Surface nutrients,chlorophyll a and phaeopigment in some Scottish sea lochs

The distribution of several surface properties, both physical and chemical, of three Scottish sea lochs were studied during the spring phytoplankton bloom.Concentrations of nitrate, phosphate, and silicate have been determined and related to fluctuations in the freshwater input and uptake by the phytoplankton population. The ratios nitrate: phosphate and silicate: phosphate (on atom bases) in Loch Creran and Loch Linnhe were similar, but there were certain marked differences from those in Loch Etive.Nitrate, rather than phosphate, was a critical factor in the productivity of Loch Creran and Loch Linnhe. In Loch Etive the limitation of these nutrients was directly affected by the amount of fresh-water entering into the Loch.Patchiness of chlorophyll a and phaeopigments in Loch Creran and Loch Etive reflected those factors (e.g., freshwater/sea-water inflow, turbulence produced by the loch topography etc.) which affect the surface circulation in the two lochs.The delay in the development of the phytoplankton bloom in Loch Linnhe may be attributed to a poor stability of the surface layers due to low run-off.     

Chemical investigations of Loch Etive,Scotland. II. Dissolved organic compounds

Variations in the levels of dissolved organic compounds in Loch Etive were studied during 1971. The amount of dissolved organic nitrogen, dissolved organic phosphorus, and Gelbstoff were markedly affected by the freshwater run-off. The surface (1 m) concentrations of the dissolved organic nitrogen and dissolved organic phosphorus throughout the year ranged from 3.0–9.0 μg-at. NO₃-N/l and 0.02-0.45 μg-at. PO₄-P/1, respectively. The surface levels of Gelbstoff during the year as measured by the optical density of the water at 320 nm and 400 nm fluctuated between 0.08-0.490 and 0.020-0.140, respectively. In the bottom layers of the Lower Basin (40–50 m) the optical density of the water at 320 nm and 400 nm throughout the year ranged between 0.060-0.150 and 0.010-0.047, in the Deep Basin (115–140 m) the values were between 0.086-0.130 and 0.018-0.037 and at the Head of the Loch, (25–35 m) they fluctuated between 0.080-0.229 and 0.017-0.052, respectively. The vertical concentrations of the dissolved organic nitrogen, dissolved organic phosphorus and Gelbstoff decreased irregularly from the surface downward. The concentrations of the dissolved organic nitrogen at the surface of the Lower Basin during the year were similar to those found at 40–50 m. In the Deep Basin and at the Head of the Loch the surface values were higher than those observed at 115–140 m and 25–35 m, respectively. The concentrations of the dissolved organic phosphorus at the surface of the Lower Basin were close to those found at the bottom (40–50 m). In the Deep Basin the surface values were higher than those at 115–140 m and at the Head of the Loch the surface concentrations were variably higher or lower than the bottom layers (25–35 m). The ratio of the optical density at 320 nm and 400 nm in the photic zone (1 m, 4–5 m, and 10 m) during the year ranged between 3.2–4.0 with the lowest ratios in winter and autumn. A similar pattern was observed in the water column of the Lower Basin and at the Head of the Loch. In the deeper strata of the Deep Basin, ratios were usually between 4.0–5.3 for most of the year. A comparison between the optical densities of the surface water during winter and autumn and that at 115–140 m (Deep Basin), suggests that the fraction more liable to precipitate is found in the visible region of the spectrum (400 nm).     

Azote et phosphore chez un crustacé macroplanctonique, Meganyctiphanes norvegica (M. Sars) (Euphausiacea): Excrétion minérale et constitution

