Impact of nitrogen cycling on stream water quality in a basin associated with forest, grassland, and animal husbandry, Hokkaido, Japan

The direct discharge of wastes from agricultural fields and livestock feedlots increases the concentration of nitrogen (N) in streams. This study was conducted to determine the impact of farm N budgets on stream water quality. In 1999–2000, we investigated an experimental livestock farm of 457 ha in the Kepau River watershed in Shizunai, Southern Hokkaido, Japan, where grasslands and maize fields account for 33% of the farm's total area. Annual N flow was calculated on the basis of the farm's land management records. Livestock was supplied with 15.2 t N yr⁻¹ from agricultural lands, which made the farm 81% self-sufficient. Livestock excreta produced 17.2 t N yr⁻¹, of which 4 t N yr⁻¹ was lost, probably by ammonia volatilization during decomposition. Apart from manure, the major N inputs were 9.1 t N yr⁻¹ of chemical fertilizers, 6.4 t N yr⁻¹ of atmospheric deposition, and 12.6 t N yr⁻¹ biological N fixation. The major outputs were uptake by forest vegetation of 11.0 t N yr⁻¹, denitrification of 1.5 t N yr⁻¹, and livestock feed production. Consequently, the annual surplus N on the whole farm was estimated to be 12.7 t N yr⁻¹, which corresponds to 28 kg N ha⁻¹ of agricultural land.The annual N load from the farm to the Kepau River was measured at 14.4 t N yr⁻¹. Ninety percent of this load, however, occurred during rainfall and spring snowmelt. Within one 2-week snowmelt period, 5.0 t N was discharged, which corresponds to 35% of the annual load. Although the average N concentration of stream water below the farm was 2.8 mg N L⁻¹, the maximum concentration recorded during the snowmelt season was 13.5 mg N L⁻¹. The N concentration of the stream water increased and the silica (Si) concentration decreased as the stream flow rate increased. Consequently, the molar ratio of Si/N frequently dropped below 2.7, the critical level for the occurrence of eutrophication. The large N load during rainfall and snowmelt could be ascribed to open ditches, which collect tile drainage and surface runoff from the fields, discharging it directly to the river, bypassing the forested riparian zone.     

Ammonium excretion by a mutant of the nitrogen-fixing cyanobacterium Anabaena siamensis

Anabaena siamensis isolated from rice fields in Thailand is a fast growing cyanobacterium with a high nitrogen-fixing activity. Mutant strains resistant to the l-glutamate analogue, l-methionine sulfoximine (MSX) were isolated by ethyl methanesulfonate mutagenesis. A stable mutant named A. siamensis SS1, which released ammonium to the medium, was studied further. In batch cultures the rate of ammonium production peaked at the early log phase and gradually decreased until the 4th day of growth when the cultures reached a density of 90 μg chl ml⁻¹. To obtain constant release of ammonium by SS1, continuous culture experiments were performed at a cell density of 5 μg chl ml⁻¹ and the following results were obtained: (1) growth rate as the parent (μ:0·123 h⁻¹) in the presence and absence of 500 μm MSX; (2) 48% GS transferase activity when compared with the parent; (3) ammonium excretion at a rate of 8 μmol (mg chl)⁻¹ h⁻¹ as measured up to 20 generations (120 h); (4) depressed nitrogenase activity; and (5) 30% higher nitrogenase activity than that of the parent. SS1 immobilized in alginate beads (5 μg chl ml⁻¹) exhibited values of glutamine synthetase and nitrogenase activity similar to those of free cells. However, ammonium excretion at the rate of 11·61 μmol (mg chl)⁻¹ h⁻¹ was obtained only up to 20 h after loading in bioreactors, due to the fast growth of SS1 as also occurred in batch cultures.     

Uptake and accumulation of zinc in juvenile rainbow trout,Salmo gairdneri

Synopsis The toxicity of zinc to rainbow trout was determined and the 72 h median lethal concentration was found to be 2.00 mg l^–1 in freshwater, hardness 7.50 mg l^–1 as calcium. An insignificant increase in zinc concentration of internal tissues occurred in fish exposed to 1.52 mg l^–1 in freshwater for 72 h. However, there was a significant uptake of zinc by gills and the body surface. Fish exposed to 10 mg l^–1 zinc for 72 h in two-thirds sea water showed significant zinc uptake by liver, rectum and muscle, when compared to control fish. Drinking rate decreased from 1.43 to 0.26 ml kg^–1 h^–1 when zinc sulphate was added to freshwater. Trout adapted to two-thirds sea water showed no decrease in drinking, about 7 ml kg^–1 h^–1 when zinc was added to the water.     

