Does weed suppression by high crop density depend on crop spatial pattern and soil water availability?

Size-asymmetric competition, in which larger plants obtain a disproportionally larger share of contested resources, can be applied in agriculture to suppress weeds by increasing crop density and spatial uniformity, as these practices enhance the initial size-asymmetric competitive advantages of crop seedlings over weed seedlings early in the growing season. We do not yet know how agronomic factors influence weed suppression at high crop density. We performed a field experiment to ask how crop density, spatial pattern and irrigation interact to influence weed suppression and grain yield in semi-arid croplands. The experimental was a factorial design with 4 factors: wheat cultivar (Ningchun4, Xihan2), irrigation level (control, irrigated), sowing density (low, 196 seeds m⁻²; moderate, 400 seeds m^-2; high, 625 seeds m⁻²), and spatial sowing pattern (rows, uniform). Weed growth was effectively suppressed by increased crop density and spatial uniformity. Effects of crop density on weed suppression and grain yield were more pronounced in the uniform pattern than in crop rows. Weed biomass was 55.7% lower and grain yield increased 29.7% higher in the high density uniform pattern compared to the low density and row pattern. Crop density interacted with cultivar in determining both weed biomass and grain yield, potentially reflecting different traits regulating crop competitive ability. Irrigation and crop density had additive effects on weed biomass but interacted to influence grain yield. Our findings support the idea that increased crop density and spatial uniformity can make a valuable and environmentally friendly contribution to weed control in wheat, reducing the need for chemical or mechanical weed control. We need a better understanding of the interactions among climate, agricultural management and crop genotype to improve our ability to effectively suppress weeds with high crop density and spatial uniformity.     

Spatial–temporal variation in soil respiration in an oak–grass savanna ecosystem in California and its partitioning into autotrophic and heterotrophic components

The spatial upscaling of soil respiration from field measurements to ecosystem levels will be biased without studying its spatial variation. We took advantage of the unique spatial gradients of an oak–grass savanna ecosystem in California, with widely spaced oak trees overlying a grass layer, to study the spatial variation in soil respiration and to use these natural gradients to partition soil respiration according to its autotrophic and heterotrophic components. We measured soil respiration along a 42.5 m transect between two oak trees in 2001 and 2002, and found that soil respiration under tree canopies decreased with distance from its base. In the open area, tree roots have no influence on soil respiration. Seasonally, soil respiration increased in spring until late April, and decreased in summer following the decrease in soil moisture content, despite the further increase in soil temperature. Soil respiration significantly increased following the rain events in autumn. During the grass growing season between November and mid-May, the average of CO₂ efflux under trees was 2.29 μmol m⁻² s⁻¹, while CO₂ efflux from the open area was 1.40 μmol m⁻² s⁻¹. We deduced that oak root respiration averaged as 0.89 μmol m⁻² s⁻¹, accounting for 39% of total soil respiration (oak root + grass root + microbes). During the dry season between mid-May and October, the average of CO₂ efflux under trees was 0.87 μmol m⁻² s⁻¹, while CO₂ efflux from the open areas was 0.51 μmol m⁻² s⁻¹. Oak root respiration was 0.36 μmol m⁻² s⁻¹, accounting for 41% of total soil respiration (oak root + microbes). The seasonal pattern of soil CO₂ efflux under trees and in open areas was simulated by a bi-variable model driven by soil temperature and moisture. The diurnal pattern was influenced by tree physiology as well. Based on the spatial gradient of soil respiration, spatial analysis of crown closure and the simulation model, we spatially and temporally upscaled chamber measurements to the ecosystem scale. We estimated that the cumulative soil respiration in 2002 was 394 gC m⁻² year⁻¹ in the open area and 616 gC m⁻² year⁻¹ under trees with a site-average of 488 gC m⁻² year⁻¹.     