Ammonia and phosphate excretion by the Mediterranean euphausiid Meganyctiphanes norvegica (M. Sars) (mean individual dry weight, 60 mg) has been measured for one year. Experiments were conducted at 13°C on single, freshly caught animals maintained in unfiltered sea water. Possible influence of these experimental procedures upon values obtained are discussed. Phytoplankton re-uptake and bacterial activity proved insignificant, because of the short duration of the experiments (4–38 h). NH₄ and PO₄ excretion rates are higher shortly after collection and then decrease, reaching a steady level after 8 h for NH₄, but continuously decreasing during the 38 h of the longer experiment in the c case of PO₄. It is considered that earlier higher values are likely to be more representative of in situ rates despite possible ‘stress’ effects, because they are close to those of moderately fed animals kept in captivity; more stable values observed after 8–12 h are close to those of starved animals. Nevertheless, lower stabilized values are best used when investigating seasonal variations. Excretion rates are low (0.07 to 0.11 μg-at.NH₄-N.mg^?1.day^?1, and 0.009 to 0.010 μ-at.PO₄-P mg^?1.day^?1) in summer, autumn and early winter. They rise sharply from January–February (0.12 μg-at.NH₄-N.mg^?1.day^?1 and 0.015 μg-at.PO₄-P.mg^?.day^?1) to peak spring values (0.25 μg-at.NH₄-N.mg^?1.day^?1 and 0.026 μg-at. PO₄-P.mg^?1.day^?1). The significance of inorganic excretion with regard to total (inorganic + organic) excretion is discussed. The nitrogen and phosphorus content of the animals were simultaneously measured and amount, respectively, to 9.5 and 0.8% of body dry weight (mean yearly value). Based on inorganic excretion only, the mean values of turnover are 66 days for nitrogen (61–92 days from May to February, 28–32 days in March–April) and 16 days for phosphorus (12–22 days with limited seasonal variation). Mean N/P ratio by atoms for excretion amounts to 9.1 after 8–12 h (it increases afterwards due to continual decrease in PO₄ excretion) but there are significant seasonal variations. The mean N/P ratio by atoms for the animals is 27.4; it is lower (22.4) between March and August, and higher (29.2) from September to February. With a view to investigating the change in the N/P ratio in the chain prey → predator → excretion, which allows calculation of growth efficiency factor (k₂) and hence secondary production, the nitrogen and phosphorus of the stomach contents of the animals were measured. Due to unknown bias (possibly a terrestrial origin of food particles, or loss of phosphorus during conservation of samples), the results were disappointing (N/P by atoms = 69, instead of a value necessarily ranging between 9.1 and 27.4), and did not allow calculation of k₂.     

Chemical investigations of Loch Etive,Scotland. III. Participate organic carbon and particulate organic nitrogen

The concentration of the suspended matter in Loch Etive varies with river discharge, biological production, and water movement. The contribution from phytoplankton production shows a seasonal trend, with the highest values in the spring and the lowest in winter. The carbon: chlorophyll a and nitrogen: chlorophyll a ratios in the photic zone (1,4–5, and 10 m) during the period of high production ranged from 54.0 to 400.0 and 6.0 to 38.0, respectively, the autumn bloom having higher ratios than the spring bloom. The increase of these ratios in the landward direction indicates the influence of terrestrial material. The C/N ratios in the surface (1 m) of the loch throughout the year fluctuated between 6.5–20.0, being the highest in winter. The C/N ratios in the bottom layers of the Lower Basin (40–50 m), Deep Basin (115–140 m) and the Head of the Loch (25–35 m) during the year ranged between 2.25–14.0, 5.0–38.0, and 3.0–28.5, respectively. The lowest ratios were mainly found in early autumn and associated with high levels of dissolved organic nitrogen and amonium. The accumulation of suspended matter in the deeper layers of the Deep Basin was caused by the input of organic detritus from the Lower Basin and eroded sediment from the basin slope. The lateral distribution of the C/N ratios may be related to the hydrographical features of the loch.     