Na+ uptake through the brush border membranes of intestine from fresh water and sea-water adapted trout (Salmo gairdneri,R.)

Summary A comparative study of the mechanisms of Na⁺ absorption through brush border membranes of enterocytes from freshwater (FW) and seawater (SW) adapted trout were carried out using purified vesicle preparations. In contrast to FW trout, SW trout were found to possess a Na⁺–K⁺–Cl^– cotransport process. This finding is regarded as a major adaptation to SW since this cotransport allows an increase of ions and water absorption. Both FW and SW trout were equipped with a Na⁺–H⁺ exchange. In FW, the intestine of the trout had both a Na⁺–Na⁺ exchange and a Na⁺ conductance which may be responsible for enterocyte Na⁺ uptake along the potential gradient.     

Nitrogen Dynamics in the Steeply Stratified,Temperate Lake Verevi,Estonia

The dynamics of different nitrogen compounds and nitrification in diverse habitats of a stratified Lake Verevi (Estonia) was investigated in 2000–2001. Also planktonic N₂-fixation (N₂fix) was measured in August of the observed years. The nitrogen that accumulated in the hypolimnion was trapped in the non-mixed layer during most of the vegetation period causing a concentration of an order of magnitude higher than in the epilimnion. The ammonium level remained low in the epilimnion (maximum 577 mgN m⁻³, average 115 mgN m⁻³) in spite of high concentrations in the hypolimnion (maximum 12223 mgN m⁻³, average 4807 mgN m⁻³). The concentrations of NO₂⁻ and NO₃⁻ remained on a low level both in the epilimnion (average 0.94 and 9.09 mgN m⁻³, respectively) and hypolimnion (average 0.47 and 5.05 mgN m⁻³, respectively). N₂fix and nitrification ranged from 0.30 to 2.80 mgN m⁻³ day⁻¹ and 6.0 to 107 mgN m⁻³ day⁻¹, respectively; the most intensive processes occurred in 07.08.00 at depths of 2 and 5 m, accordingly. The role of N₂fix in the total nitrogen budget of Lake Verevi (in August 2000 and 2001) was negligible while episodically in the nitrogen-depleted epilimnion the N₂fix could substantially contribute to the pool of mineral nitrogen. Nitrification was unable to influence nitrogen dynamics in the epilimnion while some temporary coupling with ammonium dynamics in the hypolimnion was documented.     

Effect of Rural Sewage Irrigation Regime on Water-Nitrogen Utilization and Crop Growth of Paddy Rice in Southern China

Reclaimed water irrigation has become an effective mean to alleviate the contradiction between water availability and its consumption worldwide. In this study, three types of irrigation water sources (rural sewage’s primary treated water R1 and secondary treated water R2, and river water R3) meeting the requirements of water quality for farmland irrigation were selected, and three types of irrigation water levels (low water level W1 of 0–80 mm, medium water level W2 of 0–100 mm, and high water level W3 of 0–150 mm) were adopted to carry out research on the influence mechanismS of different irrigation water sources and water levels on water and nitrogen use and crop growth in paddy field. The water quantity indicators (irrigation times and irrigation volume), soil ammonium nitrogen (NH₄⁺-N) and nitrate nitrogen (NO₃⁻-N), rice yield indicators (thousand-grain weight, the number of grains per spike, and the number of effective spikes), and quality indicators (the amount of protein, amylose, vitamin C, nitrate and nitrite content) of rice were measured. The results showed that, the average irrigation volume under W3 was 2.4 and 1.9 times of that under W1 and W2, respectively. Compared with R3, the peak consumption of rice was lagged behind under R1 and R2, and the nitrogen form in 0–40 cm soil layers under rural sewage irrigation was mainly NH₄⁺-N. The changes of NO₃⁻-N and NH₄⁺-N in the 0–40 cm soil layer showed the trend of declining and then increasing. The water level control only had a significant effect on the change of NO₃⁻-N in the 60–80 cm soil layer. Both irrigation water use efficiency and crop water use efficiency were gradually reduced with the increase of field water level control. The nitrogen utilization efficiency under rural sewage irrigation was significantly higher than that under R3. Compared with the R3, rural sewage irrigation could significantly increase the yield of rice, and as the field water level rose, the effect of yield promotion was more obvious. It was noteworthy that the grain of rice under R1 monitored the low nitrate and nitrite content, but no nitrate and nitrite was discovered under R2 and R3. Therefore, reasonable rural sewage irrigation (R2) and medium water level (W2) were beneficial to improve nitrogen utilization efficiency, crop yield and crop quality promotion.

