Occurrence and Bacterial Cycling of d Amino Acid Isomers in an Estuarine Environment

Abundance of d isomers of amino acids has been used in studies of organic matter diagenesis to determine the contribution of bacterial biomass to the organic matter, especially in marine sediments. However, fluxes of d amino acids in pelagic waters are poorly known. Here we present seasonal changes (March–September) in concentrations of dominant d amino acids in the pool of dissolved free and combined (hydrolysable) amino acids (DFAA and DCAA) in the shallow Roskilde Fjord, Denmark. The amino acid dynamics are related to pelagic bacterial density and activity and abundance of viruses. d␣isomers made up 3.6 and 7.9% of the DFAA and DCAA (average values), respectively, and had similar seasonal variations in concentrations. In batch cultures (0.7- and 0.2-m filtered water in a 1:9 mixture) microbial activity reduced l+d DCAA concentrations in seven of ten sampling dates, while DCAA were released at the remaining three sampling times. NH₄⁺ balance (uptake or release) in the cultures correlated significantly with variations in concentrations of d-DCAA, but not with the total DCAA pools. Abundance of viruses did not correlate with density or production of bacteria in the fjord, but covaried with mineralization of total C, DCAA and PO₄³⁻ in the batch cultures. The content of d amino acids in bacterial biomass in the cultures varied from 6.7 to 12.5% and correlated with the d isomer concentration in the fjord, except for d-Ala. In an additional six-day batch culture study, DCAA and d-DCAA were assimilated by the bacteria during the initial 36 h, but were released between 36 and 42 h simultaneous with a decline in the bacterial density. Our results demonstrate that peptidoglycan components contribute to natural amino acid pools and are assimilated by bacterial assemblages. This cell wall “cannibalism” ensures an efficient recycling of nutrients within the microbial community. Significant positive correlations between viral abundance and bacterial mineralization of organic matter in the fjord indicated that viral lysis contributed to this nutrient recycling.     

Regulation of bacterial proteolytic activity at natural concentrations in dissolved organic matter in a maritime pond]

The regulation of the bacterial exoproteolytic activity, at natural substrate concentrations, was studied during the survey of an Atlantic coastal marine pond (France). The regulation of this activity occurs at two different levels: on the one hand, at the cellular level, the ectoenzyme synthesis is regulated by hydrolysis substrates, dissolved combined amino acids (DCAA), and end products, dissolved free amino acids (DFAA), in terms of the relative amounts available to the cell, and on the other hand, at the ecosystem level, i.e. the hydrolytic activity, by the total amounts of DCAA and DFAA in situ. The DFAA acts as an inhibitor in enzymatic synthesis; in contrast, dissolved proteins induce the enzymatic synthesis and the exoproteolytic activity. These results, obtained in natural concentration conditions, confirm the functioning in situ of the ectoenzymatic activity regulation model of Chróst, until now only validated in an enriched experimental medium.     

Colloidal and dissolved organic matter in lake water: Carbohydrate and amino acid composition,and ability to support bacterial growth

Bacterial utilization of dissolved organic matter (DOM) was studied in water from a humic and a clearwater oligotrophic lake. Indigenous bacteria were inoculated into either 0.2 m natural filtered lake water, or lake water enriched fivefold with colloidal DOM >100 kD but below 0.2 m. Consumption of DOM was followed from changes in concentrations of total dissolved organic carbon (DOC), dissolved combined and free carbohydrates and amino acids (DCCHO and DFCHO, and DCAA and DFAA, respectively) and by uptake of monosaccharide and amino acid radioisotopes. DCCHO and DCAA made up 8% (humic lake) to 33–44% (clear-water lake) of the natural DOC pools, while DFCHO and DFAA contributed at most 1.7% to the DOC pools. Addition of >100 kD DOM increased the DOC concentrations by 50% (clearwater lake) to 92% (humic lake), but it only resulted in a higher bacterial production (by 63%) in the humic lake. During the incubations 13 to 37% of the DOC was assimilated by the bacteria, at estimated growth efficiencies of 4–8%. Despite the measured reduction of DOC, statistically significant changes of specific organic compounds, especially of DCCHO and DCAA, generally did not occur. Probably the presence of high molecular weight DOC interfered with the applied analytical procedures. Addition of radiotracers indicated, however, that DFAA sustained 17–58% and 29–100% of the bacterial carbon and nitrogen requirements, respectively, and that glucose met 1–3% of the bacterial carbon requirements. Thus, our experiments indicate that radiotracers, rather than measurements of concentration changes, should be used in studies of bacterial utilization of DOC in freshwaters with a high content of humic or high molecular weight organic matter.     