AMMONIUN AND NITRATE UPTAKE BY THE MARINE MACROPHYTES HYPNEA MUSVUFORMIS (RHODOPHYTA) AND MACROCYSTIS PYRIFERA (PHAEOPHYTA)1, 2

NH₄⁺ and NO₃^? uptake were measured by continuous sampling with an autoanalyzer. For Hypnea musciformis (Wulfen) Lamouroux, NO₃^?up take followed saturable kinetics (K₂=4.9 μg-at N t^?1, V_max= 2.85 μg- at N, g(wet)^?1. h^?1. The ammonium uptake data fit a trucatd hyperbola, i.e., saturation was not reach at the concentrations used. NO₃^? uptake was reduced one-half in the presence of NH₄⁺, but presence of NO₃^? had no effect on NH₄⁺ uptake. Darkness reduced both NO₃^? and NH₄⁺ uptake by one-third to one-half. For Macrocystis pyrufera (L) C. Agardh, NO₃^? uptake followed saturable kinetices: K₂=13.1 μg-at N. l^?1. V_max=3.05 μg-at N. g(wet)^?1. h^?1.NH₄⁺ uptake showed saturable kinetics at concentration below 22 μg-at N l -1 (K₂=5.3 μg-at N.1–1, V_max= 2.38 μg-at N G (wet)^?1.h^?1: at higher concentration uptake increased lincarly with concentrations. NO₃^?and NH₄⁺ were taken up simulataneously: presence of one form did not affect uptake of the other.     

Three species of Mytilus and their hybrids identified in a Scottish Loch: natives, relicts and invaders?

Three species of the mussel, Mytilus, occur in the North Atlantic region, M. edulis, M. galloprovincialis and M. trossulus, and hybrid zones are present where their distributions overlap. M. edulis is a native species in the UK. M. galloprovincialis originated in the Mediterranean and its distribution extends northwards along the Atlantic seaboard to Scotland. Baltic Sea mussels have a M. trossulus ancestry but are highly introgressed by M. edulis. In recent decades, farming of mussels on long-line rope culture systems has been introduced into Scotland. On farms in Loch Etive, a form of mussel with a fragile shell and a different shape to either M. edulis or M. galloprovincialis has been increasing in frequency over recent years. Samples of fragile shelled, normal strong shelled and intermediate mussel types were sampled from two farms in 2006 and compared with samples of M. edulis, M. galloprovincialis and M. trossulus from other sources where their species identity is well established. Abundance relative to depth, shell strength, condition index and shell morphology were analysed together with 5 allozyme loci and one nuclear DNA genetic marker (Me 15/16). The fragile shelled mussels, and many of those classed as intermediate, were identified as a mixture of M. trossulus and M. trossulus x M. edulis hybrids. This identification was strongly supported by both morphological and genetic data and is the first record of the presence of M. trossulus in UK waters. M. trossulus in Loch Etive are most likely to be a post-glacial relict population restricted to the low salinity area of the Loch that has recently increased in abundance due to commercial mussel growing activity. In addition, individual mussels of all three species and their hybrids were detected amongst Loch Etive mussels. This is the first genetic demonstration of all three species and their hybrids occurring together in one location in the Atlantic region and provides a unique opportunity to study the processes of speciation, divergence, and introgression in the genus Mytilus.     

PHOSPHATE METABOLISM IN BLUE-GREEN ALGAE. II. CHANGES IN PHOSPHATE DISTRIBUTION DURING STARVATION AND THE “POLYPHOSPHATE OVERPLUS” PHENOMENON IN PLECTONEMA BORYANUM

Physiological aspects of phosphate utilization by the blue-green alga Plectonema boryanum were studied. It was found that the external phosphate concentration influenced the distribution of phosphorus-containing compounds in the cell. Culturing the alga in concentrations of 10, 100, and 1000 mg PO₄/l resulted in increases in the level of acid-soluble and acid-insoluble polyphosphates. The values reported for 100 and 1000 mg PO₄/l were the same, indicating that the cells were able to assimilate and utilize only fixed amounts of phosphates. The total phosphorus value for these cells was calculated to be 6.5 μg P per 10⁶ cells. Culturing the alga in 1 mg PO₄/l led to a decrease in phosphate concentration of all cell fractions. Cells grown in the absence of phosphate for 5 days had total cell phosphorus levels of 0.76 μg P per 10⁶ cells. Cells in culture for two months or longer were found to have total cell phosphorus levels of 0.73 μg P per 10⁶ cells. This was determined to be the minimum cell phosphorus level limiting growth. Transfer of cells from either culture condition to a medium containing phosphate led to an “overplus” phenomenon. This overplus phenomenon was characterized by increases in all cellular phosphorus fractions. The most dramatic increase was found in both the acid-soluble and acid-insoluble polyphosphates. These fractions often increased by more than an order of magnitude. The greatest phosphate uptake occurred within 1 hr of transfer of phosphate-starved cells into a medium containing a known amount of phosphate and is essentially complete at 4 hr. The total cell phosphorus levels for uptake never increased beyond 18.9 μg per 10⁶ cells.     