     

In situ measurement of fine root water absorption in three temperate tree species—Temporal variability and control by soil and atmospheric factors

Miniature heat balance-sap flow gauges were used to measure water flows in small-diameter roots (3–4 mm) in the undisturbed soil of a mature beech–oak–spruce mixed stand. By relating sap flow to the surface area of all branch fine roots distal to the gauge, we were able to calculate real time water uptake rates per root surface area (J_s) for individual fine root systems of 0.5–1.0 m in length. Study aims were (i) to quantify root water uptake of mature trees under field conditions with respect to average rates, and diurnal and seasonal changes of J_s, and (ii) to investigate the relationship between uptake and soil moisture θ, atmospheric saturation deficit D, and radiation I. On most days, water uptake followed the diurnal course of D with a mid-day peak and low night flow. Neighbouring roots of the same species differed up to 10-fold in their daily totals of J_s (<100–2000 g m⁻² d⁻¹) indicating a large spatial heterogeneity in uptake. Beech, oak and spruce roots revealed different seasonal patterns of water uptake although they were extracting water from the same soil volume. Multiple regression analyses on the influence of D, I and θ on root water uptake showed that D was the single most influential environmental factor in beech and oak (variable selection in 77% and 79% of the investigated roots), whereas D was less important in spruce roots (50% variable selection). A comparison of root water uptake with synchronous leaf transpiration (porometer data) indicated that average water fluxes per surface area in the beech and oak trees were about 2.5 and 5.5 times smaller on the uptake side (roots) than on the loss side (leaves) given that all branch roots <2 mm were equally participating in uptake. Beech fine roots showed maximal uptake rates on mid-summer days in the range of 48–205 g m⁻² h⁻¹ (i.e. 0.7–3.2 mmol m⁻² s⁻¹), oak of 12–160 g m⁻² h⁻¹ (0.2–2.5 mmol m⁻² s⁻¹). Maximal transpiration rates ranged from 3 to 5 and from 5 to 6 mmol m⁻² s⁻¹ for sun canopy leaves of beech and oak, respectively. We conclude that instantaneous rates of root water uptake in beech, oak and spruce trees are above all controlled by atmospheric factors. The effects of different root conductivities, soil moisture, and soil hydraulic properties become increasingly important if time spans longer than a week are considered.     

Comparison of the physical properties of real and artificial salmonid eggs and of their performance when drifting in an experimental stream channel

The paper compares three batches of brown trout (Salmo trutta Linnaeus) eggs, two batches of Atlantic salmon (Salmo salar Linnaeus) eggs and the artificial trout eggs described by Ottaway (1981), in terms of their measurable physical characteristics. Attention was given to the effects of temperature and of the stage of development of the real eggs. Comparisons were also made between brown trout eggs and artificial eggs of the rate of fall in a water column, of the value and speed of attainment of terminal velocity when falling through water and of the pattern of settlement within an experimental stream channel.The main measurable characteristics of brown trout eggs (after water hardening) and of artificial eggs (values in parentheses) were: diameter 0.52–0.54 mm (0.54), fresh weight 0.078–0.086 g (0.09), density 1.071–1.075 g ml^–1 (1.066), and volume 0.073–0.081 ml (0.085). There was no evidence of any major change in these values as egg development proceeded. Salmon eggs had a similar density to trout and artificial eggs but had 30 to 70% greater weight and volume. The water-hardening of eggs caused their dry matter content to fall from 37–39% to 31–34% and there were corresponding increases in volume and fresh weight.The rate of fall of trout eggs and artificial eggs through water when timed from rest at the water surface to a depth of 125 cm was similar for both types of egg at 10 °C. This rate rose with increasing temperature at the same low rate of c. 0.02 cm s^–1 °C^–1 for both types of egg. Both types of egg had a terminal velocity of c. 8.8 cm s^–1 and both achieved terminal velocity in less than 1.5 s after release.The patterns of settlement of the two types of eggs in an experimental channel were similar.     

