Ammonium regeneration and carbon utilization by marine bacteria grown on mixed substrates

We examined the impact of exposing natural populations of marine bacteria (from seawater collected near Woods Hole, Massachusetts, USA) to multiple nitrogen and carbon sources in a series of batch growth experiments conducted from 1989 through 1990. The substrate C:N ratio (C:N_s) was varied from 1.5:1 to 10:1 either with equal amounts of NH ₄ ⁺ and different amino acids or an amino acid mixture, all supplemented with glucose to maintain the C:N_s ratio equal to that of the respective amino acid, or with combinations of glucose and NH ₄ ⁺ alone. A common feature of the experiments involving amino acids was the concurrent uptake of NH ₄ ⁺ and amino acids that persisted as long as a readily assimilable carbon source (glucose in our case) was taken up. There was no net regeneration of NH ₄ ⁺ , even though catabolism of amino acids occurred. Regeneration of NH ₄ ⁺ was evident only after glucose was completely utilized, which usually occurred at the end of exponential growth. The contribution of¹⁵NH ₄ ⁺ to total nitrogen uptake by the end of exponential growth varied from ~60 to 80% when individual amino acids were present and down to ~24% when the amino acid mixture was added. These estimates are conservative because we did not account for possible isotope dilution effects resulting from amino acid catabolism. When NH ₄ ⁺ and glucose were the sole nitrogen and carbon sources, there was a stoichiometric balance between glucose and NH ₄ ⁺ uptake over a wide range of C:N_s ratios, leading to a constant bacterial biomass C:N ratio (C:N_B) of ~4.5:1. As a result NH ₄ ⁺ usage varied from 50% when the C:N_s ratio was 3.6:1, to 100% when the C:N_s ratio was 10:1. Gross growth efficiency varied from ~60% when NH ₄ ⁺ plus glucose were added alone or with the amino acid mixture, to 47% when the individual amino acids were used in place of the mixture. It is thus evident that actively growing bacteria will act as sinks for nitrogen when a carbon source that can be assimilated easily is available to balance NH ₄ ⁺ uptake, even when amino acids are available and are being co-metabolized.     

Nitrogenous excretion by the sediment-living bivalve Nucula tenuis from the Oslofjord,Norway

The resting rate of ammonia excretion for the sediment living bivalve Nucula tenuis (Montagu) was found to be 38.8 gN mg^-1 dw h^-1×10^-4 in August and November 1985 in the Oslofjord. The excretion rate of experimental individuals was 37% higher when placed in artificial glass bead sediment. The regression between dry weight and excretion was logN excretion=1.338+1.192 log x, where excretion is gN individual^-1 h^-1×10^-4 and log x=mg dry weight.     

Ammonium regeneration and assimilation in eelgrass (Zostera marina) beds

Regeneration and assimilation of ammonium in the water column and in sediments of eelgrass (Zostera marina L.) beds of Izembek Lagoon and Crane Cove, Alaska, USA and Mangoku-Ura, northeastern Japan, were investigated by using a ¹⁵N isotope dilution technique. In the water column of Mangoku-Ura, ammonium was regenerated at a rate of 12 nmol l^-1 h^-1 and assimilated at a rate of 74 nmol l^-1 h^-1. The ammonium regeneration rate in sediments ranged from 2 to 150 nmol g^-1 h^-1, and with one exception, exceeded ammonium assimilation in sediments (0.3 to 77 nmol g^-1 h^-1). The ammonium regeneration in the water column was of little significance for the nitrogen supply to the eelgrass bed ecosystem. Net ammonium production (regeneration minus assimilation) in the sediment of Izembek Laggon met nitrogen demand for eelgrass growth, suggesting that ammonium regeneration in the sediments was very important for the nitrogen cycle in the eelgrass bed ecosystem.     