Characteristics and mechanisms of the hydroponic bio-filter method for purification of eutrophic surface water

The hydroponic bio-filter method (HBFM) was adopted to purify eutrophic surface water. The average removal efficiency of total nitrogen (TN) and total phosphorus (TP) was 16.8% and 30.8%, respectively, at the hydraulic loading rate (HLR) of 3.0 m³ (m² d)⁻¹. The mass removal rate of TN and TP accordingly reached 1.0 and 0.1 g (m² d)⁻¹ separately. The sedimentation of particulate nitrogen and phosphorus played a major role in removal of nitrogen and phosphorus, which contribute 62.2% and 75.9%, respectively. The optimal HLR of HBFM ranged from 3.0 to 4.0 m³ (m² d)⁻¹. The sediment in midstream reached a maximum nitrification potential of 4.76 × 10⁻⁶ g (g h)⁻¹, while upstream it reached a maximum denitrification potential of 8.1 × 10⁻⁷ g (g h)⁻¹. The distribution of nitrification potential corresponded to the ammonium-oxidizing bacteria density. The key for improving nitrogen removal efficacy of HBFM system consisted of changing the nitrification/denitrification region distribution and accordingly enhancing the denitrification process. The sum of dissolved nitrogen removed by denitrification and plant assimilation was nearly equal to the amount released by sediment. Secateur length of Nasturtium officinale had some effect on its uptake rate. The length of cut should be less than 10 cm at a time. The harvesting frequency of once a month for N. officinale had no influence on nitrogen and phosphorus removal.     

Investigation of biomass and lipids production with Neochloris oleoabundans in photobioreactor

The fresh water microalga Neochloris oleoabundans was investigated for its ability to accumulate lipids and especially triacylglycerols (TAG). A systematic study was conducted, from the determination of the growth medium to its characterization in an airlift photobioreactor. Without nutrient limitation, a maximal biomass areal productivity of 16.5 g m⁻² day⁻¹ was found. Effects of nitrogen starvation to induce lipids accumulation was next investigated. Due to initial N. oleoabundans total lipids high content (23% of dry weight), highest productivity was obtained without mineral limitation with a maximal total lipids productivity of 3.8 g m⁻² day⁻¹. Regarding TAG, an almost similar productivity was found whatever the protocol was: continuous production without mineral limitation (0.5 g m⁻² day⁻¹) or batch production with either sudden or progressive nitrogen deprivation (0.7 g m⁻² day⁻¹). The decrease in growth rate reduces the benefit of the important lipids and TAG accumulation as obtained in nitrogen starvation (37% and 18% of dry weight, respectively).     

Effects of dwarf bamboo (Fargesia denudata) density on biomass, carbon and nutrient distribution pattern

Dense dwarf bamboo population is a structurally and functionally important component in many subalpine forest systems. To characterize the effects of stem density on biomass, carbon and majority nutrients (N, P, K, Ca and Mg) distribution pattern, three dwarf bamboo (Fargesia denudata) populations with different stem densities (Dh with 220 ± 11 stems m^?2, Dm with 140 ± 7 stems m^?2, and Dl with 80 ± 4 stems m^?2, respectively) were selected beneath a bamboo-fir (Picea purpurea) forest in Wanglang National Nature Reserve, Sichuan, China. Leaf, branch, rhizome, root and total biomass of dwarf bamboo increased with the increase of stem density, while carbon and nutrient concentrations in bamboo components decreased. Percentages of below-ground biomass and element stocks to total biomass and stocks decreased with the increase of stem density, whereas above-ground biomass and element stocks exhibited the opposite tendency. Moreover, more above-ground biomass and elements were allocated to higher part in the higher density population. In addition, percentages of culm biomass, above-ground biomass and element stocks below 100 cm culm height (H₁₀₀) increased with the increase of stem density, while percentages of branch and leaf biomass below H₁₀₀ decreased. Pearson’s correlation analyses revealed that root biomass, above-ground biomass, below-ground biomass and total biomass significantly correlated to leaf biomass in H_100?200 and total leaf biomass within high density population, while they significantly correlated to leaf biomass in H_50?150 within low density population. The results suggested that dwarf bamboo performed an efficient adaptive strategy to favor limited resources by altering biomass, carbon and nutrients distribution pattern in the dense population.     