Fluxes of free amino acids in three Danish lakes

SUMMARY. 1. Heterotrophic assimilation rates and concentrations of dissolved free amino acids (DFAA) were followed during diel studies in the eutrophic Lake Mossø, Lake Esrom and Lake øm in spring and summer in 1982. In all three lakes, three to four fold diel variations in concentrations and assimilation rates were measured. These fluctuations appeared to be iindependent of phytoplankton and bacteria production. Pools of DFAA varied from 380 nM (Lake Mossø) to 2430 nM (Lake ørn), with serine, glycine, alanine and ornithine as dominant free amino acids.
2. When similar water samples were incubated in a natural light-dark cycle or in total darkness, different pools of DFAA were measured in light and dark.
3. Decomposition of organic matter or zooplankton activity (rather than e.g. phytoplankton exudates) appear to be responsible for the concentration changes.
4. Observed discrepancies between simultaneous concentration changes and assimilation rates are discussed in relation to the applied tracer procedure and the concentration measurements.
5. Assimilation of DFAA sustained from 6% to 25% of the bacterial carbon requirement, corresponding to 2–12% of the phytoplankton production in the lakes.     

Utilization of Dissolved Nitrogen by Heterotrophic Bacterioplankton: Effect of Substrate C/N Ratio

The significance of dissolved combined amino acids (DCAA), dissolved free amino acids (DFAA), and dissolved DNA (D-DNA) as sources of C and N for marine bacteria in batch cultures with variable substrate C/N ratios was studied. Glucose, ammonium, alanine, and phosphate were added to the cultures to produce C/N ratios of 5, 10, and 15 and to ensure that phosphorus was not limiting. Maximum bacterial particulate organic carbon production (after 25 h of incubation) was inversely correlated with the C/N ratio: with the addition of identical amounts of carbon, the levels of production were 9.0-, 10.0-, and 11.1-fold higher at C/N ratios of 15, 10, and 5, respectively, relative to an unamended control. The bacterial growth efficiency increased from 22% (control cultures) to 44 to 53% in the cultures with manipulated C/N ratios (C/N-manipulated cultures). Net carbon incorporation from DCAA, DFAA, and D-DNA supported on average 19, 4, and 3% (control cultures and cultures to which only phosphate was added [+P cultures]) and 5, 4, and 0.3% of the particulate organic carbon production (C/N-manipulated cultures), respectively. In the C/N-manipulated cultures, a 2.6- to 3.4-fold-higher level of incorporation of DCAA, relative to that in the control cultures, occurred. Incorporation of D-DNA increased with the substrate C/N ratio, suggesting that D-DNA mainly was a source of N to the bacteria. Organic N (DCAA, DFAA, and D-DNA) sustained 14 to 49% of the net bacterial N production. NH₄⁺ was the dominant N source and constituted 55 to 99% of the total N uptake. NO₃^- contributed up to 23% to the total N uptake but was released in two cultures. The studied N compounds sustained nearly all of the bacterial N demand. Our results show that the C/N ratio of dissolved organic matter available to bacteria has a significant influence on the incorporation of individual compounds like DCAA and D-DNA.     

Diel variation in concentration,assimilation and respiration of dissolved free amino acids in relation to planktonic primary and secondary production in two eutrophic lakes