Urea as a nitrogen source for the phytoplankton in the Oslofjord

Ambient concentrations of urea in the inner Oslofjord, Norway, showed a pronounced yearly cycle in 1980, with values in the range 0.1 to 10.0 μg-at N l^-1; this cycle resemble that of ammonia although urea concentrations were usually lower. The uptake of urea by phytoplankton was investigated using ¹⁵N. Urea was usually a less important N source than NH ₄ ⁺ , and accounted for 0 to 53% (mean 19%) of summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rates from April to October. Absolute as well as relative (specific) uptake rates of urea were higher in the summer (June–August) than at other times. Uptake of urea was inhibited by NH ₄ ⁺ concentrations higher than 1 to 2 μg-at N l^-1. The summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rate was exponentially related to temperature.     

Experimental evidence for internal predation in microzooplankton communities

The fate of microzooplankton production, whether it is channeled to mesozooplankton or recycled within the microbial food web, has major implications for the oceanic carbon cycle. The aim of this study was to estimate internal predation within naturally occurring microzooplankton communities. A dilution series based on the Landry and Hasset technique was created by mixing 200-μm-screened water (used as whole water) with 5-μm-screened seawater due to the dominance of pico- and small nanoplankton at our study site. This modification of the original technique allows for gradual reduction in microzooplankton abundance and thus internal predation while maintaining sufficient phytoplankton prey levels for microzooplankton growth in diluted treatments. Microzooplankton growth and mortality rates were calculated based on the changes in abundance during 24-h incubation. In the diluted treatments, microzooplankton growth rates were significantly higher (1.21 ± 0.20 day^?1 for ciliates and 0.88 ± 0.05 day^?1 for heterotrophic dinoflagellates) compared to those in whole seawater where microzooplankton abundance remained unchanged or even declined over time. Approximately 79 % of microzooplankton production was consumed within the microzooplankton, with aloricate ciliates being the most vulnerable to predation. These findings support the assumption that trophic interactions between microzooplankton can be an important factor controlling their production and, thus, energy transfer in picoplankton-dominated pelagic ecosystems.     

Phytoplankton blooms are strongly impacted by microzooplankton grazing in coastal North Pacific waters

Phytoplankton growth and microzooplankton grazing were measured in two productive coastal regions of the North Pacific: northern Puget Sound and the coastal Gulf of Alaska. Rates of phytoplankton growth (range: 0.09–2.69 day⁻¹) and microzooplankton grazing (range: 0.00–2.10 day⁻¹) varied seasonally, with lowest values in late fall and winter, and highest values in spring and summer. Chlorophyll concentrations also varied widely (0.19–13.65 μg l⁻¹). Large (>8 μm) phytoplankton cells consistently dominated phytoplankton communities under bloom conditions, contributing on average 65% of total chlorophyll biomass when chlorophyll exceeded 2 μg l⁻¹. Microzooplankton grazing was an important loss process affecting phytoplankton, with grazing rates equivalent to nearly two-thirds (64%) of growth rates on average. Both small and large phytoplankton cells were consumed, with the ratio of grazing to growth (g:μ) for the two size classes averaging 0.80 and 0.42, respectively. Perhaps surprisingly, the coupling between microzooplankton grazing and phytoplankton growth was tighter during phytoplankton blooms than during low biomass periods, with g:μ averaging 0.78 during blooms and 0.49 at other times. This tight coupling may be a result of the high potential growth and ingestion rates of protist grazers, some of which feed on bloom-forming diatoms and other large phytoplankton. Large ciliates and Gyrodinium-like dinoflagellates contributed substantially to microzooplankton biomass at diatom bloom stations in the Gulf of Alaska, and microzooplankton biomass overall was strongly correlated with >8 μm chlorophyll concentrations. Because grazing tended to be proportionally greater when phytoplankton biomass was high, the absolute amount of chlorophyll consumed by microzooplankton was often substantial. In nearly two-thirds of the experiments (14/23), more chlorophyll was ingested by microzooplankton than was available for all other biological and physical loss processes combined. Microzooplankton were important intermediaries in the transfer of primary production to higher trophic levels in these coastal marine food webs. Received: 12 November 1999 / Accepted: 4 October 2000     