The effect of seasonality on oxidative metabolism in Nacella (Patinigera) magellanica

We studied the seasonal variation on aerobic metabolism and the response of oxidative stress parameters in the digestive glands of the subpolar limpet Nacella (P.) magellanica. Sampling was carried out from July (winter) 2002 to July 2003 in Beagle Channel, Tierra del Fuego, Argentina. Whole animal respiration rates increased in early spring as the animals spawned and remained elevated throughout summer and fall (winter: 0.09 ± 0.02 μmol O₂ h^− 1 g^− 1; summer: 0.31 ± 0.06 μmol O₂ h^− 1 g^− 1). Oxidative stress was assessed at the hydrophilic level as the ascorbyl radical content / ascorbate content ratio (A / AH⁻). The A / AH⁻ ratio showed minimum values in winter (3.7 ± 0.2 10^− 5 AU) and increased in summer (18 ± 5 10^− 5 AU). A similar pattern was observed for lipid radical content (122 ± 29 pmol mg^− 1 fresh mass [FW] in winter and 314 ± 45 pmol mg^− 1 FW in summer), iron content (0.99 ± 0.07 and 2.7 ± 0.6 nmol mg^− 1 FW in winter and summer, respectively) and catalase activity (2.9 ± 0.2 and 7 ± 1 U mg^− 1 FW in winter and summer, respectively). Since nitrogen derived radicals are thought to be critically involved in oxidative metabolism in cells, nitric oxide content was measured and a significant difference in the content of the Fe–MGD–NO adduct in digestive glands from winter and summer animals was observed. Together, the data indicate that both oxygen and nitrogen radical generation rates in N. (P.) magellanica are strongly dependent on season.     

Assessing edge effect on the spatial distribution of selected forest biochemical properties derived using the Worldview data in Dukuduku forests,South Africa

This work explores the potential of the high‐resolution WorldView‐2 sensor in quantifying edge effects on the spatial distribution of selected forest biochemical properties in fragmented Dukuduku forest in KwaZulu‐Natal, South Africa. Specifically, we sought to map fragmented patches within forested areas in Dukuduku area, using very high spatial resolution WorldView‐2 remotely sensed data and to statistically determine the effect of these fragmented patches on the spatial distribution of selected forest biochemical properties. Edge effects on carbon, LAI and foliar nitrogen were quantified based on the models derived by Omer et al. (IEEE Journal of Selected Topics in Applied Earth Observations and Remote Sensing, 2015, 8, 4825). Edge effect statistical results on the spatial distribution of carbon, LAI and nitrogen showed significant (α = 0.05) variations with change in distance from fragmented patches (>150 m²). Forest foliar carbon concentrations significantly (p‐value = 0.016) increased from 44.8% to 45.3% with increasing distance (25–375 m) from fragmented patches. A similar trend was observed for LAI. Nevertheless, for nitrogen the results show that its concentration significantly (p = 0.016) decreased with increase in distance from the fragmented patches. Overall, the findings of this work underscore the invaluable potential and strength of WorldView‐2 data set in assessing edge effect on the spatial distribution of selected forest biochemical properties.     

Primary production and nutrient budgets of Sarcocornia perennis ssp. alpini (Lag.) Castroviejo in the salt marsh of the Palmones River estuary (Southern Spain)

Biomass, primary production and nutrient budgets associated to Sarcocornia perennis subspecies (ssp.) alpini were studied in the Palmones River estuary salt marsh (Southern Spain) to evaluate the nutrient sequestration capacity of the low marsh. Above- and belowground living and dead biomass, as well as carbon, nitrogen and phosphorus content were monitored during 1 year. Additionally, the fate of aboveground detritus was evaluated in an experiment on litter decomposition. The detritus production of S. perennis ssp. alpini was almost equivalent to its annual primary production indicating a rapid turnover of biomass. We calculated that only 12% of the aboveground detritus was exported out of the low marsh while the rest was decomposed in the sediment with a rate of 0.8 year⁻¹. Changes in concentrations of total carbon, nitrogen and phosphorus in the sediment showed patterns related to S. perennis ssp. alpini belowground biomass. Our results suggested that the sediment functions as a net sink for nutrients accumulating 550 g C m⁻² year⁻¹, 55 g N m⁻² year⁻¹, and 13 g P m⁻² year⁻¹.     