Quantitation of entacapone glucuronide in rat plasma by on-line coupled restricted access media column and liquid chromatography–tandem mass spectrometry

A column-switching liquid chromatography–electrospray ionization-tandem mass spectrometric (LC–ESI-MS–MS) method was developed for the direct analysis of entacapone glucuronide in plasma. The plasma samples (5 μl) were injected onto a C₁₈-alkyl-diol silica (ADS) column and the matrix compounds were washed to waste with a mixture of 20 mM ammonium acetate solution at pH 4.0–acetonitrile (97:3). The retained analyte fraction containing (E)- and (Z)-isomers of glucuronides of entacapone and tolcapone glucuronide (internal standard) was backflushed to the analytical C₁₈ column, with a mixture of 20 mM ammonium acetate–acetonitrile (85:15) for the final separation at pH 7.0. The eluate was directed to the mass spectrometer after splitting (1:100). The mass spectrometer was operated in the negative ion mode and the deprotonated molecules [M−H]⁻ were chosen as precursor ions for the analytes and internal standard. Collisionally induced dissociation of [M−H]⁻ in MS–MS resulted in loss of the neutral glucuronide moiety and in the appearance of intensive negatively charged aglycones [M−H−Glu]⁻, which were chosen as the product ions for single reaction monitoring. Quantitative studies showed a wide dynamic range (0.0025–100 μg/ml) with correlation coefficients better than 0.995. The method was repeatable within-day (relative standard deviation, RSD<7%) and between-day (RSD<14%) and the recovery (78–103%) was better than with the traditional, laborious pretreatment method. The use of tandem mass spectrometry permitted low limits of detection (1 ng/ml of entacapone glucuronide). The method was applied for the quantitation of (E)- and (Z)-isomers of entacapone glucuronide in plasma of rats used in absorption studies.     

Impacts of mussel (<Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis>) farming on oxygen consumption and nutrient recycling in a eutrophic coastal lagoon

Fluxes of oxygen, nitrogen and phosphorus were determined in two areas of the Sacca di Goro lagoon, at a site influenced by the farming of the mussel Mytilus galloprovincialis and a control site. Mussel farming induced intense biodeposition of organic matter to the underlying sediments, which stimulated sediment oxygen demand, and inorganic nitrogen and phosphorus regeneration rates compared to the nearby control station. Overall benthic fluxes (–11.4 ± 6.5 mmol O₂ m⁻² h⁻¹; 1.59 ± 0.47 mmol NH₄⁺ m⁻² h⁻¹ and 94 ± 42 μmol PO₄³⁻ m⁻² h⁻¹) at the mussel farm are amongst the highest ever recorded for an aquaculture impacted area and question the belief that farming of filter-feeding bivalves has inherently lower impacts than finfish farming. In situ incubations of intact mussel ropes demonstrated that the mussel rope community was an enormous sink for oxygen and particulate organic matter, and an equally large source of dissolved inorganic nitrogen and phosphate to the water column. Overall, a one meter square area of␣mussel farm (mussel ropes and underlying sediment) was estimated to have an oxygen demand of 46.8 mmol m² h⁻¹ and to regenerate inorganic nitrogen and phosphorus at rates of 8.5 and 0.3 mmol m² h⁻¹, with the mussel ropes accounting for between 70 and more than 90% of the overall oxygen and nutrient fluxes. Even taking into account that within the farmed area of the Sacca di Goro lagoon, there are 15–20 m⁻² of open water for each one covered with mussel ropes, the mussel ropes would account for a large and often dominant part of overall oxygen and nutrient fluxes. These results demonstrate that it is essential to take into account the activity of the cultivated organisms and their epiphytic community when assessing the impacts of shellfish farming. Overall, whilst grazing by the mussel rope community could act as a top-down control on the phytoplankton, most of the ingested organic matter is rapidly recycled to the water column as inorganic nutrients, which would be expected to stimulate phytoplankton growth. Consequently, the net effect of the mussel farming on phytoplankton dynamics, may be to increase phytoplankton turnover and overall production, rather than to limit phytoplankton biomass.     

The recovery and hematological rehabilitation of chlorine stressed adult rainbow trout (Salmo gairdneri)

Synopsis The ability of adult rainbow trout (Salmo gairdneri) to recover from acute chlorine exposure was tested. Six separate tests were carried out in duplicate. Blood was collected from each duplicate group of fish at the beginning of the test, at the appearance of the chlorine-distress symptoms, and at 24 and 48 h after exposures. The total residual chlorine (TRC) concentrations used in these six tests were 1.67, 3.50, 1.10, 1.25, 1.02 and 0.0 mg 1^–1, at water temperatures of 13.5, 14.6, 17.2, 22.6, 23.0 and 26.0° C, respectively. The cumulative fish mortality for these six tests at 48 h after exposure were 11.5, 100, 0.0, 67.1, 36.1 and 100 percent. Blood of chlorine stressed fish exhibited signs of hemoconcentration and hemolysis. All blood parameters with the exception of hemolysis returned to control levels within 24 hours after exporsure. High TRC concentrations in the range of 3.5 mg 1^–1 at 14.0° C, and TRC of 1.25 and 1.02 mg 1^–1 TRC at 23° C, significantly diminished the rainbow trout recovery.     

