Reactivation of aerobic and anaerobic ammonium oxidizers in OLAND biomass after long-term storage

The biomass of an oxygen-limited autotrophic nitrification/denitrification (OLAND) biofilm reactor was preserved in various ways to find a storage method for both aerobic and anaerobic ammonium-oxidizing bacteria (AerAOB and AnAOB). Storage occurred at −20°C with and without glycerol as cryoprotectant and at 4 and 20°C with and without nitrate as redox buffer. After 2 and 5 months, reactivation of AerAOB and AnAOB was achieved with the biomass stored at 4°C with and without nitrate and at 20°C with nitrate. Moreover, the presence of the AerAOB and AnAOB was confirmed with fluorescent in situ hybridization (FISH). Preservation in a nitrate environment resulted in a lag phase for the AnAOB reactivation. The supplied nitrate was denitrified during storage, and a real-time polymerase chain reaction with nitrifying and denitrifying genes allowed to estimate that at least 1.0 to 6.0% of the OLAND biofilm consisted of denitrifiers. It was concluded that reactivation after long-term storage is possible and that preservation at 4°C without nitrate addition is the recommended storage technique. The possibility to store OLAND biomass will facilitate research on AnAOB and can overcome larger-scale start-up and inhibition problems of novel nitrogen processes involving AnAOB.     

Long-chain acylhomoserine lactones increase the anoxic ammonium oxidation rate in an OLAND biofilm

The oxygen-limited autotrophic nitrification/denitrification (OLAND) process comprises one-stage partial nitritation and anammox, catalyzed by aerobic and anoxic ammonium-oxidizing bacteria (AerAOB and AnAOB), respectively. The goal of this study was to investigate whether quorum sensing influences anoxic ammonium oxidation in an OLAND biofilm, with AnAOB colonizing 13% of the biofilm, as determined with fluorescent in situ hybridization (FISH). At high biomass concentrations, the specific anoxic ammonium oxidation rate of the OLAND biofilm significantly increased with a factor of 1.5 ± 0.2 compared to low biomass concentrations. Supernatant obtained from the biofilm showed no ammonium-oxidizing activity on itself, but its addition to low OLAND biomass concentrations resulted in a significant activity increase of the biomass. In the biofilm supernatant, the presence of long-chain acylhomoserine lactones (AHLs) was shown using the reporter strain Chromobacterium violaceum CV026, and one specific AHL, N-dodecanoyl homoserine lactone (C₁₂-HSL), was identified via LC-MS/MS. Furthermore, C₁₂-HSL was detected in an AnAOB-enriched community, but not in an AerAOB-enriched community. Addition of C₁₂-HSL to low OLAND biomass concentrations resulted in a significantly higher ammonium oxidation rate (p < 0.05). To our knowledge, this is the first report demonstrating that AHLs enhance the anoxic ammonium oxidation process. Future work should confirm which species are responsible for the in situ production of C₁₂-HSL in AnAOB-based applications.     

Seasonal accumulation and partitioning of nitrogen by lentil

Although wheat (Triticum aestivum L.) is the dominant crop of the semi-arid plains of Canada and the western United States, lentil (Lens culinaris Medik.) has become an important alternative crop. Sources and seasonal accumulation of N must be understood in order to identify parameters that can lead to increased N₂-fixing activity and yield. Inoculated lentil was grown in a sandy-loam soil at an irrigated site in Saskatchewan, Canada. Wheat was used as the reference crop to estimate N₂ fixation by the A-value approach. Lentil and wheat received 10 and 100 kg N ha⁻¹ of ammonium nitrate, respectively. Crops were harvested six times during the growing season and plant components analyzed. During the first 71 days after planting the wheat had a higher daily dry matter and N accumulation compared to lentil. However, during the latter part of the growing season, daily dry matter and N accumulation were greater for lentil. The maximum total N accumulation for lentil at maturity was 149 kg ha⁻¹. In contrast, wheat had a maximum N accumulation of 98 kg ha⁻¹ in the Feekes 11.1 stage, or 86 days after planting. The maximum daily rates of N accumulation were 3.82 kg N ha⁻¹ day⁻¹ for lentil and 2.21 kg N ha⁻¹ day⁻¹ for wheat. The percentage of N derived from N₂ fixation (% Ndfa) ranged from 0 at the first harvest to 92 % at final harvest. Generative plant components had higher values for % Ndfa than the vegetative components which indicates that N in the reproductive plant parts was derived largely from current N₂ fixation and lentil continued to fix N until the end of the pod fill stage. At final harvest, lentil had derived 129 kg N ha⁻¹ from N₂ fixation with maximum N₂-fixing activity (4.4 kg N ha⁻¹ day⁻¹) occurring during the early stages of pod fill. Higher maximum rates of N₂-fixing activity than net N accumulation (3.82 kg N ha⁻¹ day⁻¹) may have been caused by N losses like volatilization. In addition, lentil provided a net N contribution to the soil of 59 kg ha⁻¹ following the removal of the grain.     