Effect of 28-homobrassinolide on antioxidant capacity and photosynthesis in Brassica juncea plants exposed to different levels of copper

A sand culture experiment was conducted to study ameliorative role of 28-homobrassinolide (HBL) in Brassica juncea seedlings raised from the seeds treated with water, or 10⁻¹⁰, 10⁻⁸ and 10⁻⁶ M 28-homobrassinolide (HBL) and grown in the presence of copper (50, 100 and 150 mg kg⁻¹ sand) and sampled at 30 days after sowing. The plants grown in the presence of copper exhibited a significant decline in growth, chlorophyll and photosynthetic parameters. However, the activity of antioxidant enzymes: catalase (E.C. 1.11.1.6), peroxidase (E.C. 1.11.1.7) and superoxide dismutase (E.C. 1.15.1.1) and the content of proline increased in the plants grown under copper stress and/or raised from treatment with HBL. However, H₂O₂ content increased significantly in copper-treated plants and decreased in plants given HBL treatment. Treatment of seeds with HBL improved the growth, photosynthetic parameters and antioxidant enzymes and also improved in the plants grown under copper stress. The elevated antioxidant enzyme and proline might be responsible to overcome the toxic effects of copper in B. juncea.     

Biomass and photosynthetic productivity of water hyacinth (Eichhornia crassipes) as affected by nutrient supply and mirid (Eccritotarus catarinensis) biocontrol

The biological control of water hyacinth is affected by water nitrogen and phosphorus content and this was investigated experimentally at five levels of nutrient supply by measuring plant photosynthetic and growth responses, and mirid reproduction and herbivory of nutrient treated plants. Low nitrogen (2–0.2 mg L⁻¹) and phosphorus (0.2–0.01 mg L⁻¹) supply decreased hyacinth photosynthesis, growth and biomass accumulation relative to plants supplied 200 mg L⁻¹ N and 20 mg L⁻¹ P. This effect depended more on nitrogen supply than phosphorus supply. Chlorophyll fluorescence showed that the photosynthetic light reactions of low nutrient plants were affected and leaves had decreased chlorophyll content, density of functional photosystems II and dissipated a greater proportion of absorbed energy as heat. Gas exchange parameters showed reduced carboxylation efficiency, rates of RuBP regeneration and light saturated photosynthetic rates, but not quantum yields. Effects on photosynthesis translated into lower plant dry biomass. Mirid herbivory exacerbated the effects of low nutrients noted for chlorophyll fluorescence, gas exchange parameters and biomass accumulation, however, these effects were not always significant and there was no obvious correlation between the level of nutrients supplied and the effect of mirid herbivory. Low nutrient supply did, however, affect mirid performance reducing the number of adult insects, nymphs and herbivory intensity suggesting that in the long-term mirid populations would be significantly affected by water nutrient status.     

Nitrogen utilization by the raphidophyte Heterosigma akashiwo: Growth and uptake kinetics in laboratory cultures

The nitrogen uptake and growth capabilities of the potentially harmful, raphidophycean flagellate Heterosigma akashiwo (Hada) Sournia were examined in unialgal batch cultures (strain CCMP 1912). Growth rates as a function of three nitrogen substrates (ammonium, nitrate and urea) were determined at saturating and sub-saturating photosynthetic photon flux densities (PPFDs). At saturating PPFD (110 μE m⁻² s⁻¹), the growth rate of H. akashiwo was slightly greater for cells grown on NH₄⁺ (0.89 d⁻¹) compared to cells grown on NO₃⁻ or urea, which had identical growth rates (0.82 d⁻¹). At sub-saturating PPFD (40 μE m⁻² s⁻¹), both urea- and NH₄⁺-grown cells grew faster than NO₃⁻-grown cells (0.61, 0.57 and 0.46 d⁻¹, respectively). The N uptake kinetic parameters were investigated using exponentially growing batch cultures of H. akashiwo and the ¹⁵N-tracer technique. Maximum specific uptake rates (V_max) for unialgal cultures grown at 15 °C and saturating PPFD (110 μE m⁻² s⁻¹) were 28.0, 18.0 and 2.89 × 10⁻³ h⁻¹ for NH₄⁺, NO₃⁻ and urea, respectively. The traditional measure of nutrient affinity—the half saturation constants (K_s) were similar for NH₄⁺ and NO₃⁻ (1.44 and 1.47 μg-at N L⁻¹), but substantially lower for urea (0.42 μg-at N L⁻¹). Whereas the α parameter (α = V_max/K_s), which is considered a more robust indicator for substrate affinity when substrate concentrations are low (<K_s), were 19.4, 12.2 and 6.88 × 10⁻³ h⁻¹/(μg-at N L⁻¹) for NH₄⁺, NO₃⁻ and urea, respectively. These laboratory results demonstrate that at both saturating and sub-saturating N concentrations, N uptake preference follows the order: NH₄⁺ > NO₃⁻ > urea, and suggests that natural blooms of H. akashiwo may be initiated or maintained by any of the three nitrogen substrates examined.     