Ammonium formation in cape timiris (Mauritania) upwelling

Ammonium formation was studied in two consecutive years by following a newly upwelled ammonium-free ‘parcel’ of water with a drogue. High concentrations (> 2 μg-at. NH₄-N l^?1) were found at the end of each study. The relative importance of WP-2 net mesozooplankton excretion on the ammonium values is greater before the phytoplanktonic bloom (35–48%) than during it (10–16%). From carbon grazing estimates, it is shown that other herbivore excretion does not take the place of WP-2 mesozooplankton and because of synchronous variations of chlorophyll with dissolved organic nitrogen, and bacterial activity with ammonium, it is suggested that much of the ammonium is produced from phytoplankton decay or nitrogen excretion during the bloom.     

Role of Cations in Accumulation and Release of Phosphate by Acinetobacter Strain 210A

Cells of the strictly aerobic Acinetobacter strain 210A, containing aerobically large amounts of polyphosphate (100 mg of phosphorus per g [dry weight] of biomass), released in the absence of oxygen 1.49 mmol of P_i, 0.77 meq of Mg²⁺, 0.48 meq of K⁺, 0.02 meq of Ca²⁺, and 0.14 meq of NH₄⁺ per g (dry weight) of biomass. The drop in pH during this anaerobic phase was caused by the release of 1.8 protons per PO₄³⁻ molecule. Cells of Acinetobacter strain 132, which do not accumulate polyphosphate aerobically, released only 0.33 mmol of P_i and 0.13 meq of Mg²⁺ per g (dry weight) of biomass but released K⁺ in amounts comparable to those released by strain 210A. Stationary-phase cultures of Acinetobacter strain 210A, in which polyphosphate could not be detected by Neisser staining, aerobically took up phosphate simultaneously with Mg²⁺, the most important counterion in polyphosphate. In the absence of dissolved phosphate in the medium, no Mg²⁺ was taken up. Cells containing polyphosphate granules were able to grow in a Mg-free medium, whereas cells without these granules were not. Mg²⁺ was not essential as a counterion because it could be replaced by Ca²⁺. The presence of small amounts of K⁺ was essential for polyphosphate formation in cells of strain 210A. During continuous cultivation under K⁺ limitation, cells of Acinetobacter strain 210A contained only 14 mg of phosphorus per g (dry weight) of biomass, whereas this element was accumulated in amounts of 59 mg/g under substrate limitation and 41 mg/g under Mg²⁺ limitation. For phosphate uptake in activated sludge, the presence of K⁺ seemed to be crucial.     

Some physical,chemical, and microbiological characteristics of two beaches of anglesey

Observations during 1971 and 1972 of some of the physical, chemical, and microbiological characteristics of contrasting Anglesey beaches, Newborough and Llanddona, are reported. The fine sandy beach at Newborough was observed to be very unstable and topographical changes were recorded. In particular, the movement of a sand wave across the intertidal zone from low water to extinction at the foot of the dune system was observed. The more extensive fine sandy beach at Llanddona had greater stability.Chemically, each beach was variable both spatially and temporally, with ill-defined patterns of concentration changes. Sand from Newborough beach was low in organic carbon (0.07–0.40 mg C/g dry sand) and well aerated, and the soluble inorganic nitrogen in the ground water (up to 30 μg-at. N/l) was dominated by nitrate form (up to 22 μg NO₃-N/l). By contrast, Llanddona sand had a more variable organic carbon content (0.22–2.25 mg C/g dry sand), was wetter, and poorly aerated with consequent sulphide lenses; its dissolved inorganic nitrogen (over 70 μg-at. N/l) was completely dominated by the ammonium form.Microbiologically, the beaches possessed dissimilar bacterial floras, and sediment from Llanddona gave higher bacterial counts than that from Newborough. For both beaches it is shown that estimated bacterial numbers decreased with depth as well as down the intertidal zone.     