Kinetics of CO conversion into H<Subscript>2</Subscript> by <Emphasis Type="Italic">Carboxydothermus hydrogenoformans</Emphasis>

The objective of this study was to improve the biological water–gas shift reaction for producing hydrogen (H₂) by conversion of carbon monoxide (CO) using an anaerobic thermophilic pure strain, Carboxydothermus hydrogenoformans. Specific hydrogen production rates and yields were investigated at initial biomass densities varying from 5 to 20 mg volatile suspended solid (VSS) L⁻¹. Results showed that the gas–liquid mass transfer limits the CO conversion rate at high biomass concentrations. At 100-rpm agitation and at CO partial pressure of 1 atm, the optimal substrate/biomass ratio must exceed 5 mol CO g⁻¹ biomass VSS in order to avoid gas–liquid substrate transfer limitation. An average H₂ yield of 94 ± 3% and a specific hydrogen production rate of ca. 3 mol g⁻¹ VSS day⁻¹ were obtained at initial biomass densities between 5 and 8 mg VSS⁻¹. In addition, CO bioconversion kinetics was assessed at CO partial pressure from 0.16 to 2 atm, corresponding to a dissolved CO concentration at 70°C from 0.09 to 1.1 mM. Specific bioactivity was maximal at 3.5 mol CO g⁻¹ VSS day⁻¹ for a dissolved CO concentration of 0.55 mM in the culture. This optimal concentration is higher than with most other hydrogenogenic carboxydotrophic species.     

Biofiltering efficiency, uptake and assimilation rates of Ulva clathrata (Roth) J. Agardh (Clorophyceae) cultivated in shrimp aquaculture waste water

The growth, biofiltering efficiency and uptake rates of Ulva clathrata were studied in a series of outdoor tanks, receiving waste water directly from a shrimp (Litopenaeus vannamei) aquaculture pond, under constant aeration and two different water regimes: (1) continuous flow, with 1 volume exchange a day (VE day^-1) and (2) static regime, with 1 VE after 4 days. Water temperature, salinity, pH, dissolved inorganic nitrogen (DIN), phosphate (PO₄), chlorophyll-a (chl-a), total suspended solids (TSS), macroalgal biomass (fresh weight) and tissue nutrient assimilation were monitored over 12 days. Ulva clathrata was highly efficient in removing the main inorganic nutrients from effluent water, stripping 70–82% of the total ammonium nitrogen (TAN) and 50% PO₄ within 15 h. Reductions in control tanks were much lower (Tukey HSD, P < 0.05). After 3 days, the mean uptake rates by the seaweed biomass under continuous flow were 3.09 mg DIN g DW day⁻¹ (383 mg DIN m⁻² day⁻¹) and 0.13 mg PO₄ g DW day⁻¹ (99 mg PO₄ m⁻² day⁻¹), being significantly higher than in the static regime (Tukey HSD, P < 0.05). The chl-a decreased in seaweed tanks, suggesting that U. clathrata inhibited phytoplankton growth. Correlations between the cumulative values of DIN removed from the water and total nitrogen assimilated into the seaweed biomass (r = 0.7 and 0.8, P < 0.05), suggest that nutrient removal by U. clathrata dominated over other processes such as phytoplankton and bacterial assimilation, ammonia volatilization and nutrient precipitation.     