Concentration of dissolved free amino acids (DFAA) and assimilation of the 5 most abundant DFAA (glutamic acid, serine, glycine, alanine and ornithine) were measured at 3-h intervals over 27 h in two Danish, eutrophic lakes. The carbon flux of the amino acid assimilation was compared with the major routes of carbon flux, including primary production, bacterial production and zooplankton grazing. In Frederiksborg Slotssø, the mean DFAA concentration was 275 nM with distinct peaks (up to 783 nM) 3 h after sunrise. Assimilation rates of the 5 amino acids amounted on the average to 2.03 µg Cl^–1 h^–1, but high values up to 7.41 µg Cl^–1 h^–1 occurred 3 h after sunrise and at midnight. The mean turnover time of the amino acid pools was 3.2 h. In Lake Mossø, the mean DFAA concentration was 592 nM with peak of 1 161 nM at dusk. The assimilation rate averaged 0.44 µg Cl^–1 h^–1, and the mean turnover time of the amino acid pools was 39 h. In Lake Mossø, similar turnover times of glutamic acid and serine were determined from the ¹⁴C-amino acid tracer technique and Michaelis-Menten uptake kinetics, indicating that the tracer technique gave reliable values of the actual assimilation. The average respiration percentages of the assimilated amino acids were 45% in Frederiksborg Slotssø and 51% in Lake Mossø. Extracellular organic carbon (EOC) released from the phytoplankton contributed DFAA to the water. In Lake Mossø, 81% of the ambient EOC pool was <700 daltons and 9.3% of the EOC was DFAA. This corresponded to about 2.4% of the DFAA pool. Bacterial productivity, determined by means of frequency of dividing cells and ³⁵S-SO₄ dark uptake techniques gave similar results and constituted 4.5 and 3.7 µg Cl^–1 h^–1 in Frederiksborg Slotssø and Lake Mossø, respectively. The bacterial productivity suggested that DFAA were essential substrates to the bacteria, especially in Frederiksborg Slotssø. The zooplankton biomass in Frederiksborg Slotssø was six times larger than that in Lake Mossø, but cladocerans were dominant in both lakes. The zooplankton grazing probably was an important regulatory factor for the bacterial productivity.     

Utilization of Dissolved Nitrogen by Heterotrophic Bacterioplankton: a Comparison of Three Ecosystems

The contributions of different organic and inorganic nitrogen and organic carbon sources to heterotrophic bacterioplankton in batch cultures of oceanic, estuarine, and eutrophic riverine environments were compared. The importance of the studied compounds was surprisingly similar among the three ecosystems. Dissolved combined amino acids (DCAA) were most significant, sustaining from 10 to 45% of the bacterial carbon demands and from 42 to 112% of the bacterial nitrogen demands. Dissolved free amino acids (DFAA) supplied 2 to 7% of the carbon and 6 to 24% of the nitrogen incorporated into the bacterial biomass, while dissolved DNA (D-DNA) sustained less than 5 and 12% of the carbon and nitrogen requirements, respectively. Ammonium was the second most important source of nitrogen, meeting from 13 to 45% of the bacterial demand in the oceanic and estuarine cultures and up to 270% of the demand in riverine cultures. Nitrate was taken up in the oceanic cultures (uptake equaled up to 46% of the nitrogen demand) but was released in the two others. Assimilation of DCAA, DFAA, and D-DNA combined supplied 43% of the carbon demand of the bacteria in the oceanic cultures, while approximately 25% of the carbon requirements were met by the three substrates at the two other sites. Assimilation of nitrogen from DCAA, DFAA, D-DNA, NH₄⁺, and NO₃^-, on the other hand, exceeded production of particulate organic nitrogen in one culture at 27 h and in all cultures over the entire incubation period (50 h). These results suggest that the studied nutrient sources may fully support the nitrogen needs but only partially support the carbon needs of microbial communities of geographically different ecosystems. Furthermore, a comparison of the initial concentrations of the different substrates indicated that relative pool sizes of the substrates seemed to influence which substrates were primarily being utilized by the bacteria.     

Are dissolved free amino acids free?

Microbial assimilation of 3 amino acids (glutamic acid, alanine, and ornithine) was characterized in 3 lakes and 2 marine stations using the Michaelis-Menten kinetic approach. The calculated K_t + S_n concentrations were related to chemical concentration measurements of dissolved free amino acids (DFAA) to evaluate the biological and the chemical determinations of the DFAA pools. Concentrations of K_t + S_n always were larger than chemical measurements of the S_n concentrations. K_t + S_n and S_n varied from 11.5 and 9.5 nM (alanine, oligotrophic lake) to 288.7 and 89.9 nM (ornithine, marine harbor station), respectively. Subtracting S_n from the K_t + S_n concentrations, K_t was found to range from 12–897% of the chemically measured S_n concentrations. To test whether the DFAA actually were free, dissolved molecules, dissolved material in the water samples was separated into various molecular size classes by means of gel permeation chromatography. From 47–116% of the DFAA in the untreated water samples was recovered in the low molecular fraction (<700 Daltons). Variation in recoveries mainly appeared to be due to an incomplete chromatographic separation and difficulties in obtaining proper blank levels. The present observations suggest that labeled tracers can be used in the study of DFAA assimilation and that the DFAA are free, dissolved molecules. This partly conflicts with previously published reports.     