Biofiltering efficiency in removal of dissolved nutrients by three species of estuarine macroalgae cultivated with sea bass (Dicentrarchus labrax) waste waters 2. Ammonium

Three estuarine macroalgae (Ulva rotundata,Enteromorpha intestinalis, Gracilariagracilis) of economic potential were cultivated in the laboratory toassess their biofiltering capacities for ammonium in waste effluents from a seabass (Dicentrarchus labrax) cultivation tank. The studywasdeveloped to investigate the functioning of N nutrition of the three species.Atlow water flow (< 2 volumes d^–1) the three species strippedefficiently the ammonium dissolved in the waste water from the fish tank, withaminimum biofiltering efficiency estimate of 61% in unstarved cultures ofG. gracilis at a water flow of 2 volumesd^–1. Maximum velocity for ammonium uptake (89.0 molNH₄ ⁺ g^–1 dry wth^–1) was found in U. rotundata,whereas G. gracilis showed the highest affinity for thisnutrient. The net ammonium uptake rate was significantly affected by the waterflow, being greatest at the highest flow assayed (2 volumesd^–1). Variations of tissue N and C:N ratios during aflow-through experiment suggested that N was not limiting macroalgal growth.However, when ammonium was supplied at a flow rate of 0.5 volumesd^–1, specially in a three-stage design, the marked reductionintissue N and the biomass C:N:P ratios suggested a more general nutrientdeficiency. A significant correlation was found between growth rates and the Nbiomass gained in the cultures. The three-stage design under low water flow(0.5volumes d^–1) showed that the highest ammonium uptake rates (upto 80.9 mol NH₄ ⁺ g^–1 dry wtd^–1 in U. rotundata) were found inthe first stage, with decreasing rates in the following ones. As a result, lowincrements or even losses of total N biomass in these stages were found,suggesting that ammonium was excreted from the algae. We conclude that thesespecies present a potential ability to biofilter the ammonium dissolved inwastewater from a D. labrax cultivation tank, suggesting thatscaling up the biofiltration designs, future practises using these macroalgaemay be implemented in the local fish farms, resulting in both environmental andeconomical advantages.     

Growth, uptake, and assimilation of ammonium, nitrate, and urea, by three strains of Karenia brevis grown under low light

Observations of near-bottom populations of Karenia brevis suggest that these cells may derive nutrients from the sediment–water interface. Cells undergoing a metabolic-mediated migration may be in close proximity to enhanced concentrations of nutrients associated with the sediment during at least a fraction of their diel cycle. In this study, the growth, uptake and assimilation rates of ammonium, nitrate, and urea by K. brevis were examined on a diel basis to better understand the potential role of these nutrients in the near-bottom ecology of this species. Three strains of K. brevis, C6, C3, and CCMP 2229, were grown under 12:12 light dark cycle under 30 μmol photons m⁻² s⁻¹ delivered to the surface plain of batch cultures. Nitrogen uptake was evaluated using ¹⁵N tracer techniques and trichloroacetic acid extraction was used to evaluate the quantity of nitrogen (N) assimilated into cell protein. Growth rates ranged from a low of 0.12 divisions day⁻¹ for C6 and C3 grown on nitrate to a high of 0.18 divisions day⁻¹ for C3 grown on urea. Diurnal maximum uptake rates, ρ_max, varied from 0.41 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on nitrate, to 1.29 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on urea. Average nocturnal uptake rates were 29% of diurnal rates for nitrate, 103% of diurnal uptake rates for ammonium and 56% of diurnal uptake rates for urea. Uptake kinetic parameters varied between substrates, between strains and between day and night measurements. Highest maximum uptake rates were found for urea for strains CCMP2229 and C3 and for ammonium for strain C6. Rates of asmilation into protein also varied day and night, but overall were highest for urea. The comparison of maximal uptake rates as well as assimilation efficiencies indicate that ammonium and urea are utilized (taken up and assimilated) more than twice was fast as nitrate on a diel basis.     