Increasing fish production from wetlands at Lake Victoria, Uganda using organically manured seasonal wetland fish ponds

The processes driving primary productivity and its impacts on fish production were investigated in field trials in eight seasonal earthen wetland ponds ‘Fingerponds’ (192 m²) in Uganda between 2003 and 2005. The ponds were stocked by the seasonal flood with predominantly Oreochromis spp. at densities ranging from 0.1 to 0.5 fish m⁻². Chicken manure (521, 833 or 1,563 kg ha⁻¹) was applied fortnightly. Results showed that primary productivity was enhanced with maximum average net primary productivity (±Standard Error) of 11.7 (±2.5) g O₂ m⁻² day⁻¹ at the Gaba site and 8.3 (±1.5) g O₂ m⁻² day⁻¹ at the Walukuba site. Net fish yields were higher in manured ponds with up to 2,670 kg ha⁻¹ yield for a 310 day growth period compared to less than 700 kg ha⁻¹ in unmanured ponds. Fish production was limited mainly by high recruitment, falling water levels, light limitation from high suspended solids and turbidity, and low zooplankton biomass. It was concluded that Fingerponds have a high potential for sustainable fish production and can contribute to the alleviation of protein shortages amongst the riparian communities around Lake Victoria. Production can be enhanced further with improved stock management.     

Nitrous oxide and methane fluxes of a pristine slope mire in the German National Park Harz Mountains

Pristine peatlands covered by Histosols (bogs and fens) with high water table and a restricted oxygen (O₂) availability are known to have low emissions of nitrous oxide (N₂O) but may be a significant source for atmospheric methane (CH₄) which are both important greenhouse gases. For the first time N₂O and CH₄ fluxes of a pristine slope mire in the German Harz Mountains have been monitored. Previously reported peatlands are characterised by anaerobic conditions due to high water table levels. Slope mires monitored here receive O₂ through slope water inflow. Gas fluxes have been monitored deploying closed chamber method on a central non-forested area and a forested area at the periphery of the slope mire. By means of groundwater piezometers water table levels, ammonium and nitrate contents as well as hydro-chemical variables like oxygen content and redox potential of the mire pore water have been concurrently measured with trace gas fluxes at both monitoring sites of the slope mire. The slope mire took up small amounts of atmospheric methane at a rate of −0.02 ± 0.01 kg C ha⁻¹ year⁻¹ revealing no significant difference between the forested and non-forested site. Higher uptake rates were observed during low water table level. In contrast to pristine peatlands influx of oxygen containing pore water into slope mire does limit reduction processes and resultant CH₄ emission. N₂O fluxes of the forested and non-forested sites of the slope mire did not differ and amounted to 0.25 ± 0.44 kg N ha⁻¹ year⁻¹. Higher emissions were observed at low water table levels and during thawing periods. In spite of favourable conditions N₂O fluxes of the slope mire have been comparable to those of pristine peatlands.     

Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method

Dinitrogen fixation in white clover (Trifolium repens L.) grown in pure stand and mixture with perennial ryegrass (Lolium perenne L.) was determined in the field using ¹⁵N isotope dilution and harvest of the shoots. The apparent transfer of clover N to perennial ryegrass was simultaneously assessed. The soil was labelled either by immobilizing ¹⁵N in organic matter prior to establishment of the sward or by using the conventional labelling procedure in which ¹⁵N fertilizer is added after sward establishment. Immobilization of ¹⁵N in the soil organic matter has not previously been used in studies of N₂ fixation in grass/clover pastures. However, this approach was a successful means of labelling, since the ¹⁵N enrichment only declined at a very slow rate during the experiment. After the second production year only 10–16% of the applied ¹⁵N was recovered in the harvested herbage. The two labelling methods gave, nonetheless, a similar estimate of the percentage of clover N derived from N₂ fixation. In pure stand clover, 75–94% of the N was derived from N₂ fixation and in the mixture 85–97%. The dry matter yield of the clover in mixture as percentage of total dry matter yield was relatively high and increased from 59% in the first to 65% in the second production year. The average daily N₂ fixation rate in the mixture-grown clover varied from less than 0.5 kg N ha⁻¹ day⁻¹ in autumn to more than 2.6 kg N ha⁻¹ day⁻¹ in June. For clover in pure stand the average N₂ fixation rate was greater and varied between 0.5 and 3.3 kg N ha⁻¹ day⁻¹, but with the same seasonal pattern as for clover in mixture. The amount of N fixed in the mixture was 23, 187 and 177 kg N ha⁻¹ in the seeding, first and second production year, respectively, whereas pure stand clover fixed 28, 262 and 211 kg N ha⁻¹ in the three years. The apparent transfer of clover N to grass was negligible in the seeding year, but clover N deposited in the rhizosphere or released by turnover of stolons, roots and nodules, contributed 19 and 28 kg N ha⁻¹ to the grass in the first and second production year, respectively. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nitrogen uptake in a Norway spruce stand following ammonium sulphate application,fertigation, irrigation,drought and nitrogen-free-fertilisation

The above-ground accumulation of N,N uptake and litter quality resulting from improved or deteriorated availability of water and nutrients in a 25 year old Norway spruce stand in SW Sweden (as part of the Skogaby project) is presented. Treatment include irrigation; artificial drought; ammonium sulphate addition; N-free-fertilisation and irrigation with liquid fertilisers including a complete set of nutrients according to the Ingested principle (fertigation). At start of the experiment the stand contained 86.5 t dry mass and 352 kg N ha⁻¹. The following three years the annual N uptake in untreated trees was 32 kg N ha⁻¹ to be compared with the annual N throughfall of 17 kg ha⁻¹. Simultaneously, the treatment with ammonium sulphate and liquid fertilisation resulted in 48 and 56 kg ha⁻¹ y⁻¹, respectively, in treatment specific N-uptake following an application of 100 kg N ha⁻¹ y⁻¹. Addition of a N-free fertiliser resulted in improved N-uptake by 19 kg N ha⁻¹ y⁻¹ and irrigation by 10 kg N ha⁻¹ y⁻¹, compared to control. A linear relation between total above-ground dry mass production and N-uptake was found for trees growing with similar water availability. Dry mass production increased with increased water availability given the same N-uptake. It is concluded that the studied stand this far is not N saturated', as N fertilisation resulted in both increased N uptake and increased growth. Addition of a N-free-fertiliser resulted in increased uptake of N compared to the control, indicating an increased mineralisation rate or uptake capacity of the root system. The linear relation between N uptake and biomass production shows that at this study site N is a highly limiting factor for growth.     

Seasonal variation in the nitrogenase activity of aPanicum maximum var.trichoglume pasture and identification of associated bacteria

Summary The influence of seasonal variation on nitrogenase (N₂-ase) activity of undisturbed soil-plant cores ofPanicum maximum var.trichoglume was measured using the C₂H₂ reduction assay. The largest N₂-ase activity in the field, 14.7 g N ha⁻¹ day⁻¹, occurred in spring when soil moisture was high, soil temperature was low and nitrogenous fertiliser influence was at a minimum. The potential N₂-ase activity of the cores, measured under controlled conditions, reached a maximum of 27.2 g N ha⁻¹ day⁻¹ and averaged 26.3 g N ha⁻¹ day⁻¹ over the 14 month sampling period. N₂-ase activity was positively correlated (P=0.05) with field soil moisture and negatively correlated with field soil temperature (r=0.59 and −0.78 respectively). Multiple regression showed that 69% of the variation of N₂-ase activity in the field was associated with the combined effects of soil moisture and soil temperature. Nitrogen fixing bacteria were isolated from the roots ofP. maximum and based upon morphology, biochemical tests and fluorescent antibody reaction, were found to be closely related toAzospirillum lipoferum.     