Net release of dissolved organic matter by the scleractinian coral Acropora pulchra

The net production of dissolved organic matter (DOM) and dissolved combined and free amino acids (DCAA and DFAA, respectively) by the hermatypic coral Acropora pulchra was measured in the submerged condition, and the production rates were normalized to the coral surface area, tissue biomass, and net photosynthetic rates by zooxanthellae. When normalized to the unit surface area, the production rates of dissolved organic carbon and nitrogen (DOC and DON, respectively) were 37 and 4.4 nmol cm^− 2 h^− 1, respectively. Comparing with the photosynthetic rate by zooxanthellae, which was measured by ¹³C-tracer accumulation in the soft tissue of the coral colony, the release rate of DOC corresponded to 5.4% of the daily net photosynthetic production. The tissue biomass of the coral colony was 178 µmol C cm^− 2 and 23 µmol N cm^− 2, indicating that the release of DOC and DON accounted for 0.021% h^− 1 and 0.019% h^− 1 of the tissue C and N, respectively. The C:N ratios of the released DOM (average 8.4) were not significantly different from those of the soft tissue of the coral colonies (average 7.7). While DFAA did almost not accumulate in the incubated seawater, DCAA was considerably released by the coral colonies at the rate of 2.1 nmol cm^− 2 h^− 1 on average. Calculating C and N contents of the hydrolyzable DCAA, it was revealed that about 20% and 50%–60% of the released bulk DOC and DON, respectively, were composed of DCAA.     

Diel Production and Microheterotrophic Utilization of Dissolved Free Amino Acids in Waters Off Southern California

Diel patterns of dissolved free amino acid (DFAA) concentration and microheterotrophic utilization were examined in the spring and fall of 1981 in euphotic waters from the base of the mixed layer off the southern California coast. The average depths of the isotherms sampled were 19.2 m for spring and 9.0 m for fall. Total DFAA levels were generally higher in the spring than in the fall, 18 to 66 nM and 14 to 20 nM, respectively. Two daily concentration maxima and minima were observed for total DFAAs as well as for most individual DFAAs. Maxima were usually measured in the mid-dark period and in the early afternoon; minima were typically observed in early morning and late afternoon. Bacterial cell numbers reached maximal values near midnight in both seasons. These increases coincided with one of the total DFAA maxima. The second total DFAA maximum occurred in early to midafternoon, during the time of maximum photosynthetic carbon production and rapid dissolved amino acid utilization. Microbial metabolism (incorporation plus respiration) of selected ³H-amino acids was 2.7 to 4.1 times greater during the daylight hours. DFAA turnover times, based on these metabolic measurements, ranged between 11 and 36 h for the amino acids tested, and rates were 1.7 to 3.7 times faster in the daylight hours than at night. DFAA distributions were related to primary production and chlorophyll a concentrations. Amino acids were estimated to represent 9 to 45% of the total phytoplankton exudate. Microheterotrophic utilization or production of total protein amino acids was estimated as 3.6 μg of C liter⁻¹ day⁻¹ in spring and 1.9 μg of C liter⁻¹ day⁻¹ in the fall. Assimilation efficiency for dissolved amino acids averaged 65% for marine microheterotrophs.     

Bacterial Production in the Recently Flooded Sep Reservoir: Diel Changes in Relation to Dissolved Carbohydrates and Combined Amino Acids

The spatial distribution of bacterial abundance and production were measured every 4 h in a recently flooded oligo-mesotrophic reservoir (the Sep Reservoir, Puy-De-Dôme, France), in relation to concentrations of dissolved carbohydrates and combined amino acids. The concentration of dissolved organic matter (DOM) components in the recently flooded Sep Reservoir were higher than those measured in other lakes of similar trophic status. Short-term variations in the bacterial production in this new reservoir appeared cyclical and endogenous to bacterial communities. These results highlight the need for the evaluation of diel changes in bacterial production, if estimation of the daily production rate of bacteria is to be done accurately for a reliable model of carbon flow through bacterioplankton and ultimately through aquatic microbial food webs. Bacterial growth, measured over time and space, did not appear exclusively governed by DOM components from phytoplankton primary production.     