Characterization, dynamics, and ecological impacts of harmful Cochlodinium polykrikoides blooms on eastern Long Island, NY, USA

We report on the emergence of Cochlodinium polykrikoides blooms in the Peconic Estuary and Shinnecock Bay, NY, USA, during 2002–2006. Blooms occurred during late summer when temperatures and salinities ranged from 20 to 25 °C and 22 to 30 ppt, respectively. Bloom patches achieved cell densities exceeding 10⁵ ml⁻¹ and chlorophyll a levels exceeding 100 μg l⁻¹, while background bloom densities were typically 10³–10⁴ cells ml⁻¹. Light, scanning electron and ultrathin-section transmission electron microscopy suggested that cells isolated from blooms displayed characteristics of C. polykrikoides and provide the first clear documentation of the fine structure for this species. Sequencing of a hypervariable region of the large subunit rDNA confirmed this finding, displaying 100% similarity to other North American C. polykrikoides strains, but a lower similarity to strains from Southeast Asia (88–90%). Bioassay experiments demonstrated that 24 h exposure to bloom waters (>5 × 10⁴ cells ml⁻¹) killed 100% of multiple fish species (1-week-old Cyprinodon variegates, adult Fundulus majalis, adult Menidia menidia) and 80% of adult Fundulus heteroclitus. Microscopic evaluation of the gills of moribund fish revealed epithelial proliferation with focal areas of fusion of gill lamellae, suggesting impairment of gill function (e.g. respiration, nitrogen excretion, ion balance). Lower fish mortality was observed at intermediate C. polykrikoides densities (10³–10⁴ cells ml⁻¹), while fish survived for 48 h at cell densities below 1 × 10³ cells ml⁻¹. The inability of frozen and thawed-, or filtered (0.2 μm)-bloom water to cause fish mortality suggested that the thick polysaccharide layer associated with cell membranes and/or a toxin principle within this layer may be responsible for fish mortality. Juvenile bay scallops (Argopecten irradians) and American oysters (Crassostrea virginica) experienced elevated mortality compared to control treatments during a 9-day exposure to bloom water (5 × 10⁴ cells ml⁻¹). Surviving scallops exposed to bloom water also experienced significantly reduced growth rates. Moribund shellfish displayed hyperplasia, hemorrhaging, squamation, and apoptosis in gill and digestive tissues with gill inflammation specifically associated with areas containing C. polykrikoides cells. In summary, our results indicate C. polykrikoides blooms have become annual events on eastern Long Island and that bloom waters are capable of causing rapid mortality in multiple species of finfish and shellfish.     

Water conservation and protein metabolism in northern elephant seal pups during the postweaning fast

Urine production and N output were monitored in northern elephant seal (Mirounga angustirostris) pups progressing through 10 weeks of a natural postweaning fast. Urine output declind by 84% (to 69±12 ml·day^–1) at 10 weeks (P<0.05). Glomerular filtration rate at 10 weeks was 51% of the 67±3 ml serum·min^–1 observed during week 1 (P<0.05). Urine N excretion fell by 69% to 1.2±0.17 g·day^–1, while urinary concentration increased (P<0.05). Serum urea declined from an initial 11 mmol·1^–1 to 5–7 mmol·1^–1 by 5 weeks. The fall in urinary N loss (and thus amino acid oxidation) was concomitant with depressed metabolic rate. Therefore, protein contributed little toward meeting energy demands (i.e., <4% of average metabolic rate) throughout fasting. These data indicate that fasting pups improve water conservation and minimize protein catabolism during prolonged natural fasts without an exogenous source of water.Abbreviations AA amino acid(s) - AMR average metabolic rate - ANOVA one-way analysis of variance - BMR basal metabolic rate - BUN blood urea nitrogen - EP end product - EWL evaporative water loss - [Gr]_s serum creatinine concentration - GFR glomerular filtration rate - LBM lean body mass - LML Long Marine Laboratory - MR metabolic rate - NEFA non-esterified fatty acids - RMR resting metabolic rate - TCA tricarboxylic acid - U:C ulinary urea: creatinine concentration ratio     

Emission of N2O, N2, CH4, and CO2 from constructed wetlands for wastewater treatment and from riparian buffer zones

