺ , NO ₃ ^– +NO ₂ ^– and DON (dissolved organic nitrogen) across the sediment-water interface were followed for 20 d in an experimental continous flow system. On day 7, phytoplankton was added to the sediment surface, to see the result of simulated algal bloom sedimentation. Benthic O₂ consumption and CO₂ efflux, 38 to 41 mmol O₂ m^-2 d^-1 and 25 to 30 mmol CO₂ m^-2 d^-1, respectively, immediately increased by 39% and 100% after the algal addition, but gradually declined to the orginal level. Fluxes of NH ₄ ⁺ (1.0 to 1.2 mmol m^-2 d^-1) and DON (0.3 to 0.9 mmol m^-2 d^-1) increased due to the organic substrate addition. NH ₄ ⁺ and NO ₃ ^– flux changed direction, becoming an efflux and influx, respectively, for a few days and a large amount of DON (max. rate 4.0 mmol m^-2 d^-1) was immediately produced either by bacterial hydrolytic activity or from autolysis of the algae. DON was the most significant nitrogen component in pore water and in terms of N-flux from sediment. A temporary stimulation of anaerobic respiration processes (sulfate reduction and denitrification) and a decrease in nitrification were indicated. After the effect of the organic addition had declined, the fluxes gradually reverted to the original rates. The halflife of the added algal material, of which 20 to 30% was very labile, was estimated to be 2 to 3 wk.     

Inorganic nitrogen metabolism inUlva rigida illuminated with blue light

Inorganic nitrogen metabolism in blue light was studied for the green algaUlva rigida C. Agardh collected in the south of Spain (Punta Carnero, Algeciras) in the winter of 1987. NH₄ ⁺ has been reported to inhibit NO₃ ^- uptake; however,U. rigida showed a net NO₃ ^- uptake even when the NH₄ ⁺ concentration of the external medium was three or four times greater than the concentration of NO₃ ^-. NO₃ ^- uptake rates were similar in both darkness and in blue light of various photon fluence rates (PFR) ranging from 17 to 160 mol m^-2 s^-1. Since NO₃ ^- uptake is an active mechanism involving the consumption of ATP, respiratory metabolism can provide enough ATP to maintain the energetic requirement of NO₃ ^- transport even in darkness. In contrast, NO₃ ^- reduction inU. rigida was highly dependent on the net photosynthetic rate. After 7 h in blue light, intracellular NO₃ ^- concentrations ([NO₃ ^-] _i ) were higher in specimens exposed to intensities below the light compensation point (LCP) than in those incubated at a PFR above the LCP. When PFR is below the light compensation point, NO₃ ^- reduction is low, probably because all the NADH produced by the cells is oxidized in the respiratory chain in order to produce ATP to maintain a steady NO₃ ^- transport rate. The total nitrogen (TN) and carbon (TC) contents decreased from darkness to 33 mol m^-2 s^-1 in blue light. In this range, catabolic processes prevailed over anabolic ones. In contrast, increases in TN and TC contents were observed above the light compensation point. The C : N ratio increased with light intensity, reaching a stable value of 17 at 78 mol m^-2 s^-1 in blue light. Intracellular NO₃ ^- concentration and NO₃ ^- reduction appear to be directly controlled by light intensity. This external control of [NO₃ ^-]_i and the small capacity ofU. rigida to retain incorporated NO₃ ^-, NO₂ ^- and NH₄ ⁺ ions may explain its nitrophilic character.     

Effect of ammonia on survival and osmoregulation in different life stages of the lobsterHomarus americanus