Watershed modelling of nonpoint nitrogen losses from arable land to the Swedish coast in 1985 and 1994

Eutrophication problems in the Baltic Sea have drawn attention to the contribution of nutrients from surrounding countries. By using the HBV-N model in southern Sweden (145 000 km²) daily nitrogen leaching, reduction in rivers and lakes, net transport to the sea and source apportionment have been calculated in 3725 subbasins for the period 1985–1994, with calibration at 722 sites against measured time series. On average, 48% of the nonpoint losses from agriculture were reduced during the transport towards the sea, which left about 33 500 tonnes in annual mean net transport. This represents 45% of the total land-based load. Land cover and emissions for the years of 1985 and 1994 were used in two separate simulations of the 10-year period. The normalized gross leakage from arable land in 1985 was estimated to 29 kg N ha⁻¹ year⁻¹, which corresponds to 15 kg N ha⁻¹ year⁻¹ in net leakage to the sea. In 1994 these transports were reduced by 20 and 15%, and thereby the total load on the sea was decreased by 7%. This is still far from the Swedish goal of 50% reduction. The article presents the spatial variation of nitrogen leakage and retention within the southern half of Sweden, and emphasizes the importance of allocating measures where down-stream retention is low in order to achieve efficiency with respect to the sea. It is shown that the model approach may be used in the decision making process for best management practices in watersheds.     

Nitrogenous preference of toxigenic Pseudo-nitzschia australis (Bacillariophyceae) from field and laboratory experiments

Field and laboratory experiments were designed to determine the differential growth and toxin response to inorganic and organic nitrogen additions in Pseudo-nitzschia spp. Nitrogen enrichments of 50 μM nitrate (KNO₃), 10 μM ammonium (NH₄Cl), 20 μM urea and a control (no addition) were carried out in separate carboys with seawater collected from the mouth of the San Francisco Bay (Bolinas Bay), an area characterized by high concentrations of macronutrients and iron. All treatments showed significant increases in biomass, with chlorophyll a peaking on days 4–5 for all treatments except urea, which maintained exponential growth through the termination of the experiment. Pseudo-nitzschia australis Frenguelli abundance was 10³ cells l⁻¹ at the start of the experiment and increased by an order of magnitude by day 2. Particulate domoic acid (pDA) was initially low but detectable (0.15 μg l⁻¹), and increased throughout exponential and stationary phases across all treatments. At the termination of the experiment, the urea treatment produced more than double the amount of pDA (9.39 μg l⁻¹) than that produced by the nitrate treatment (4.26 μg l⁻¹) and triple that of the control and ammonium treatments (1.36 μg l⁻¹ and 2.64 μg l⁻¹, respectively). The mean specific growth rates, calculated from increases in chlorophyll a and from cellular abundance of P. australis, were statistically similar across all treatments.These field results confirmed laboratory experiments conducted with a P. australis strain isolated from Monterey Bay, CA (isolate AU221-a) grown in artificial seawater enriched with 50 μM nitrate, 50 μM ammonium or 25 μM of urea as the sole nitrogen source. The exponential growth rate of P. australis was significantly slower for cells grown on urea (ca. 0.5 day⁻¹) compared to the cells grown on either nitrate or ammonium (ca. 0.9 day⁻¹). However the urea-grown cells produced more particulate and dissolved domoic acid (DA) than the ammonium- or nitrate-grown cells. The field and laboratory experiments demonstrate that P. australis is able to grow effectively on urea as the primary source of nitrogen and produced more pDA when grown on urea in both natural assemblages and unialgal cultures. These results suggest that the influence of urea from coastal runoff may prove to be more important in the development or maintenance of toxic blooms than previously thought, and that the source of nitrogen may be a determining factor in the relative toxicity of west coast blooms of P. australis.     