We measured nitrous oxide (N₂O), dinitrogen (N₂), methane (CH₄), and carbon dioxide (CO₂) fluxes in horizontal and vertical flow constructed wetlands (CW) and in a riparian alder stand in southern Estonia using the closed chamber method in the period from October 2001 to November 2003. The replicates’ average values of N₂O, N₂, CH₄ and CO₂ fluxes from the riparian gray alder stand varied from −0.4 to 58 μg N₂O-N m⁻² h⁻¹, 0.02–17.4 mg N₂-N m⁻² h⁻¹, 0.1–265 μg CH₄-C m⁻² h⁻¹ and 55–61 mg CO₂-C m⁻² h⁻¹, respectively. In horizontal subsurface flow (HSSF) beds of CWs, the average N₂ emission varied from 0.17 to 130 and from 0.33 to 119 mg N₂-N m⁻² h⁻¹ in the vertical subsurface flow (VSSF) beds. The average N₂O-N emission from the microsites above the inflow pipes of the HSSF CWs was 6.4–31 μg N₂O-N m⁻² h⁻¹, whereas the outflow microsites emitted 2.4–8 μg N₂O-N m⁻² h⁻¹. In VSSF beds, the same value was 35.6–44.7 μg N₂O-N m⁻² h⁻¹. The average CH₄ emission from the inflow and outflow microsites in the HSSF CWs differed significantly, ranging from 640 to 9715 and from 30 to 770 μg CH₄-C m⁻² h⁻¹, respectively. The average CO₂ emission was somewhat higher in VSSF beds (140–291 mg CO₂-C m⁻² h⁻¹) and at the inflow microsites of HSSF beds (61–140 mg CO₂-C m⁻² h⁻¹). The global warming potential (GWP) from N₂O and CH₄ was comparatively high in both types of CWs (4.8 ± 9.8 and 6.8 ± 16.2 t CO₂ eq ha⁻¹ a⁻¹ in the HSSF CW 6.5 ± 13.0 and 5.3 ± 24.7 t CO₂ eq ha⁻¹ a⁻¹ in the hybrid CW, respectively). The GWP of the riparian alder forest from both N₂O and CH₄ was relatively low (0.4 ± 1.0 and 0.1 ± 0.30 t CO₂ eq ha⁻¹ a⁻¹, respectively), whereas the CO₂-C flux was remarkable (3.5 ± 3.7 t ha⁻¹ a⁻¹). The global influence of CWs is not significant. Even if all global domestic wastewater were treated by wetlands, their share of the trace gas emission budget would be less than 1%.     

Ion and acid–base regulation in the freshwater mummichog (Fundulus heteroclitus): a departure from the standard model for freshwater teleosts

Ion and acid–base balance were examined in the freshwater-adapted mummichog (Fundulus heteroclitus) using a series of treatments designed to perturb the coupling mechanisms. Unidirectional Cl⁻ uptake (J^Cl_in) was extremely low whereas J^Na_in was substantial (three- to sixfold higher); comparable differences occurred in unidirectional efflux rates (J^Cl_out, J^Na_out). J^Cl_in was refractory to all treatments, suggesting that Cl⁻/base exchange was unimportant or absent. Indeed, no base excretion or modulation of ion fluxes occurred for acid–base balance for up to 8 h after NaHCO₃⁻ loading (injections of 1000 or 3000 nequiv.·g⁻¹). Acute environmental low pH (4.5) and amiloride (10⁻⁴ M) treatments caused concurrent inhibition of J^Na_in and net H⁺ excretion (J^H+_net), indicating the presence of Na⁺/H⁺ exchange. J^Na_in was elevated and J^H+_net restored during recovery from both treatments, but this exchange did not appear to be dynamically adjusted for acid–base homeostasis. High external ammonia exposure (1 mmol·l⁻¹) initially blocked ammonia excretion (J^Amm_net) but had no effect on J^Na_in, whereas high pH (9.4) reduced both J^Amm_net and J^Na_in. Inhibition of J^Na_in by the low pH and amiloride treatments had no effect on J^Amm_net. These results indicate that ammonia excretion is entirely diffusive and independent of both Na⁺uptake and the protons that are transported via the Na⁺/H⁺ coupling. In addition, ureagenesis served as a compensatory mechanism during high external ammonia exposure, as a marked elevation in urea excretion partially replaced the inhibited J^Amm_net. In all treatments, changes in the Na⁺–Cl⁻ net flux differential were consistent with changes in J^H+_net measured by traditional water titration techniques, indicating that the former can be used as an estimate of the acid–base status of the fish. Overall, the results demonstrate that the freshwater-adapted F. heteroclitus does not conform to the ion/acid–base relationships described in the standard model based on commonly studied species such as trout, goldfish, and catfish.     