Bacterial Consumption of Humic and Non-Humic Low and High Molecular Weight DOM and the Effect of Solar Irradiation on the Turnover of Labile DOM in the Southern Ocean

The decomposition of dissolved organic matter (DOM) in pelagic ecosystems is mediated primarily by heterotrophic bacteria, but transformation by short-wave solar radiation may play an important role in surface waters, in particular when humic substances constitute a substantial fraction of the DOM pool. Most of the studies examining bacterial decomposition and photochemical transformation of DOM stem from limnetic and coastal marine systems and much less information is available from oceanic environments. To examine the bacterial decomposition of humic and non-humic DOM in the Southern Ocean we carried out microcosm experiments in which we measured bacterial growth on isolated fractions of humic and non-humic DOM of the size classes <3 kDa and >3 kDa. Experiments carried out at the Polar Front showed a preferential bacterial growth on non-humic DOM and in particular on the size fraction <3 kDa. Bacterial growth, measured as bacterial biomass production, on non-humic DOM accounted for 74% to 88% of the total growth on all four DOM fractions. In experiments in the Antarctic circumpolar current and the coastal current under pack ice, bacterial growth was 6× lower than at the Polar Front, and humic and non-humic DOM was consumed to equal amounts. The size fraction <3 kDa was always preferred. Experiments examining the effect of solar radiation on the release of dissolved amino acids (DAA) and carbohydrates (DCHO) and their subsequent bacterial utilization showed a stimulating effect on glucose uptake and the release of DAA at the Polar Front but an inhibition in the eastern Weddell Sea. Ultraviolet-B was the most effective component of the solar radiation spectrum tested. Effects of UV-B on glucose uptake and release of DAA were positively correlated with concentrations of humic-bound DAA. The data imply that at low concentrations, e.g., <100 nM (amino acid equivalent), UV-irradiation reduces, whereas at concentrations >100 nM UV-irradiation stimulates glucose uptake and release of DAA as compared to dark conditions.     

Concentrations and fluxes of organic carbon substrates in the aquatic environment

Data concerning concentrations and fluxes of dissolved organic compounds (DOC) from marine and lacustrine environments are reviewed and discussed. Dissolved free amino acids and carbohydrates comprised the main fraction in the labile organic carbon pool. Dissolved free amino acids in marine waters varied between 3–1400 nM and those of freshwaters between 2.6–4124 nM. Dissolved free carbohydrates varied between 0.4–5000 nM in marine systems and between 14–1111 nM in freshwaters. The turnover times of both substrate pools varied in marine waters between 1.4 hours and 948 days and in freshwaters between 2 hours and 51 days. Measurements of stable^12/13C-ratio and¹⁴C-isotope dating in ocean deep water samples revealed DOC turnover times between 2000–6000 years. Studies on carbon flows within the aquatic food webs revealed that about 50% of photosynthetically fixed carbon was channelled via DOC to the bacterioplankton. Excreted organic carbon varied between 1–70% of photosynthetically fixed carbon in marine waters and between 1–99% in freshwaters. The labile organic carbon pool represented only 10–30% of the DOC. The majority (70–90%) of the DOC was recalcitrant to microbial assimilation. Only 10–20% of the DOC could be easily chemically identified. Most of the large bulk material represented dissolved humic matter and neither the chemical structure nor the ecological function of the DOC is as yet clearly understood.Abbreviations ATP Adenosine Tri-Phosphate - AMS Accelerated Mass Spectrometry - BSA Bovine Serum Albumin - GlAse GlucosidAse activity - DAA Dissolved Amino Acids - DCAA Dissolved Combined Amino Acids - DFAA Dissolved Free Amino Acids - DTAA Dissolved Total Amino Acids - DCHO Dissolved Carbohydrates - DCCHO Dissolved Combined Carbohydrates - DFCHO Dissolved Free Carbohydrates - DTCHO Dissolved Total Carbohydrates - DLCFaAc Dissolved Long Chain Fatty Acids - DSCFaAc Dissolved Short Chain Fatty Acids - DOC Dissolved Organic Carbon - DOM Dissolved Organic Matter - DHM Dissolved Humic Matter - DTPhOH Dissolved Total Phenolic compounds - DCPhOH Dissolved Combined Phenolic compounds - DFPhOH Dissolved Free Phenolic conpounds - EOC Excreted Organic Carbon - HS Humic Substances - HPLC High Performance Liquid Chromatography - HTCO High-Temperature Catalytic Oxidation - (K_t+S_n) Transport Constant + Natural Substrate from Michaelis Menten Kinetics - LOCP Labile Organic Carbon Pool - OM Organic Matter - MEE Microbial Extracellular Enzymes - PER Percent of Extracellular Release - PhDOC Photosynthetically derived Dissolved Organic Carbon - POC Particulate Organic Carbon - ROCP Recalcitrant Organic Carbon Pool - T_t Turnover time - UDOC Utilizable Dissolved Organic Carbon - V_max Maximum Uptake Velocity - WCO Wet Chemical Oxidation Dedicated to Prof. Drs. J. Overbeck on the occasion of his 70th birthday     