Influence of bed media characteristics on ammonia and nitrate removal in shallow horizontal subsurface flow constructed wetlands

Two bed media were tested (gravel and Filtralite) in shallow horizontal subsurface flow (HSSF) constructed wetlands in order to evaluate the removal of ammonia and nitrate for different types of wastewater (acetate-based and domestic wastewater) and different COD/N ratios. The use of Filtralite allowed both higher mass removal rates (1.1 g NH₄–N m⁻² d⁻¹ and 3 g NO₃–N m⁻² d⁻¹) and removal efficiencies (>62% for ammonia, 90–100% for nitrate), in less than 2 weeks, when compared to the ones observed with gravel. The COD/N ratio seems to have no significant influence on nitrate removal and the removal of both ammonia and nitrate seems to have involved not only the conventional pathways of nitrification–denitrification. The nitrogen loading rate of both ammonia (0.8–2.4 g NH₄–N m⁻² d⁻¹) and nitrate (0.6–3.2 g NO₃–N m⁻² d⁻¹) seem to have influenced the respective removal rates.     

Nutrient runoff dynamics in a rural catchment: Influence of land-use changes, climatic fluctuations and ecotechnological measures

The main trend in land-use changes in the Porijõgi River catchment, south Estonia, is a significant increase in abandoned lands (from 1.7% in 1987 to 10.5% in 1997), and a decrease in arable lands (from 41.8 to 23.9%). Significant climatic fluctuations occurred during the last decades. Milder winters (increase of air temperature in February from −7.9 to −5.5°C during the period 1950–1997) and a change in the precipitation pattern have influenced the mean annual water discharge. This results in more intensive material flow during colder seasons and decreased water runoff in summer. During the period 1987–1997 the runoff of total-N, total-P, SO₄, and organic material (after BOD₅) decreased from 25.9 to 5.1, from 0.32 to 0.13, from 78 to 48, and from 7.4 to 3.5 kg ha⁻¹ year⁻¹, respectively. Most significant was a 4–20-fold decrease in agricultural subcatchments while in the forested upper-course catchment the changes were insignificant. Variations of total-N, and total-P runoff in both the entire catchment and its agricultural subcatchments are well described by the change of land use (including fertilization intensity), soil parameters and water discharge. In small agricultural subcatchments the rate of fertilization was found to be the most important factor affecting nitrogen runoff, while land-use pattern plays the main role in larger mosaic catchments. Ecotechnological measures (e.g. riparian buffer zones and buffer strips, constructed wetlands) to control nutrient flows from agricultural catchments are very important.     

Variation in domoic acid concentration in king scallop (Pecten maximus) from fishing grounds around the Isle of Man

Domoic acid (DA), the toxin responsible for amnesic shellfish poisoning (ASP) can accumulate in king scallop Pecten maximus leading to extensive fishery closures. Approximately 59% of the total value of all fish and shellfish landed in the Isle of Man in 2004 comprised king scallop, hence the economy of the Manx marine sector is particularly susceptible to impacts from this biotoxin. Scallop from fishing grounds around the Isle of Man were sampled in October 2003, June 2004 and October 2004 to determine levels of inter-animal and spatial variability in DA concentration and factors that might influence toxin concentration such as scallop size and water depth. Mean DA concentrations in hepatopancreas ranged from 296.3 μg g⁻¹ to below the detection limit, in gonad from 27.8 μg g⁻¹ to below the limit of detection and in adductor muscle from 7.3 μg g⁻¹ to below the limit of detection. High levels of inter-animal variability of DA concentration in hepatopancreas were recorded; CVs ranging from 16.1% to 70.0%. DA concentrations above 20 μg g⁻¹ were recorded in gonads on all three sampling dates. Scallops from fishing grounds on the east of the Isle of Man were significantly less contaminated than those from the west and southwest. A significant positive correlation between DA concentration and shell length was recorded in some sites, but there was no relationship with water depth. The high inter-animal, spatial and seasonal variability in toxin concentration highlighted the importance of understanding field variability for the development of reliable sampling and management protocols.     