Acute toxicity of ammonia was determined for cultured larval, postlarval, and wild adult lobsters (Homarus americanus) in 1988. Ammonia tolerance was found to increase with ontogenetic development. Based on 96-h LC₅₀ values of 58 mg l^–1 NH₄ ⁺ + NH₃ l^–1 seawater (0.72 mg NH₃ l^–1) for Stage I larvae, 87 mg NH₄ ⁺ + NH₃ l^–1 (1.7 mg NH₃ l^–1) for Stage II larvae, 125 mg NH₄ ⁺ + NH₃ l^–1 (2.13 mg NH₃) for Stage III larvae, 144 mg NH₄ ⁺ + NH₃ l^–1 (2.36 mg NH₃ l^–1) for Stage IV postlarvae, 377 mg NH₄ ⁺ + NH₃ l^–1 (5.12 mg NH₃ l^–1) for adult lobsters at 5°C and 219 mg NH₄ ⁺ + NH₃ l^–1 (3.25 mg NH₃ l^–1) for adult lobsters at 20°C, recommendations for safe levels of total ammonia and un-ionized ammonia were calculated using an application factor of 0.1. Effects of ammonia on osmoregulatory capacity were studied on postlarvae and adults. Ability of postlarvae and adults to hyper-regulate in low-salinity media decreased after exposure to ammonia. In postlarval lobsters, osmoregulatory capacity was significantly affected in ammonia concentrations exceeding 32 mg l^–1. Osmoregulatory capacity in adult lobsters (5 and 20°C) was affected at 150 mg l^–1. In postlarval lobsters, a minimum exposure time of 12 h was required to impair osmoregulatory capacity. The decrease in hemolymph osmotic pressure was caused by lower hemolymph sodium concentrations. The presence of ammonia in the external medium could markedly affect the Na⁺/NH₄ ⁺ transport mechanism by permanently, temporarily, or partially impairing the transport sites for sodium.     

Ingestion-independent rates of ammonium excretion by the copepod Calanus pacificus

The relationship between food ingested and NH ₊ ⁴ excretion rate was investigated for female Calanus pacificus collected in August, 1982, from the San Juan Archipelago, Washington State, USA. The copepods were preconditioned to 6 densities of the diatom Thalassiosira weissflogii (0 to 10⁴ cells ml^–1) for 30 h before the experiment. The experiment was conducted with nutrients added in excess to maintain equal rates of NH ₊ ⁴ uptake by the diatoms at all densities. Although ingestion rates of C. pacificus varied from 0 to over 20% of body N d^–1 at the different food levels, excretion was a constant 6.6 nM NH ₊ ⁴ copepod^–1 h^–1 or about 10% of body N d^–1. This ingestion-excretion relationship, which is consistent with previous respiration and fecundity studies, suggests that the ecological dominance of C. pacificus only under conditions of high food abundance may be due to a dramatic increase in its growth efficiency as ingestion increases above the level supporting a constant metabolic rate. The maintenance of a constant level of metabolism during relatively short periods of low food abundance may be advantageous if it allows the copepod to exploit more effectively short-term variability in its food resulting from environmental heterogeneity or vertical migration.Contribution No. 1360 from the School of Oceanography, University of Washington, Seattle, Washington 98195, USA     

Effects of methionine sulfoximine on growth and nitrogen assimilation of the marine microalga Chlorella autotrophica

Chlorella autotrophica Shihira and Krauss (clone 580), a euryhaline microalga from the marine coastal environment is subject to large fluctuations in external salinity and nitrogen supply. The alga exhibits maximum growth at salinities lower than 100% ASW (artificial seawater). Cells divide faster and show higher cell yields when the supply of either NH ₄ ⁺ or NO ₃ ^- is increased above 0.2 mM. Cells growing on NH ₄ ⁺ show high levels of NADPH-glutamate dehydrogenase (GDH) activity, and the levels of glutamine synthetase (GS) are decreased to very low levels under these conditions. Methionine sulfoximine (MSX), an inhibitor of GS, has little effect on cell division and nitrogen assimilation of cells growing on NH ₄ ⁺ . Cells growing on NO ₃ ^- , however, show marked inhibition (65%) in nitrogen assimilation in the presence of 5 mM MSX. This MSX concentration also causes growth retardation and a progressive decrease in cell protein and nitrogen content. GS is almost completely inhibited by 5 mM MSX in both NH ₄ ⁺ and NO ₃ ^- -grown cells. Cells growing on NH ₄ ⁺ maintain high levels of NADPH-GDH activity in the presence of MSX. NADPH-GDH activity in MSX-treated NO ₃ ^- -grown cells increases, and, in the presence of 5 mM MSX, reaches 40% of the level found in NH ₄ ⁺ -grown cells. These results are consistent with NADPH-GDH providing an alternate pathway for NH ₄ ⁺ assimilation by this marine Chlorella species.     