Biomass and aboveground net primary production in a subtropical mangrove stand of Kandelia obovata (S., L.) Yong at Manko Wetland, Okinawa, Japan

Biomass and aboveground net primary production (ANPP) in a monospecific pioneer stand of a mangrove Kandelia obovata (S., L.) Yong were quantified. The estimated biomasses in leaves, branches, stems, roots, aboveground and total were 5.61 (3.68%), 28.8 (18.9%), 46.1 (30.2%), 71.8 (47.2%), 80.5 (52.8%) and 152 Mg ha⁻¹ (100%), respectively. Stem phytomass increment per tree was estimated using allometric relationships and stem analysis. Stem volume without bark of harvested trees showed a strong allometric relationship with D _0.1² H (D _0.1, diameter at a height of one-tenth of tree height H) (R ² = 0.924). Annual stem volume increment per tree showed a strong allometric relationship with D _0.1² H (R ² = 0.860). Litterfall rate ranges from 3.87 to 56.1 kg ha⁻¹ day⁻¹ for leaves and 0.177 to 46.2 kg ha⁻¹ day⁻¹ for branches. Seasonal changes of litterfall rate were observed, which showed a peak during wet season (August–September). Total annual litterfall was estimated as 10.6 Mg ha⁻¹ year⁻¹, in which 68.2% was contributed by the leaves. The ANPP in the K. obovata stand was 29.9–32.1 Mg ha⁻¹ year⁻¹, which is ca. 2.8–3.0 times of annual litterfall. The growth efficiency (aboveground biomass increment/LAI) was 5.35–5.98 Mg ha⁻¹ year⁻¹. The low leaf longevity (9.3 months) and high growth efficiency of K. obovata makes it a highly productive mangrove species.     

Nitrogen retention in the hyporheic zone of a glacial river in interior Alaska

We examined the hydrologic controls on nitrogen biogeochemistry in the hyporheic zone of the Tanana River, a glacially-fed river, in interior Alaska. We measured hyporheic solute concentrations, gas partial pressures, water table height, and flow rates along subsurface flowpaths on two islands for three summers. Denitrification was quantified using an in situ ¹⁵NO₃⁻ push–pull technique. Hyporheic water level responded rapidly to change in river stage, with the sites flooding periodically in mid−July to early−August. Nitrate concentration was nearly 3-fold greater in river (ca. 100 μg NO₃⁻–N l⁻¹) than hyporheic water (ca. 38 μg NO₃⁻–N l⁻¹), but approximately 60–80% of river nitrate was removed during the first 50 m of hyporheic flowpath. Denitrification during high river stage ranged from 1.9 to 29.4 mg N kg sediment⁻¹ day⁻¹. Hotspots of methane partial pressure, averaging 50,000 ppmv, occurred in densely vegetated sites in conjunction with mean oxygen concentration below 0.5 mgO₂ l⁻¹. Hyporheic flow was an important mechanism of nitrogen supply to microbes and plant roots, transporting on average 0.41 gNO₃⁻–N m⁻² day⁻¹, 0.22 g NH₄⁺–N m⁻² day⁻¹, and 3.6 g DON m⁻² day⁻¹ through surface sediment (top 2 m). Our results suggest that denitrification can be a major sink for river nitrate in boreal forest floodplain soils, particularly at the river-sediment interface. The stability of the river hydrograph and the resulting duration of soil saturation are key factors regulating the redox environment and anaerobic metabolism in the hyporheic zone.     

Nitrogen dynamics and soil nitrate retention in a <Emphasis Type="Italic">Coffea arabica</Emphasis>—<Emphasis Type="Italic">Eucalyptus deglupta</Emphasis> agroforestry system in Southern Costa Rica