Summer metabolism and seasonal changes in biochemical composition of the Antarctic brachiopod Liothyrella uva (Broderip, 1833)

Oxygen consumption and ammonia excretion were measured in Liothyrella uva (Broderip, 1833) held in the laboratory under simulated summer conditions. The mean oxygen consumption of a 50 mg AFDW brachiopod was 0.205 μg-at. O · h ⁻¹ while the ammonia excreted was 0.022 μg-at. NH₃-N · h ⁻¹. The exponents of the regressions relating these parameters to AFDW were not significantly different from each other and the common slope was 0.83. The rise in metabolism over previously published winter levels was 15–20%. The difference between simulated summer and winter conditions was the presence or absence of food. The rise in metabolism was seen in both oxygen consumption and ammonia excretion whilst the oxygen to nitrogen ratio remained virtually unchanged at 9.2 (9.3 in winter), showing that, as in winter conditions, protein was the main metabolic substrate. Oxygen consumption rates were around five times lower than those of temperate brachiopods, even after applying a Q₁₀ temperature correction, and they were two to seven times lower than Antarctic bivalve and gastropod molluscs. An investigation of the biochemical components of field specimens showed strong seasonal variation in protein and carbohydrate, although carbohydrate levels were only as ≈5–10% of protein contents at any one time. There was no significant seasonal variation in lipids, although there was some evidence to suggest that they may be involved, with protein, in the gametogenic cycle. Shell tissues (the caeca) were identified as a site of nutrient storage and may be implicated in short-term storage of material for reproduction as well as the long-term storage of overwintering energy reserves.     

Long term nutrient loads and chlorophyll dynamics in a large temperate Australian lagoon system affected by recurring blooms of cyanobacteria

While rare globally, blooms of the toxic cyanobacteria Nodularia spumigena are a recurring problem in a few estuaries, such as the Baltic Sea and several southern Australian estuaries. Here, we document recurring Nodularia spumigena Mertens blooms in the Gippsland Lakes, S.E. Australia; a temperate lagoon system with episodic, winter-spring dominated catchment inflows. Physico-chemical conditions exerted a strong influence over bloom development, with blooms consistently occurring at surface water salinities between 9 and 20 (average?=?15), inorganic nitrogen concentrations <0.4?μM, and inorganic nitrogen to reactive phosphorus ratios <5. There was a positive correlation between average annual chlorophyll a and total phosphorus (TP) load in years when there was no Nodularia bloom, but this relationship broke down in Nodularia bloom years, even though there was a strong correlation between in-lake TP and chlorophyll a during these years; this highlights the importance of internal sources of phosphorus to bloom development. Large catchment derived nitrate and nitrite (NO_x) inputs following wildfires and floods in 2007, led to high concentrations of NO_x within the surface waters of the Gippsland Lakes through the second half of 2007 and the start of 2008. We hypothesise that these high NO_x concentrations were a key factor leading to an unprecedented Synechococcus sp. bloom that developed in the austral summer of 2007–2008, despite conditions that would otherwise favour a Nodularia bloom.     

Simultaneous quantitative analysis of methyl salicylate,ethyl salicylate and salicylic acid from biological fluids using gas chromatography–mass spectrometry

A gas chromatographic–mass spectrometric (GC–MS) assay was developed for the quantitative analysis of methyl salicylate (MeS), ethyl salicylate (ES) and salicylic acid (SA) from biological fluids. The method was validated from 100-μl rat liver homogenate preparations (5 mg/ml protein) in 70 mM KH₂PO₄ (pH 7.4) buffer and from 100 μl rat plasma. The samples were extracted with chloroform, derivatized with BSTFA and quantitated by GC–MS in the SIM mode. The standard curves ranged from 31 ng/ml to 800 or 1250 ng/ml. Relative standard deviations and bias were less than 11% in plasma and homogenate for all compounds except SA which evidenced greater variability. The assay was used in preliminary experiments to characterize the pharmacokinetics of MeS in rats.     