Seasonal variations in microzooplankton grazing in the region of the Subtropical Convergence

Microzooplankton grazing and community structure were investigated in the region of the Subtropical Convergence (STC) during three cruises of the South African Antarctic Marine Ecosystem Study (SAAMES) in austral summer (January/February 1993; December 1994/January 1995) and winter (June/July 1993). Chlorophyll a concentrations were consistently dominated by the <20 m size fraction during all three cruises, while the contribution of the microphytoplankton (>20 m) to total chlorophyll a concentrations varied considerably between cruises. Microzooplankton communities were numerically dominated by protozoans comprising ciliates (aloricates and tintinnids) and dinoflagellates. Instantaneous growth coefficients of phytoplankton in the vicinity of the STC showed no seasonal trends. However, marked seasonal differences were observed in the size structure of the phytoplankton. The grazing impact of microzooplankton was highest when the <20 m chlorophyll fraction contributed >95% of the total. Under these conditions, the instantaneous grazing rates ranged between 0.15 and 0.66 d^-1. These correspond to daily losses of 14 to 48% of the inntial standing stock and between 45 and 81% of the potential primary production. At stations where microphytoplankton contributed significantly (-20%) to total chlorophyll concentrations, the grazing coefficients were lower, ranging between 0 and 0.53 d^-1. This corresponds to a loss of <41% of the initial standing stock, or between 0 and 56% of the potential production. Our data suggest that microzooplankton represent the main grazing sink for production when the <20 m chlorophyll size-class dominates total chlorophyll. These facts suggest that the efficiency of the biological pump may vary over time.     

The impact of grazing by microzooplankton in northern Hiroshima Bay,the Seto Inland Seam,Japan

The abundance of microzooplankton and their grazing impact on phytoplankton were studied using the dilution technique from May 1990 to November 1991 in northern Hiroshima Bay, a typical eutrophic area in the Seto Inland Sea. Microzooplankton, dominated in number by tintinnid ciliates, were abundant from June to September when chlorophyll-a concentrations were high. Maximum density of microzooplankton ranged from 3.8×10³ to 25.4×10³ ind l^-1. During the period of investigation, mean microzooplankton density and mean chlorophyll-a concentration of the <20-m fraction increased toward the inner region of the bay. The microzooplankton grazing on phytoplankton increased from summer to early autumn, and decreased from late autumn to winter. At an offshore station, the annual means of the daily grazing loss for total chlorophyll-a and the chlorophyll-a of the <20-m fraction were 12 and 15% of the initial standing stock, respectively. At an estuarine station, the microzooplankton grazed 19 and 29% of the total and <20-m initial standing stock, respectively. The quantity of grazed chlorophyll-a correlated positively and linearly with the potential production of chlorophyll-a at both stations. The quantity of chlorophyll-a grazed by microzooplankton and the potential production of chlorophyll-a were nearly equivalent in the <20-m fraction at the estuarine station. This suggests that the microzooplankton assemblage was able to consume almost all the nanoplankton newly produced in the eutrophic estuary.     