Nitrogen fertilization is a key factor for coffee production but creates a risk of water contamination through nitrate (NO₃⁻) leaching in heavily fertilized plantations under high rainfall. The inclusion of fast growing timber trees in these coffee plantations may increase total biomass and reduce nutrient leaching. Potential controls of N loss were measured in an unshaded coffee (Coffea arabica L.) plot and in an adjacent coffee plot shaded with the timber species Eucalyptus deglupta Blume (110 trees ha⁻¹), established on an Acrisol that received 180 kg N ha⁻¹ as ammonium-nitrate and 2,700 mm yr⁻¹ rainfall. Results of the one year study showed that these trees had little effect on the N budget although some N fluxes were modified. Soil N mineralization and nitrification rates in the 0–20 cm soil layer were similar in both systems (≈280 kg N ha⁻¹ yr⁻¹). N export in coffee harvest (2002) was 34 and 25 kg N ha⁻¹ yr⁻¹ in unshaded and shaded coffee, and N accumulation in permanent biomass and litter was 25 and 45 kg N ha⁻¹ yr⁻¹, respectively. The losses in surface runoff (≈0.8 kg mineral N ha⁻¹ yr⁻¹) and N₂O emissions (1.9 kg N ha⁻¹ yr⁻¹) were low in both cases. Lysimeters located at 60, 120, and 200 cm depths in shaded coffee, detected average concentrations of 12.9, 6.1 and 1.2 mg NO₃⁻-N l⁻¹, respectively. Drainage was slightly reduced in the coffee-timber plantation. NO₃⁻ leaching at 200 cm depth was about 27 ± 10 and 16 ± 7 kg N ha⁻¹ yr⁻¹ in unshaded and shaded coffee, respectively. In both plots, very low NO₃⁻ concentrations in soil solution at 200 cm depth (and in groundwater) were apparently due to NO₃⁻ adsorption in the subsoil but the duration of this process is not presently known. In these conventional coffee plantations, fertilization and agroforestry practices must be refined to match plant needs and limit potential NO₃⁻ contamination of subsoil and shallow soil water.     

Biological nitrification–denitrification with alternating oxic and anoxic operations using aerobic granules

Efficient nitrification and denitrification of wastewater containing 1,700 mgl⁻¹ of ammonium-nitrogen was achieved using aerobic granular sludge cultivated at medium-to-high organic loading rates. The cultivated granules were tested in a sequencing batch reactor (SBR) fed with 6.4 or 10.2 kg NH₄⁺-N m⁻³ day⁻¹, a loading significantly higher than that reported in literature. With alternating 2 h oxic and 2 h anoxic operation (OA) modes, removal rate was 45.5 mg NH₄⁺-N g⁻¹ volatile suspended solids⁻¹ h⁻¹ at 6.4 kg NH₄⁺-N m⁻³ day⁻¹ loading and 41.3 ± 2.0 at 10.2 kg NH₄⁺-N m⁻³ day⁻¹ loading. Following the 60 days SBR test, granules were intact. The fluorescence in situ hybridization and confocal laser scanning microscopy results indicate that the SBR-OA granules have a distribution with nitrifers outside and heterotrophs outside that can effectively expose functional strains to surrounding substrates at high concentrations with minimal mass transfer limit. This microbial alignment combined with the smooth granule surface achieved nitrification–denitrification of wastewaters containing high-strength ammonium using aerobic granules. Conversely, the SBR continuous aeration mode yielded a distribution with nitrifers outside and heterotrophs inside with an unsatisfactory denitrification rate and floating granules as gas likely accumulated deep in the granules.     

Microphytobenthos of Arctic Kongsfjorden (Svalbard, Norway): biomass and potential primary production along the shore line

During summer 2007, Arctic microphytobenthic potential primary production was measured at several stations around the coastline of Kongsfjorden (Svalbard, Norway) at ≤5 m water depth and at two stations at five different water depths (5, 10, 15, 20, 30 m). Oxygen planar optode sensor spots were used ex situ to determine oxygen exchange in the overlying water of intact sediment cores under controlled light (ca. 100 μmol photons m⁻² s⁻¹) and temperature (2–4°C) conditions. Patches of microalgae (mainly diatoms) covering sandy sediments at water depths down to 30 m showed high biomass of up to 317 mg chl a m⁻². In spite of increasing water depth, no significant trend in “photoautotrophic active biomass” (chl a, ratio living/dead cells, cell sizes) and, thus, in primary production was measured at both stations. All sites from ≤5 to 30 m water depth exhibited variable rates of net production from −19 to +40 mg O₂ m⁻² h⁻¹ (−168 to +360 mg C m⁻² day⁻¹) and gross production of about 2–62 mg O₂ m⁻² h⁻¹ (17–554 mg C m⁻² day⁻¹), which is comparable to other polar as well as temperate regions. No relation between photoautotrophic biomass and gross/net production values was found. Microphytobenthos demonstrated significant rates of primary production that is comparable to pelagic production of Kongsfjorden and, hence, emphasised the importance as C source for the zoobenthos.     