Growth of the plankton diatom Thalassiosira nordenskioeldii Cleve at low silicate concentrations

The combined effect of low silicate concentration and temperature on the growth of the marine plankton diatom Thalassiosira nordenskioeldii Cleve was investigated by means of batch and semi-continuous cultures. Growth rates were measured in thin cell suspensions (less than 500 cells/ml) to prevent the silicate concentration in the medium from decreasing by more than 25 % during the period of measurement.Half-saturation constants of silicate-limited growth were calculated according to the Michaelis-Menten equation. At 3 °C, the constant was 0.09 μg-at. Si/I and at 10 °C, 0.02μg-at. Si/I. In semicontinuous cultures grown for one month at silicate concentrations of 0.3-0.4μg-at. Si/I, the mean cell division rates were 80–100% of the maximum rates recorded at the respective temperatures.It seems unlikely that decreasing silicate concentrations could influence the course of the spring succession of plankton diatom species in arctic or temperate coastal waters.     

Extracellular phosphomannan as a phosphate reserve in the yeast Kuraishia capsulata

We have found that extracellular phosphomannan is the main phosphate reserve in the yeast Kuraishia capsulata, in contrast to other yeast species effectively absorbing P_i. Under nitrogen starvation, K. capsulata absorbed essentially all P_i from the medium containing 240 mM glucose, 2.5 mM MgSO₄, and 11 mM KH₂PO₄. Inorganic polyphosphate level in the cells was about 14% of the P_i absorbed. Most of the P_i (～60%) was found in the fraction of extracellular phosphomannan that can be used as a carbon and phosphorus source by this yeast in deficient media.     

Biological availability and turnover rate of acetate in marine and estuarine sediments in relation to dissimilatory sulphate reduction

Abstract Acetate turnover rates were determined using ¹⁴C acetate in sediment slurries from two Scottish sea lochs and an estuary which had different rates of oxygen uptake and sulphate reduction. Turnover rates in Loch Etive and Loch Eil were 0.504 and 0.651 μMh⁻¹ respectively, but in the River Tay Estuary there was substantially higher acetate turnover (12.22 μMh⁻¹). The addition of 20 mM sodium molybdate, a specific metabolic inhibitor of sulphate-reducing bacteria (SRB), resulted in a complete inhibition of acetate turnover. These data suggest that SRB were solely responsible for acetate oxidation in these sediments. A comparison of acetate turnover rates in the absence of molybdate and accumulation rate in the presence of the inhibitor demonstrated that at least two pools of acetate with different biological availabilities existed. In Loch Etive only 19% of chemically measured acetate was available with corresponding values of 48% and 65% for Loch Eil and the Tay Estuary respectively.     

Hypophosphite oxidase fromBacillus caldolyticus

Bacillus caldolyticus can utilize phosphorus either as phosphate, phosphite, or hypophosphite. When cultures are supplied with PO₂ as the sole source of phosphorus, the hypophosphite is oxidized to phosphate, which accumulates in the medium prior to the beginning of the log phase, and is then metabolised during growth. Resting cell suspensions also have the ability to oxidise PO₂ to PO₄. The reaction is specific for hypophosphite: PO₃ is not oxidised to PO₄, regardless of whether the cells are grown in PO₃- or PO₂-medium. The hypophosphite oxidase works optimally between pH 7.0 to 7.5, with a temperature optimum at 75°C; theK _m for NaH₂PO₂ is 320 μM. Sonication of cells, followed by high-speed centrifugation and ammonium sulfate fractionation of the cell-free extract showed that the PO₂ oxidation, which is accompanied by the formation of NADH, requires at least three components: An ammonium sulfate fraction of the cell-free extract, the residue fraction containing the respiratory chain, and NAD as cofactor. Most probably a second cofactor, so far not characterized, is required to accomplish full activity.     