Interactions between Prorocentrum donghaiense Lu and Scrippsiella trochoidea (Stein) Loeblich III under laboratory culture

Interactions between Prorocentrum donghaiense Lu and Scrippsiella trochoidea (Stein) Loeblich III, two species of causative bloom dinoflagellates in China, were investigated using bi-algal cultures under controlled laboratory conditions. The growth of P. donghaiense and S. trochoidea were significantly suppressed when the initial cell densities were set at 1.9 × 10⁴ cells mL⁻¹ or 1.9 × 10⁵ cells mL⁻¹ for P. donghaiense and 1.0 × 10⁴ cells mL⁻¹ for S. trochoidea when the initial size/density ratio was 1:1 or 10:1, respectively, but no out-competement was observed in either bi-algal culture by the end. The simultaneous assay on the culture filtrate showed that P. donghaiense filtrate prepared at a lower initial density (1.9 × 10⁴ cells mL⁻¹) stimulated the co-cultured S. trochoidea at a density of 1.0 × 10⁴ cells mL⁻¹, but filtrate at a higher density (1.9 × 10⁵ cells mL⁻¹) depressed its growth. Differently, the filtrate of S. trochoidea at a density of 1.0 × 10⁴ cells mL⁻¹ significantly suppressed the growth of P. donghaiense at a density of 1.9 × 10⁴ cells mL⁻¹, but had little stimulatory effect on P. donghaiense at a density of 1.9 × 10⁵ cells mL⁻¹compared to the control (P > 0.05). It is likely that these two species of microalgae interact with each other mainly by releasing allelochemical substance(s) into the culture medium, and a direct cell-to-cell contact was not necessary for their mutual interaction. We then quantify their interactions in the bi-algal culture by using a mathematical model. The estimated parameters from the model showed that the inhibition exerted by S. trochoidea on P. donghaiense was about 43 and 24 times stronger than the inhibitory effect that P. donghaiense exerted on S. trochoidea when the initial size/density were 1:1 and 10:1, respectively. S. trochoidea seemed to have a survival strategy that was superior to P. donghaiense in the bi-algal culture under controlled laboratory conditions. We also observed a closely positive relationship between the initial cell density and its effect on the co-cultured microalga by measuring the fluorenscence: filtrate prepared from higher initial cell density had stronger interference on the co-cultured microalga. Moreover, pre-treated under different temperature conditions (30 °C, 60 °C and 100 °C) would significantly changed the effect of culture filtrate on the co-cultured microalga. Result inferred that P. donghaiense or S. trochoidea would release allelochemicals into the bi-algal culture medium and the allelochemicals might be a mixture with temperature-sensitive components in it.     

Influence of humic, fulvic and hydrophilic acids on the growth, photosynthesis and respiration of the dinoflagellate Prorocentrum minimum (Pavillard) Schiller