Photosynthetic acclimation and biochemical responses ofGelidium sesquipedale cultured in chemostats under different qualities of light

The red algaGelidium sesquipedale (Clem.) Born. et Thur. has been cultured in chemostats to assess the effects of light quality and photon-fluence rate (PFR) on growth, photosynthesis and biochemical composition. Plants under blue and red light (BL and RL) showed higher growth rates than under white light (WL) of the same PFR (40 mol m^–2 s^–1). The light-saturated rate of photosynthesis was higher for algae grown under BL and RL than for algae grown under WL. When algae were transferred to WL of moderate PFR (100 mol m^–2 s^–1), the light-saturated rate of photosynthesis decreased, being higher in previously RL-grown algae than in previously BL- and WL-grown algae. The initial slope of photosynthesis-irradiance (PI) curves () was affected by PFR but not by light quality. Pigment content was little affected by light quality. Light-quality treatments also affected the biochemical composition of the alga; previous exposure to various light treatments activate or repress several metabolical pathways that are fully expressed in the subsequent phase of WL of moderate PFR. Thus, phycobiliproteins and soluble proteins increased for previously BL- and RL-grown algae, whereas insoluble carbohydrate concentration was reduced, indicating a change of the C-partitioning between carbon compounds and organic nitrogen compounds. Inorganic nitrogen metabolism was also affected by light: under WL of moderate PFR, NO₃ ^– was totally depleted from sea water, and maximal values of NO₃ ^– uptake were recorded. In addition, neither NO₂ ^– nor NH₄ ⁺ was released. However, when algae were transferred to a low PFR, there was a drastic reduction of NO₃ ^– uptake under WL, which only partially recovered over time. It was accompanied by the release of NO₂ ^–, but not NH₄ ⁺, to the culture medium. Under BL and RL, however, there was a transient enhancement of NO₃ ^– uptake that was followed by a net release of NO₂ ^– and NH₄ ^–. Growth rates were not correlated with PFR. This could be due to the the dynamics of internal carbon mobilization and accumulation in the algae. When algae were exposed to a moderate PFR of WL, carbon requirements for growth were satisfied by photosynthesis. Thus, there was a net accumulation of carbon in the tissue. In contrast, when algae were exposed to low PFRs of either WL, BL or RL, observed growth rates could not be maintained by photosynthesis and carbon was mobilized.     

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     

Growth pattern andβ-dimethylsulphoniopropionate (DMSP) content of green macroalgae at different irradiances

Growth rates and intracellular-dimethylsulphoniopropionate (DMSP) concentrations of five green algal species collected from different geographic regions in 1986 and 1989 were determined under four photon flux rates. InUlothrix implexa, U. subflaccida andAcrosiphonia arcta from Antarctica, growth was light-saturated at lower irradiances than in temperateUlva rigida from Southern Chile andBlidingia minima from Germany. The DMSP content ofUlothrix implexa, A. arcta andUlva rigida was directly correlated with the light factor: with increasing irradiance, algal DMSP level increased. In contrast, inUlothrix subflaccida andB. minima DMSP concentrations gradually decreased up to a photon flux rate of 30µmol m^–2 s^–1, then increased markedly under the highest photon flux rate tested. In non-growing, dark-incubatedA. arcta DMSP content was reduced by 35%, while the DMSP pool of all other species remained unchanged, at the level of pre-culture conditions. Under full darkness all plants exhibited a significantly higher DMSP concentration compared with algae grown at low photon flux rates of 2 to 30µmol m^–2 s^–1. These data show a correlation between growth pattern and DMSP biosynthesis, and may point to a species-specific minimum amount of light energy necessary for DMSP accumulation.Contribution no. 302 of the Alfred Wegener Institute of Polar and Marine Research     

The effects on human health and hydrogeochemical characteristics of the Kirkgeçit and Ozancik Hot Springs,Canakkale, Turkey