Development and application of a nutrient-diffusing bioassay for large rivers

1. Laboratory and field experiments were performed to develop and then apply a nutrient-diffusing substratum (NDS) design suitable for use in large, fast-flowing rivers. 2. Initial laboratory experiments quantified diffusion of PO₄ and NO₃ from new and previously used clay pots, which were soaked in deionized distilled water. Mean release rates initially exceeded 2.4 and 725 μmol l^–1 day^–1 P and 0.22 and 18 μmol l^–1 day^–1 N from new and used pots, respectively, but declined rapidly with increasing time spent in deionized distilled water and were below detectable levels after about 18 and 29 days, respectively. 3. A phosphorus (P) dose–response experiment in a P-limited reach of the Athabasca River, Alberta, Canada showed that epilithic biomass and macroinvertebrate density on NDS increased with increasing concentrations of KH₂PO₄ up to about 0.5 m . Beyond this threshold, biomasses and densities were unaffected by initial KH₂PO₄ concentration. Coefficients of variation of epilithic biomass estimates declined with increasing KH₂PO₄ whereas invertebrate density appeared to be unaffected by KH₂PO₄ levels. 4. Release rates of both P and N from NDS filled with 0.5 m KH₂PO₄ or 0.5 m NaNO₃ declined at a log-negative rate from about 5000 μmol N-NO₃ l^–1 day^–1 and 3500 μmol P-PO₄ l^–1 day^–1 on day 2, to 200 μmol l^–1 day^–1 for both N and P on day 32. 5. After development, we used the diffusing substrata to identify spatial patterns in nutrient limitation at seven sites along a 120 km reach in the Athabasca River, that receives two known point-source nutrient inputs. NDS consisting of N, P, N + P and unenriched controls were attached to the river bottom for 22–23 days and then retrieved and sampled for epilithic chlorophyll a. Physicochemical parameters and epilithic biomasses on upper stone surfaces were also quantified when NDS were deployed and retrieved from each site. 6. Sites located immediately downstream of the two point source inputs had higher water column concentrations of PO₄ and epilithic biomasses than the site immediately upstream; epilithic biomass was positively related to PO₄ in the late autumn (r²⁼ 0.58) but not in early autumn. Sites located immediately below nutrient inputs were not nutrient-limited, whereas upstream reference sites were P-limited.     

High-performance liquid chromatographic determination of levodropropizine in human plasma with fluorometric detection

The present describes a new high-performance liquid chromatographic method with fluorescence detection for the analysis of levodropropizine [S-(−)-3-(4-phenylpiperazin-1-yl)-propane-1,2-diol] (Levotuss), an anti-tussive drug, in human serum and plasma. A reversed-phase separation of levodropropizine was coupled with detection of the native fluorescence of the molecule, using excitation and emission wavelengths of 240 nm and 350 nm respectively. The analytical column was packed with spherical 5 μm poly(styrene-divinylbenzene) particles and the mobile phase was 0.1 M NaH₂PO₄ pH 3-methanol (70:30, v/v), containing 0.5% (v/v) tetrahydrofuran. For quantitation, p-methoxylevodropropizine was used as the internal standard. Samples of 200 μl of either serum or plasma were mixed with 200 μl of 0.1 M Na₂HPO₄ pH 8.9 and extracted with 5 ml of chloroform-2-propanol (9:1, v/v). The dried residue from the organic extract was redissolved with distilled water and directly injected into the chromatograph. The limit of detection for levodropropizine, in biological matrix, was about 1–2 ng/ml, at a signal-to-noise ratio of 3. The linearity was satisfactory over a range of concentrations from 3 to 1000 ng/ml (r² = 0.99910); within-day precision tested in the range 5–100 ng/ml as well as day-to-day reproducibility proved acceptable, with relative standard deviations better than 1% in most cases. Interferences from as many as 91 therapeutic or illicit drugs were excluded.