Zooplankton induced changes in dissolved free amino acids and in production rates of freshwater bacteria

This study examined the importance of zooplankton in the flux of dissolved free amino acids (DFAA) in the water and into bacteria. DFAA release rates were followed in laboratory grazing experiments usingDaphnia galeata andEudiaptomus graciloides as grazers, andScenedesmus acutus andSynechococcus elongatus as food sources. Except for minor initial peaks, DFAAs were released continuously during the first 2 hours and made up 6–12% (in one experiment 50%) of the calculated ingestion rates. During three diel studies in lakes, effects of removal and increase of the density of zooplankton (>200m) on the pools of DFAA as well as on the bacterial production were followed. During two of the diel studies, higher DFAA pools were measured when 3–4 times the natural zooplankton density was present, and in one study a minor increase also occurred in the bacterial production, compared with results from experiments without zooplankton and with a natural zooplankton density. The increase in bacterial growth coincided with a decline in DFAA. During the third study, neither DFAA nor the bacterial production changed significantly when the zooplankton density was increased 3 times. Removal of zooplankton, however, caused a decline in both DFAA and bacterial production. Our data suggest a close relationship between occurrence of zooplankton and release of DFAA, but the factors regulating the amount of DFAA released and its effect on bacterial growth are not yet understood.     

Seasonal changes in the dissolved free amino acid and DOC concentrations in a hypertrophic African reservoir and its inflowing rivers

Dissolved free amino acid (DFAA) concentration and composition and dissolved organic carbon (DOC) concentration were measured over 16 months at three depths in hypertrophic Hartbeespoort Dam, South Africa and in its two perenially inflowing rivers. The range of DFAA concentrations in the reservoir and both rivers were similar with dominant DFAA consisting of serine, glycine, alanine and ornithine in all three systems. The range of DOC concentrations in the rivers was 1.5–11.1 mg l^–1, the major river (Crocodile) having about twice the DOC concentration of the Magalies River. The DFAA/DOC ratios ranged between 0.02–1.1% in the Crocodile River and 0.13–3.7% in the Magalies River. DFAA and DOC concentrations were positively correlated to the Magalies River flow, but for the Crocodile River, which received domestic and industrial effluents, DOC was inversely correlated to flow. The source of DFAA in both rivers was mainly terrestrial, in contrast to the main DOC source in the Crocodile River which was the effluents. The DFAA load of the Crocodile River ranged between 0.22 and 208 kg C d^–1.DOC (5.0–24.8mg l^–1) in Hartbeespoort Dam generally decreased with depth but DFAA (15–4800 nmol l^–1) concentration showed no clear trend. The DFAA/DOC ratios varied between 0.02 and 2.9%. DFAA concentrations were correlated (r = 0.3, n = 30, p = 0.04) with bacterial numbers at 0 and 10 m only while no significant correlations were found with bacterial production, chlorophyll a concentration and phytoplankton primary and EDOC (extracellular DOC) production at any depth. The rate of bacterial utilization of DFAA was low compared with data from other lakes. Diurnal phytoplankton production of DFAA in the euphotic zone of the whole lake was calculated to vary between 268 and 30 780 t C d^–1 indicating autochthonous DFAA sources were dominant to allochthonous DFAA sources. The autochthonous production of DFAA was > 2 × gross bacterial production of the euphotic zone indicating that although DFAA concentrations were frequently < 10 g C l^–1, the rate of DFAA production exceeded bacterial requirements.     