Ammonium cycling by Antarctic zooplankton in winter

Elemental composition and excretion rates of ammonium-nitrogen of zooplankton, ranging over more than five orders of magnitude in body size, were measured in mid-winter in coastal waters west of the Antarctic Peninsula. Excretion rates were constant for the initial 12 h of incubation in the four species tested, and experimental stocking densities of up to 126 mg dry wt l^-1 did not cause variability in the rate of ammonium production. Weight-specific excretion rates of freshly caught Euchaeta antarctica, Conchoecia sp., Thysanoessa macrura, Euphausia superba, and early stage copepodites of Metridia gerlachei were not significantly different from those reported in summer. However, adult copepods of M. gerlachei and Calanoides acutus appear to have reduced their nitrogen metabolism during winter. Turnover rates of body nitrogen increased with diminishing size, ranging from <0.5% body N d^-1 for large E. superba to >7% body N d^-1 for CII and CIII copepodites of M. gerlachei. Only the nitrogen turnover rates of C. acutus were sufficiently low as to suggest that it could survive the entire austral winter without feeding. Phytoplankton and bacterioplankton were virtually absent in both the water column and the sea-ice. We conclude that carnivory is the dominant trophic mode of the pelagic zooplankton community in Antarctica during winter. Production of ammonium-nitrogen by the zooplankton community probably accounts for M10% of the total ammonium regenerated prior to the annual spring bloom.     

The clearance rate of microzooplankton as the key element for describing estimated non-linear dilution plots demonstrated by a model

The objective of this study was to determine whether the clearance rate of grazers (as an individual response) was sensitive enough to describe non-linear plots estimated by dilution experiments for measuring the instant grazing rate of microzooplankton. The study was based on an initial analysis of a non-linear feeding pattern based on the food concentration dependence of clearance rate of microzooplankton. In contrast to the traditional assumption of a linear functional response, I assumed that the microzooplankton functional response was non-linear and that the dependence of the clearance rate can be sub-divided into four intervals of food concentration (Sections I–IV) as follows: in Section I clearance rate is zero; Section II is a transitional interval in which the clearance rate increases from zero to a maximum value; in Section III, the clearance rate is maximal and constant, and in Section IV, the clearance rate decreases from its maximum value due to saturated ingestion rate. A set of derived differential equations describes the phytoplankton growth rate in each section, leading to the possibility of comparing predicted non-linear dilution plots with observed non-linear dilution data, using only the specific solutions for Sections III and IV. One should evaluate the quality of fit provided by the non-linear and linear models, rather than uncritically accepting only the linear model for observed non-linear dilution data, using calculated expected non-linear and linear dilution plots as alternative hypotheses. It can be demonstrated that the non-linear model provided a better fit to estimated non-linear dilution data from the Red Sea, Rhode River Estuary (USA) and Kiel Fjord (Germany) than the standard linear model. Published dilution experiments which had a non-linear shape were also selected as illustrative examples to demonstrate the superior fit of the non-linear model.     

Stable macrofauna community structure despite fluctuating food supply in subtidal soft sediments of Oslofjord,Norway

At a locality at 32-m depth in Oslofjord, Norway, temperature varied from 4.8° to 9.2°C and salinity varied from 31.2 to 33.3 S over a two-year period. There was a peak in chlorophyll a, and C and N in April–June and a smaller peak in November in the sediment. Bacterial numbers showed maxima in July–August and November–December. The macrobenthic fauna was typical of a species-rich and undisturbed boreal community of silt-clay sediments. The community was predominantly composed of surface and subsurface deposit feeders. Over the twoyear period there was little variation in numerical abundance or biomass of the species despite the variation in food input. The lack of seasonality shown by the fauna probably relates to the lack of variability of the physical environment. The mechanism by which this control is achieved, however, is not known. There are large predators/disturbers in the community such as the polychaetes Lumbrineris fragilis, Glycera rouxii, G. alba, Nephthys spp. and the echinoid Brissopsis lyrifera, which probably play an important role in structuring the community.     