Blooms of the dinoflagellate Prorocentrum minimum often occur in coastal regions characterized by variable salinity and elevated concentrations of terrestrially derived dissolved organic carbon (DOC). Humic, fulvic and hydrophilic acid fractions of DOC were isolated from runoff entering lower Narragansett Bay immediately after a rainfall event and the influence of these fractions upon P. minimum growth, cell yield, photosynthesis and respiration was examined. All organic fractions stimulated growth rates and cell yields compared with controls (no organic additions), but the extent of stimulation varied with the fraction and its molecular weight. Greatest stimulations were observed with humic and fulvic acids additions; cell yields were more than 2.5 and 3.5 times higher than with hydrophilic acid additions while growth rates were 21 and 44% higher, respectively. Responses to additions of different molecular weight fractions of each DOC fraction suggest that growth rate effects were attributable to specific molecular weight fractions: the >10,000 fraction of humic acids, both the >10,000 and <500 fractions of fulvic acids and the <10,000 fraction of hydrophilic acids. The form and concentration of nitrogen (as NO₃⁻ or NH₄⁺) present also influenced P. minimum response to DOC; 10–20 μg ml⁻¹ additions of fulvic acid had no effect upon growth rates in the presence of NH₄⁺ but significantly increased growth rates in the presence of NO₃⁻, a relationship probably related to fulvic acid effects upon trace metal bioavailability and subsequent regulation of the biosynthesis of enzymes required for NO₃⁻ assimilation. The influence of DOC additions on P. minimum respiration and production rates also varied with the organic fraction and its concentration. Production rates ranged from 1.1 to 3.4 pg O₂ cell⁻¹ h⁻¹, with highest rates observed upon exposure to fulvic and hydrophilic acid concentrations of >10 μm ml⁻¹. Low concentrations (5–10 μg ml⁻¹) of humic acid had no statistically significant effect upon production, but exposure to concentrations >25 μg ml⁻¹ resulted in a 30% decrease in O₂ evolution, probably due to light attenuation by the highly colored humic acid fraction. Respiration rates ranged from 1.2 to 2.7 pg O₂ cell⁻¹ h⁻¹ and were elevated upon exposure to both fulvic and hydrophilic acids, but not to humic acid. These results demonstrate that terrestrially derived DOC fractions play an active role in stimulation of P. minimum growth via direct effects upon growth, yield and photosynthesis as well as via indirect influences such as interactions with nitrogen and effects upon light attenuation.     

Nitrogen source effects on the growth and toxicity of two strains of the ciguatera-causing dinoflagellate Gambierdiscus toxicus

Two Caribbean strains (1651 and 1655) of the ciguatera-causing dinoflagellate Gambierdiscus toxicus were grown in xenic, batch culture under defined, measured nutrient conditions with nitrate, ammonium, urea, a mix of free amino acids (FAA), or putrescine as the nitrogen source. Cultures were maintained at 27 °C, salinity 35, 110 μmol m⁻² s⁻¹ (12 h:12 h light:dark cycle) on L2 medium at an initial nitrogen concentration of 50 μM N. Toxicity was determined using a ouabain/veratridine-dependent cytotoxicity assay (N2A assay) standardized to a ciguatoxin standard. Nitrate, ammonium, FAA, and putrescine supported growth, but urea did not. The appearance of ammonium in the organic nitrogen cultures indicated that G. toxicus and/or associated bacteria remineralized the available organic nitrogen. Both strains were exposed to nitrogen-limiting conditions as evidenced by chlorophyll a content per cell, nitrogen content, and nitrogen (N) to phosphorus (P) (N:P) ratio significantly declining once nitrogen was no longer available in the medium and cells entered stationary phase. Strain 1651 grew significantly faster than strain 1655 when nitrate, FAA, and putrescine was the nitrogen source, but not ammonium. Nitrogen source had no effect on growth rate (0.14 d⁻¹) in strain 1651. The growth rate of strain 1655 (0.10–0.13 d⁻¹) was significantly faster on ammonium than the other nitrogen sources. Strain 1655 was significantly more toxic (10-fold) than strain 1651 except when growing on ammonium at exponential phase. Toxicity ranged from 1.3 to 8.7 fg C-CTX1-Eq cell⁻¹ in strain 1651 and from 30.7 to 54.3 fg C-CTX1-Eq cell⁻¹ in strain 1655. Nitrogen source had no significant affect on toxicity. Toxicity was greater in stationary versus exponential phase cells for strain 1651 when grown on nitrate and strain 1655 regardless of nitrogen source. The difference in toxicity between growth phases may result from an increase in ciguatoxin and/or maitotoxin. Our results suggest that some strains of G. toxicus when associated with bacteria are able to take advantage of organic as well as inorganic nitrogen sources on short time scales to support future growth. The uncoupling of total nitrogen and phosphorus pools from conditions in the water column suggest that instantaneous growth rates can be supported by nutrients acquired hours to days earlier.     

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.