Bacterial Contribution to Dissolved Organic Matter in Eutrophic Lake Kasumigaura,Japan

Incubation experiments using filtered waters from Lake Kasumigaura were conducted to examine bacterial contribution to a dissolved organic carbon (DOC) pool. Bacterial abundance, bacterial production, concentrations of DOC, total dissolved amino acids (TDAA), and total dissolved neutral sugars (TDNS) were monitored during the experiments. Bacterial production during the first few days was very high (20 to 35 μg C liter⁻¹ day⁻¹), accounting for 40 to 70% of primary production. The total bacterial production accounted for 34 to 55% of the DOC loss during the experiment, indicating high bacterial activities in Lake Kasumigaura. The DOC degradation was only 12 to 15%, whereas the degradation of TDAA and TDNS ranged from 30 to 50%, suggesting the preferential usage of TDAA and TDNS. The contribution of bacterially derived carbon to a DOC pool in Lake Kasumigaura was estimated using d-amino acids as bacterial biomarkers and accounted for 30 to 50% of the lake DOC. These values were much higher than those estimated for the open ocean (20 to 30%). The ratio of bacterially derived carbon to bulk carbon increased slightly with time, suggesting that the bacterially derived carbon is more resistant to microbial degradation than bulk carbon. This is the first study to estimate the bacterial contribution to a DOC pool in freshwater environments. These results indicate that bacteria play even more important roles in carbon cycles in freshwater environments than in open oceans and also suggests that recent increases in recalcitrant DOC in various lakes could be attributed to bacterially derived carbon. The potential differences in bacterial contributions to dissolved organic matter (DOM) between freshwater and marine environments are discussed.     

Growth response of four freshwater algal species to dissolved organic nitrogen of different concentration and complexity

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Contrasting effects of solar UV radiation on dissolved organic sources for bacterial growth

The photochemical transformation of dissolved organic matter (DOM) in lakes and oceans has been shown to either reduce or enhance bacterial utilization. We compared the effects of UV radiation on the bacterial use of DOM in a wide range of lakes. Although complex DOM was converted in all irradiated samples into carboxylic acids that are readily utilized by bacteria, irradiation in several lakes resulted in a decreased ability of DOM to support bacterial growth. The effect of irradiation on the ability of DOM to promote bacterial growth was a positive function of the terrestrial humic matter, and a negative function of indigenous algal production. We suggest that the net effect of irradiation is a result of counteracting but concurrent processes rendering DOM either labile or recalcitrant. Humic DOM is predominantly transformed into forms of increased lability, whereas photochemical transformation into compounds of decreased bacterial substrate quality dominates in algal-derived DOM. Hence, solar-induced photochemical reactions interact with microbial degraders in different ways, depending on the origin and nature of the organic matter, affecting the transfer of energy within aquatic food webs, as well as the degradation and preservation of detrital organic matter, in different directions.     

Incorporation of [h]leucine and [h]valine into protein of freshwater bacteria: field applications

Incorporation of leucine and valine into proteins of freshwater bacteria as a measure of bacterial production was tested in two eutrophic Danish lakes and was related to bacterial production measured by thymidine incorporation. In a depth profile (0 to 8 m) in Frederiksborg Castle Lake, incorporation of 100 nM leucine and valine gave similar rates of protein production. In terms of carbon, this production was about 50% lower than incorporation of 10 nM thymidine. In another depth profile in the same lake, incorporations of 10 nM valine and 100 nM leucine were identical, but differed from incorporations of 10 nM leucine and 100 nM valine. Bacterial carbon production calculated from incorporations of 10 nM thymidine and 10 nM leucine was similar, whereas 10 nM valine and 100 nM leucine and valine indicated an up to 2.4-fold-higher rate of carbon production. In a diel study in Lake Bagsvaerd, incorporation of 100 nM leucine and valine indicated a similar protein production, but the calculated carbon production was about 1.9-fold higher than the production based on uptake of 10 nM thymidine. Different diel changes in incorporation of the two amino acids and in incorporation of thymidine were observed. In both lakes, concentrations of naturally occurring leucine and valine were <5 nM in most samples. This means that the specific activity of a H isotope added at a concentration of 100 nM usually was diluted a maximum of 5%. Net assimilation of natural free amino acids in the lakes sustained 8 to 69% of the net bacterial carbon requirement, estimated from incorporation of leucine, valine, or thymidine. The present results indicate that incorporation of leucine and valine permits realistic measurements of bacterial production in freshwater environments.