Ground water flow paths in relation to nitrogen chemistry in the near-stream zone

Interactions between ground water flow paths and water chemistry were studied in the riparian zone of a small headwater catchment near Toronto, Ontario. Significant variations in oxygen — 18 and chloride indicated the presence of distinct sources of water in the ground water flow system entering the near-stream zone. Shallow ground water at the upland perimeter of the riparian zone had nitrate-N, chloride and dissolved oxygen concentrations which ranged between 100–180 µg L^–1, 1.2–1.8 mg L^–1 and 4.6–9.1 mg L^–1 respectively. Concentrations of nitrate — N in deep ground water flowing upward beneath the riparian wetland were < 10 µg L^–1, whereas chloride and dissolved oxygen ranged between 0.6–0.9 mg L^–1 and 0.4–2.2 mg L^–1 respectively. Ammonium — N concentrations (20–60 µg L^–1) were similar in shallow and deep ground water. Ground water was transported through the wetland to the stream by three hydrologic pathways. 1) Shallow ground water emerged as springs near the base of the hillslope producing surface rivulets which crossed the riparian zone to the stream. 2) Deep ground water flowed upward through organic soils and entered the rivulets within the wetland. 3) Deep ground water reached the stream as bed and bank seepage. Springs were higher in nitrate and chloride than rivulets entering the stream, whereas bank seeps had lower concentrations of nitrate and chloride and considerably higher ammonium concentrations than the rivulets. These contrasts in nitrate and chloride concentrations were related to initial differences in the ion chemistry of shallow and deep ground water rather than to element transformations within the riparian wetland. Differences in ammonium concentration between seeps and rivulets were caused by immobilization of ammonium in the substrates of aerobic rivulets, whereas little ammonium depletion probably occurred in deep ground water flowing upward through reduced subsurface organic soils around the stream perimeter.     

Growth characters and photosynthetic capacity of <Emphasis Type="BoldItalic">Gracilaria lemaneiformis</Emphasis> as a biofilter in a shellfish farming area in Sanggou Bay,China

Experiments on growth characters and ecological functions of the macroalgae Gracilaria lemaneiformis, collected from south China, were conducted in polyculture areas of kelp and filter-feeding bivalve in Sanggou Bay in Weihai City, Shandong, in north China from May 2002 to May 2003. The results of 116 days cultivation showed that the average wet weight of alga increased 89 times from 0.1 to 8.9 kg rope⁻¹, with an average specific growth rate (based on wet weight) of 3.95% per day. The most favorable water layer for its growth was 1.0–1.8 m below the surface in July and August, with an average specific growth rate of 8.2% per day in 30-day experiments. Photosynthetic activity changed seasonally, with an average of 7.3 mg O₂ g dw⁻¹ h⁻¹. The maximum rate (14.4 mg O₂ g dw⁻¹ h⁻¹) was recorded in July, or 19.3 mg CO₂ g dw⁻¹ h⁻¹, while the minimum (0.40 mg CO₂ g dw⁻¹ h⁻¹) was in April. This study indicated that the culture of G. lemaneiformis is an effective way to improve water quality where scallops are cultivated intensively.     

Cardio-respiratory effects of chloramine-T exposure in rainbow trout

In order to establish whether the blood gas respiratory disturbances noted with exposure to chloramine-T are due to differences in the rates of uptake of O₂ and excretion of CO₂ or gill blood flow, adult rainbow trout (Oncorhynchus mykiss) were fitted with dorsal aorta and bulbus arteriosus catheters to facilitate blood pressure recordings, an ultrasonic blood flow probe and opercular impedance electrodes. Fish received either a 45-min static exposure to 9 mg l⁻¹ chloramine-T or tap water (control) and continuous recordings of blood pressure, and ventilation frequency and amplitude were made. Pre- and post-exposure arterial and venous blood samples were taken and analyzed for O₂ and CO₂ content, hemoglobin concentration and hematocrit. Chloramine-T exposure had no effect on any of the continuously recorded parameters. However, individual measurements (made immediately prior to and following exposure) of cardiac output and O₂ uptake rates increased significantly following exposure to chloramine-T compared to before exposure. CO₂ excretion rates were unaffected by chloramine-T exposure. Calculation of the perfusion convection requirement showed a significant increase for CO₂ but not for O₂. It was concluded that increases in O₂ uptake resulted from increased cardiac output but that CO₂ excretion, a diffusion-limited process, was not increased due to additional diffusive limitations caused by the irritant effect of chloramine-T.