This investigation was carried out to determine the hydrogeochemical characteristics of the Kirkgeçit and Ozancik hot springs. The study areas are located northeast and southwest of the town of Çan, Çanakkale. During the investigation, geological maps of the hot springs and its surroundings were prepared, and hot waters and rock samples were collected from the study sites. The Paleogene–Neogene aged andesite, trachyandesite, andesitic tuff, silicified tuff and tuffites form the basement rocks in the Ozancik hot spring area. In the Kirkgeçit hot spring area, there are Lower Triassic aged mica and quartz schists at the basement rocks. The unit is covered by limestones and marbles of the same age. They are overlain by Quaternary alluvial deposits. A chemical analysis of the Kirkgeçit hot water indicates that it is rich in SO₄ ^2– (1200.2 mg L^–1), Cl^– (121.7 mg L^–1), HCO₃ ^– (32.5 mg L^–1), Na⁺ (494 mg L^–1), K⁺ (30.2 mg L^–1), Ca²⁺ (102 mg L^–1), Mg²⁺ (15.2 mg L^–1), and SiO₂ (65.22 mg L^–1). Chemical analysis of the Ozancik hot water indicates that it is rich in SO₄ ^2– (575 mg L^–1), Cl^– (193.2 mg L^–1), HCO₃ ^– (98.5 mg L^–1), Na⁺ (315 mg L^–1), K⁺(7.248 mg L^–1), Ca²⁺ (103 mg L^–1), Mg²⁺ (0.274 mg L^–1), and SiO₂(43.20 mg L^–1). The distribution of ions in the hot waters on the Schoeller diagram has an arrangement of r(Na⁺+K⁺)>rCa²⁺>rMg²⁺ and r(SO₄ ^2–)>rCl^–>r(HCO₃ ^–). In addition, the inclusion of Fe²⁺, Cu²⁺, Cr³⁺, Mn²⁺, Ni²⁺ and Hg²⁺ in the hot water samples indicates potential natural inorganic contamination. The water analysis carried out following the ICPMS-200 technique was evaluated according to the World Health Organisation and Turkish Standards. The use and the effects of the hot water on human health are also discussed in the paper.     

Effects of ammonia on survival and osmoregulation of various development stages of the shrimp Penaeus japonicus

In Penaeus japonicus, the tolerance to ammonia increased with the development from nauplius to late juvenile. The 48-h LC₅₀ of ammonia in nauplii (III–V), 96-h LC₅₀ in zoeae (I–III), mysis (I–III), post-larvae (PL1) and late juveniles (10.4±1.1 g) were respectively 5.0, 6.1 to 8.1, 9.4 to 10.9, 15.5 and 52.7 mg Nl^-1 (0.5, 0.6 to 0.7, 0.9, 1.3 and 3.1 mg NH₃–Nl^-1). In a chronic experiment (20 d), the LC₅₀ in post-larvae (PL1) was 19.1 (1.4) at 96 h and 16.2 mg Nl^-1 (1.3 mg NH₃–Nl^-1) at 480 h. Osmoregulatory capacity (OC) was calculated as the osmotic gradient between the hemolymph and the external medium at given salinities. The effects of ammonia on OC, Na⁺ and Cl^- regulation and gill Na⁺–K⁺ ATPase activity in late juveniles were examined in fullstrength seawater, SW (1050 mosm kg^-1, 36 S) and in dilute SW (450 mosm kg^-1, 15%.), after 48 or 96 h exposure to various concentrations of ammonia. Ambient ammonia disrupted both hypo- and hyper-osmoregulation; decreased OC resulted from impaired Na⁺ and Cl^- regulation. Gill Na⁺–K⁺ ATPase activity increased in SW and was not affected in dilute SW. The decrease of OC was ammonia-dose-dependent. The threshold ammonia concentrations affecting hypo-OC and hyper-OC were, respectively, 16 (1.3) and 32 mg Nl^-1 (2.3 NH₃–Nl^-1) for a 48 h exposure; these concentrations were lower than the 48-h LC₅₀ value, 65.3 mg Nl^-1 (3.5 NH₃–Nl^-1). The time course of exposure to sublethal ammonia (48 mg Nl^-1) demonstrated that the effect on osmoregulation was time-dependent. This effect was also temporary, and the exposed shrimps recovered control OC values after removal of excessive ambient ammonia. The possibility of using OC as an indicator of physiological condition in osmoregulating crustaceans and the acting mode of ammonia on osmotic and ionic regulation are discussed.     