Nitrogen and Phosphorous Excretion Rates by Tubificids from the Prahova River (Romania)

Nitrogen and phosphorous exchange at the water–sediment interface is controlled both by complex physico-chemical factors and biological processes. Zoobenthos excretion is one of the most important processes in the mineralization of sedimented organic mater. In polluted freshwaters, tubificid worms are among the dominant components of the benthic community. Rates of ammonium and inorganic phosphate excretion by tubificids were experimentally assessed. They were related to the tubificid abundance in a stream ecosystem polluted with municipal and industrial wastewater. The relationship between these rates and temperature were investigated within the range of 4–23 °C. Relatively constant excretion rates were obtained for both nutrients in the first 8 h of excretion, ranging between 0.076 and 0.226 μg N mg d.w.⁻¹ h⁻¹ and 0.0065–0.01 μg P mg d.w.⁻¹ h⁻¹_, respectively. Q₁₀ values of 2.52 for ammonium and 1.31 for phosphate were calculated. If we presume that all excreta eventually enters the water column, then we can calculate that these invertebrates potentially add 39.17 mg N m⁻² day⁻¹ and 0.49 mg P m⁻² day⁻¹. These values accounts for 17.16 and 7.56% of the nutrient load in the river water, respectively.     

Development of ground vegetation biomass and nutrient pools in a clear-cut disc-plowed boreal forest

Nutrient leaching from forest substrate after clear-cutting and subsequent soil preparation is strongly influenced by the capacity of ground vegetation to sequester the released nutrients. We studied the rates and patterns of biomass and nutrient accumulation in ground vegetation growing on ridges, in furrows and on undisturbed surfaces for 2–5 years after disc-plowing in eastern Finland. The biomass of mosses on ridges remained significantly lower than that in furrows and on undisturbed surfaces. Field layer biomass on ridges and in furrows was significantly lower than on undisturbed surfaces throughout the study period. Field layer biomass increased more on ridges than in furrows. Root biomass on ridges and undisturbed surfaces was considerably higher than in furrows. Five years after disc-plowing, total biomass and nutrient pools for ridges (biomass 4,975 kg ha⁻¹, N 40 kg ha⁻¹, P 5 kg ha⁻¹, K 20 kg ha⁻¹ and Ca 18 kg ha⁻¹) and undisturbed surfaces (biomass 5,613 kg ha⁻¹, N 43 kg ha⁻¹, P 5 kg ha⁻¹, K 22 kg ha⁻¹ and Ca 18 kg ha⁻¹) were similar, but considerably lower for furrows (biomass 1,807 kg ha⁻¹, N 16 kg ha⁻¹, P 2 kg ha⁻¹, K 10 kg ha⁻¹ and Ca 6 kg ha⁻¹). Ridges covered 25% of the area, furrows 30 and 45% was undisturbed surfaces. Taking into account the proportion of each type of surface, values for the whole prepared clear-cut area were 4,312, 34, 4, 18 and 14 kg ha⁻¹ for biomass, N, P, K and Ca, respectively. Biomass and nutrient pools had not returned to uncut forest levels at the end of the 5-year study period. The results indicate that mosses and field layer vegetation respond differently to soil preparation, that the development of biomass on ridges, in furrows and on undisturbed surfaces proceeds at different rates, and that the biomass and nutrient uptake of ground vegetation remains below pre-site preparation levels for several years. However, ridges, which are known to be the most susceptible to leaching, revegetate rapidly. Responsible Editor: Tibor Kalapos.     