Warming induces shifts in microzooplankton phenology and reduces time-lags between phytoplankton and protozoan production

Indoor mesocosm experiments were conducted to test for potential climate change effects on the spring succession of Baltic Sea plankton. Two different temperature (Δ0?°C and Δ6?°C) and three light scenarios (62, 57 and 49?% of the natural surface light intensity on sunny days), mimicking increasing cloudiness as predicted for warmer winters in the Baltic Sea region, were simulated. By combining experimental and modeling approaches, we were able to test for a potential dietary mismatch between phytoplankton and zooplankton. Two general predator–prey models, one representing the community as a tri-trophic food chain and one as a 5-guild food web were applied to test for the consequences of different temperature sensitivities of heterotrophic components of the plankton. During the experiments, we observed reduced time-lags between the peaks of phytoplankton and protozoan biomass in response to warming. Microzooplankton peak biomass was reached by 2.5 day °C^?1 earlier and occurred almost synchronously with biomass peaks of phytoplankton in the warm mesocosms (Δ6?°C). The peak magnitudes of microzooplankton biomass remained unaffected by temperature, and growth rates of microzooplankton were higher at Δ6?°C (μ_?0?°C?=?0.12 day^?1 and μ_?6?°C?=?0.25 day^?1). Furthermore, warming induced a shift in microzooplankton phenology leading to a faster species turnover and a shorter window of microzooplankton occurrence. Moderate differences in the light levels had no significant effect on the time-lags between autotrophic and heterotrophic biomass and on the timing, biomass maxima and growth rate of microzooplankton biomass. Both models predicted reduced time-lags between the biomass peaks of phytoplankton and its predators (both microzooplankton and copepods) with warming. The reduction of time-lags increased with increasing Q₁₀ values of copepods and protozoans in the tritrophic food chain. Indirect trophic effects modified this pattern in the 5-guild food web. Our study shows that instead of a mismatch, warming might lead to a stronger match between protist grazers and their prey altering in turn the transfer of matter and energy toward higher trophic levels.     

Ammonium excretion by the ophiuridOphiothrix fragilis as a function of season and tide

Ammonium excretion of a dense population (~1 500 individuals m^–2) of the ophiuridOphiothrix fragilis (Abildgaard) was measured in the Dover Straits (French coast) between May 1989 and March 1990: the excretion rate varied from 4.8 µg N g^–1 dry wt h^–1 in November to 12.8 µg N g^–1 dry wt h^–1 in June. Mean individual ammonium excretion,E, wasE=0.019t +1.26 (whereE=µg N individual^–1 andt=time in min;r=0.80;N=81). Variations in the ammonium excretion rate during a tidal cycle appeared to arise from variations in the duration of the suspension-feeding activity ofO. fragilis, which was governed by the strength of the tidal current. During short-term starvation, excretion was low (E=0.009t+1.47;r=0.91;N=17), increasing with increasing length of starvation [E=4.62lnt–2.5;r=0.95;N=17], as observed for other echinoderms; this could be due to catabolism of tissue. The daily ammonia flux from thisO. fragilis population to the water column was estimated at 41 mg N m^–2 d^–1.     

Nitrogen regeneration by the surf zone penaeid prawn Macropetasma africanus

Nitrogen excretion of individual Macropetasma africanus (Balss) from an exposed beach/surf zone in Algoa Bay, South Africa was monitored under laboratory and field conditions in relation to body mass, temperature and feeding during 1985. Excretion rate experiments were performed on starved prawns at 15°, 18°, 20° and 25°C, as well as on individuals fed on four different diets (mussel, fish, shrimp and natural diet) at 15° and 20°C. The ratios of the excreted compounds to total nitrogen excreted were similar for the four diets despite differences in their nitrogen content and in the amount of food consumed. At 15° and 20°C, ammonia excretion rates of fed individuals were four to seven times higher than in starved prawns. the excretion rates were not correlated with nitrogen content of diets. M. africanus recycles 1 557 g NH₄–N per metre strip per year or 1 832 g total nitrogen m^-1 yr^-1, which constitute 12 and 14%, respectively, of total phytoplankton requirements of the surf zone. This study indicates that large motile crustaceans, when abundant, can play an important role in nutrient recycling in turbulent marine environments.     