Bioenergetics of fishes in a high-energy surf-zone

Energy budgets are proposed for four teleost and two elasmobranch species as well as for the main icthyofauna groups for a surf-zone ecosystem. The ecology of surf-zone fishes of eastern Cape beaches, Algoa Bay South Africa, is reviewed. Using the equationC=F+U+R _d +R _R +B, the following general energy budgets were derived for fishes: teleosts – 100=10+4+21+23+42; elasmobranchs – 100=11+2+16+24+48; whereC: food consumption;F: faeces;U: nonfaecal excretion;R _d : apparent specific dynamic action;R _R : routine metabolism;B: growth. These show that most of the energy consumed is used in metabolism (R _d +R _R ) and growth (B) whereas excretion (U) only accounts for a small portion. The energy budgets developed are within ranges recorded for other species. The main feeding groups of surf-zone icthyofauna are the southern mulletLiza richardsonii, the sandsharkRhinobatos annulatus, benthic feeders, zooplankton feeders, omnivorous and piscivorous fish with biomass values of 1000, 1000, 3000, 2400, 400 and 400 kJ m^–1, respectively; and annual consumption budgets of 22107, 13725, 65710, 65476, 9758 and 8517 kJ m^–1 yr^–1, respectively.L. richardsonii feeds mainly on surf diatoms, consuming 0.5% of total diatom production. Zooplankton production supplies 91%, and macrobenthic production 9%, of the energy needs of other non-piscivorous carnivorous fishes. Piscivorous fishes consume 30% of the available fish production. Nonfaecal-energy production (8229 kJ m^–1 yr^–1) is utilised by surf diatoms, and faecal-energy production (30 341 kJ m^–1 yr^–1), is returned to the detritus pool to be utilised by the microbial loop in surf-waters. Our current knowledge of surf-zone energetics indicates that fishes are important predators. This study confirms the concept that the ecosystem generates adequate food for the fish assemblage. Fishes recycle energy, as excretory products, via the detritus pool and surf-diatoms, while fishes moving across the outer boundary of the surf-zone export energy from the system. Data presented, therefore, also support the general concept of a self-sustaining beach/surf-zone ecosystem.Please address all correspondence and requests for reprints to Dr Du Preez at his present address: Research Unit for Fish Biology, Rand Afrikaans University, P.O. Box 524, Johannesburg 2000, Republic of South Africa     

Nitrogen Export from an Agriculture Watershed in the Taihu Lake Area, China

Temporal changes in nitrogen concentrations and stream discharge, as well as sediment and nitrogen losses from erosion plots with different land uses, were studied in an agricultural watershed in the Taihu Lake area in eastern China. The highest overland runoff loads and nitrogen losses were measured under the upland at a convergent footslope. Much higher runoff, sediment and nitrogen losses were observed under upland cropping and vegetable fields than that under chestnut orchard and bamboo forest. Sediment associated nitrogen losses accounted for 8–43.5% of total nitrogen export via overland runoff. N lost in dissolved inorganic nitrogen forms (NO ₃ ^– -N + NH ₄ ⁺ -N) accounted for less than 50% of total water associated nitrogen export. Agricultural practices and weather-driven fluctuation in discharge were main reasons for the temporal variations in nutrient losses via stream discharge. Significant correlation between the total nitrogen concentration and stream discharge load was observed. Simple regression models could give satisfactory results for prediction of the total nitrogen concentrations in stream water and can be used for better quantifying nitrogen losses from arable land. Nitrogen losses from the studied watershed via stream discharge during rice season in the year 2002 were estimated to be 10.5 kg N/ha using these simple models.     

A study on the landfill leachate and its impact on the groundwater quality of the greater area

Characterisation of the leachate originating from the Ano Liosia landfill (situated in Attica region, Greece) as well as assessment on the quality of the local aquifer were carried out. The experimental results showed that most of the parameters examined in the leachate samples such as colour, conductivity, TS, COD, NH₃–N, PO₄–P, SO₄ ^2–, Cl^–, K⁺, Fe and Pb were found in high levels. The organic load was quite high since the COD concentrations were in the range of 3250–6125mgL^–1. In addition, the low BOD/COD ratio (0.096–0.195), confirmed that the majority of this organic matter is not easily biodegradable. The groundwater near the landfill site was characterised as not potable and not suitable for irrigation water, since most of the physical and chemical parameters examined – such as colour, conductivity, DS, hardness, Cl^–, NH₃–N, COD, K⁺, Na⁺, Ca²⁺, Fe, Ni and Pb exceeded the permissible limits given by EE, EPA and the Greek Ministry of Agriculture. Furthermore, this study presents the application of the hydrologic evaluation of landfill performance (HELP) model for the determination of the yearly leakage from the base of the landfill after the final capping.