Differential growth response of <Emphasis Type="Italic">Ulva lactuca</Emphasis> to ammonium and nitrate assimilation

Controlled cultivation of marine macroalgal biomass such as Ulva species, notably Ulva lactuca, is currently studied for production of biofuels or functional food ingredients. In a eutrophic environment, this macrophyte is exposed to varying types of nutrient supply, including different and fluctuating levels of nitrogen sources. Our understanding of the influences of this varying condition on the uptake and growth responses of U. lactuca is limited. In this present work, we examined the growth response of U. lactuca exposed to different sources of nitrogen (NH₄⁺; NO₃⁻; and the combination NH₄NO₃) by using photo-scanning technology for monitoring the growth kinetics of U. lactuca. The images revealed differential increases of the surface area of U. lactuca disks with time in response to different N-nutrient enrichments. The results showed a favorable growth response to ammonium as the nitrogen source. The NH₄Cl and NaNO₃ rich media (50 μM of N) accelerated U. lactuca growth to a maximum specific growth rate of 16.4 ± 0.18% day⁻¹ and 9.4 ± 0.72% day⁻¹, respectively. The highest biomass production rate obtained was 22.5 ± 0.24 mg DW m⁻²·day⁻¹. The presence of ammonium apparently discriminated the nitrate uptake by U. lactuca when exposed to NH₄NO₃. Apart from showing the significant differential growth response of U. lactuca to different nitrogen sources, the work exhibits the applicability of a photo-scanning approach for acquiring precise quantitative growth data for U. lactuca as exemplified by assessment of the growth response to two different N-sources.     

Kinetics of growth, oxygen uptake, and substrate utilization of wild type and genetically engineeredBurkholderia sp

The gene (vgb) encoding the hemoglobin (VHb) ofVitreoscilla sp. was cloned intoBurkholderia sp. and the effect of VHb on the growth characteristics of genetically engineeredBurkholderia (YV1) were compared with wild typeBurkholderia (R34) using continuous flow reactors (chemostat) at various dilution rates under aerobic conditions. Batch oxygen uptake rate showed that YV1 has much higher oxygen uptake rate than R34 (i.e. 0.63 mg O₂/g biomass/min vs. 1.43 mg O₂/g biomass/min for R34 and YV1 respectively at a dilution rate of 1.2 day⁻¹). Monod parameters, maximum growth rate (μ_max) and half saturation coefficient (K_s) were found to be 7.03 day⁻¹ and 691 mg/L for R34 respectively, compared to 5.49 day⁻¹ and 404 mg/L for YV1 respectively. At low dilution rates (<2.5 day⁻¹), when the substrate is present in low concentrations, the growth yield was much higher in YV1 (0.52) than in R34 (0.37). Although substrate utilization rates were similar between R34 and YV1, the latter showed much higher oxygen uptake rate than did R34 at all dilution rates. When the stability of VHb was tested on agar plates containing 40 μg/L of kanamycin and 100 μg/L of ampicillin,vgb gene containing VHb plasmid in YV1 was stable over 82 days. When survivability under oxygen limited conditions was tested, R34 survived only for 11 days whereas YV1 survived over 25 days in liquid media; in agar plate experiments, R34 did not survive more than 40 days whereas more than 75% of YV1 survived over 110 days.     

Modelling the influence of bioturbation on the vertical distribution of sedimentary phosphorus in L. Esrom

A multilayer sediment-water exchange model was used to evaluate the importance of bioturbation in the profundal sediments of L. Esrom. The temporal variation of the vertical distribution of sedimentary phosphorus fractions was modelled with an objective function of 1.50. Deviations between measured and simulated values occurred in the spring, where the measured pool of sedimentary phosphorus sharply declined in the surface sediments. The application of a model for the activity ofChironomus anthracinus based on biomass, oxygen consumption and temperature improved the model in the spring period. The downwards transport of easy-degradable surface sediments reduced the average release of sedimentary phosphorus from 12 mg P · m⁻² · day⁻¹ to 11 mg P · M⁻² · day⁻¹. The introduction of a similar model for the other important burrowing species in L. Esrom,Potamothrix hammoniensis, lowered the objective function to 1.37 and increased the average release to 12.5 mg P · m⁻² · day⁻¹. The minor role of bioturbation in sediment processes is discussed.