硅酸钙处理富营养化湖水中氨氮的实验研究

湖泊的富营养化问题日益严重,已给人类的生产、生活带来了极大的危害.而氨氮废水的大量排放,使原本恶化的湖泊水雪上加霜.为了去除富营养化湖水中氨氮,研究了硅酸钙对氨氮的吸附性能.经检测可知水样中氨氮的质量浓度为1.12 mg?L-1,实验结果表明,用粒径大小为100目的硅酸钙吸附剂处理100 mL的水样,当投加量为1.0 g,pH为8,震荡时间为60 min时吸附达到平衡,硅酸钙对富营养化湖水中氨氮的去除率达到81.67%;其吸附等温线符合Langmuir和Freundlich吸附等温式,线性相关系数分别为0.9639和0.9793,最大吸附量为6.60 mg?g-1;由此可见硅酸钙能够很好地吸附富营养化水体中的氨氮.     

Patterns of microzooplankton growth in dilution experiments across a trophic gradient: Implications for herbivory studies

To investigate the growth and grazing patterns of microzooplankton (MZP) in environments of differing productivity, dilution experiments measuring phytoplankton growth (μ) and grazing mortality (m) rates were performed using samples from contrasting locations along the Texas coast. Samples were collected from estuaries, coastal lagoons and offshore Gulf of Mexico locations in the spring and summer of 2001. MZP growth rates were determined in each dilution treatment. Although MZP biomass changed over time in most dilution treatments, adjusting μ and m for the actual grazer gradient (represented by geometric mean MZP biomass) did not cause a significant deviation from the nominal dilution gradient. Likewise, these adjustments did not yield significant regressions where none existed before adjustment. The dynamics of MZP taxonomic groups (ciliates, dinoflagellates) and size categories differed suggesting that in some cases internal predation may lead to trophic cascades. MZP biomass was higher in productive coastal waters and included a larger proportion of dinoflagellates than in the oligotrophic, ciliate-dominated waters of the Gulf of Mexico. The MZP biomass-to-chlorophyll a ratio was lowest in the hypereutrophic Nueces River, where MZP biomass significantly increased in all dilution treatments (net growth rates up to 2 day⁻¹) suggesting a strong top–down control. In the brown-tide dominated Upper Laguna Madre and the oligotrophic seagrass-dominated Lower Laguna Madre MZP growth was decoupled from that of phytoplankton. At these sites, MZP were likely fueled by bacterial carbon and mixotrophy, respectively. Observing the growth response of MZP in dilution experiments can provide insight into trophic structure and efficiency of the microbial food web.     

Ammonium excretion by gelationous zooplankton and their contribution to the ammonium requirements of microplankton in Chesapeake Bay

Ammonium excretion rates of recently collected specimens of gelatinous zooplankton, the scyphomedusan Chrysaora quinquecirrha DeSor and the etenophore Mnemiopsis leidyi A. Agassiz, were correlated with body mass and water temperature in measurements made from April to October 1989 and 1990. Rates ranged between 3.5 and 5.0 g atoms NH ₄ ⁺ -N (g dry wt)^-1h^-1 for C. quinquecirrha and 3.0 to 4.9 g atoms NH ₄ ⁺ -N (g dry wt)^-1h^-1 for M. leidyi. Excretion rate equations and in situ data on the size distributions and biomasses of gelatinous zooplankters and water temperature were used to estimate the contribution of ammonium by medusae and ctenophores to mesohaline Chesapeake Bay waters on several dates during April to October 1989 and 1990. We then compared the estimated contributions to direct measurements of ¹⁵NH ₄ ⁺ uptake by microplankton. The maximum estimated regeneration by gelatinous zooplankton was 5.8 g atoms NH ₄ ⁺ -N m^-3h^-1 at night in August 1990, when medusae biomass was greatest. This represents about 4% of the ammonium required by the microplankton. During the daytime on all dates, less than 1% of the ammonium required by microplanktion was supplied by gelatinous zooplankton. Therefore, gelatinous zooplankton appear to play a minor role in the ammonium cycle of Chesapeake Bay.