Bacterial Degradation of Dissolved Organic Matter from Two Northern Michigan Streams

This study used high-pressure size exclusion chromatography (HPSEC) to measure the changes in molecular weight distributions of dissolved organic matter (DOM) of two Northern Michigan streams following inoculation with bacterial concentrates from the same locations. During the initial 12 h of the experiment, weight average molecular weight (M _w ) of DOM decreased, as high molecular weight components were lost from solution. After 12 h, the M _w of DOM increased, primarily because of a loss of intermediate to lower molecular weight components. Leucine incorporation showed little or no bacterial metabolism during the first 12 h, but metabolism increased substantially after 12 h. The initial loss of high molecular weight components during the period of little or no bacterial metabolism suggests preferential adsorption of these components to the bacterial surfaces, perhaps followed by metabolism. This suggested interpretation is consistent with previous observations of preferential adsorption of higher molecular weight components to viable but non-metabolizing Bacillus subtilis and to mineral surfaces. The latter loss of lower molecular weight components was most likely due to bacterial metabolism of the DOM, which is consistent with previous observations that lower molecular weight components are more biodegradable. The HPSEC technique uses 254 nm wavelength for detection and focuses primarily on humic- and fulvic-type components rather than low molecular weight organic molecules, such as carbohydrates. Thus, results confirmed that humic/fulvic components are biodegradable, but did not address other DOM components.     

Dynamics and Characterization of Refractory Dissolved Organic Matter Produced by a Pure Bacterial Culture in an Experimental Predator-Prey System

We studied the effects of a bacterium (Pseudomonas chlororaphis) and a bactivorous protozoan (Uronema sp.) on transformations of labile dissolved organic carbon (DOC). In 36-day time series experiments, bacteria were grown on glucose both with and without protozoa. We measured bulk organic carbon pools and used electrospray ionization mass spectrometry to characterize dissolved organic matter on a molecular level. Bacteria rapidly utilized glucose, depleting it to nondetectable levels and producing new DOC compounds of higher molecular weight within 2 days. Some of these new compounds, representing 3 to 5% of the initial glucose-C, were refractory and persisted for over a month. Other new compounds were produced and subsequently used by bacteria during the lag and exponential growth phases, pointing to a dynamic cycling of organic compounds. Grazers caused a temporary spike in the DOC concentration consisting of labile compounds subsequently utilized by the bacteria. Grazing did not increase the complexity of the DOC pool already established by the bacteria but did continually decrease the particulate organic carbon pool and expedited the conversion of glucose-C to CO₂. After 36 days, 29% of initial glucose-C remained in pure bacteria cultures, while only 6% remained in cultures where a grazer was present. In this study the bacteria were the primary shapers of the complex DOC continuum, suggesting higher trophic levels possibly have less of an impact on the qualitative composition of DOC than previously assumed.     

Composition,Dynamics, and Fate of Leached Dissolved Organic Matter in Terrestrial Ecosystems: Results from a Decomposition Experiment

Fluxes of dissolved organic matter (DOM) are an important vector for the movement of carbon (C) and nutrients both within and between ecosystems. However, although DOM fluxes from throughfall and through litterfall can be large, little is known about the fate of DOM leached from plant canopies, or from the litter layer into the soil horizon. In this study, our objectives were to determine the importance of plant-litter leachate as a vehicle for DOM movement, and to track DOM decomposition [including dissolve organic carbon (DOC) and dissolved organic nitrogen (DON) fractions], as well as DOM chemical and isotopic dynamics, during a long-term laboratory incubation experiment using fresh leaves and litter from several ecosystem types. The water-extractable fraction of organic C was high for all five plant species, as was the biodegradable fraction; in most cases, more than 70% of the initial DOM was decomposed in the first 10 days of the experiment. The chemical composition of the DOM changed as decomposition proceeded, with humic (hydrophobic) fractions becoming relatively more abundant than nonhumic (hydrophilic) fractions over time. However, in spite of proportional changes in humic and nonhumic fractions over time, our data suggest that both fractions are readily decomposed in the absence of physicochemical reactions with soil surfaces. Our data also showed no changes in the ¹³C signature of DOM during decomposition, suggesting that isotopic fractionation during DOM uptake is not a significant process. These results suggest that soil microorganisms preferentially decompose more labile organic molecules in the DOM pool, which also tend to be isotopically heavier than more recalcitrant DOM fractions. We believe that the interaction between DOM decomposition dynamics and soil sorption processes contribute to the ¹³C enrichment of soil organic matter commonly observed with depth in soil profiles. published online 2004     

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.     

Limitation of Bacterial Growth by Dissolved Organic Matter and Iron in the Southern Ocean

The importance of resource limitation in controlling bacterial growth in the high-nutrient, low-chlorophyll (HNLC) region of the Southern Ocean was experimentally determined during February and March 1998. Organic- and inorganic-nutrient enrichment experiments were performed between 42°S and 55°S along 141°E. Bacterial abundance, mean cell volume, and [³H]thymidine and [³H]leucine incorporation were measured during 4- to 5-day incubations. Bacterial biomass, production, and rates of growth all responded to organic enrichments in three of the four experiments. These results indicate that bacterial growth was constrained primarily by the availability of dissolved organic matter. Bacterial growth in the subtropical front, subantarctic zone, and subantarctic front responded most favorably to additions of dissolved free amino acids or glucose plus ammonium. Bacterial growth in these regions may be limited by input of both organic matter and reduced nitrogen. Unlike similar experimental results in other HNLC regions (subarctic and equatorial Pacific), growth stimulation of bacteria in the Southern Ocean resulted in significant biomass accumulation, apparently by stimulating bacterial growth in excess of removal processes. Bacterial growth was relatively unchanged by additions of iron alone; however, additions of glucose plus iron resulted in substantial increases in rates of bacterial growth and biomass accumulation. These results imply that bacterial growth efficiency and nitrogen utilization may be partly constrained by iron availability in the HNLC Southern Ocean.     

Production of Urea by Bacterial Decomposition of Organic Matter Including Phytoplankton

A concentrated colony of Fragilaria crotonensis collected from the surface water of Lake Suwa, which is one of the typical eutrophic lakes in Japan, and organic matter contained in untreated surface water from the same lake were subjected to aerobic decomposition by bacteria in a dark room at a temperature of 20 ± 3 °C. An exponential increase of urea with time was recorded in both of the experiments. The apparent rate constants of urea production were calculated to be 0.083 day⁻¹ for decomposition of F. crotonensis and 0.051 day⁻¹ for decomposition of the organic matter contained in the untreated surface water. This study suggests that urea production by bacterial decomposition of organic matter, including phytoplankton, may be an important source of urea in natural waters under certain conditions.     

Dynamics of Organic Matter Decomposition and Microflora Composition of Forest Litter in Artificial Biogeocenoses

The dynamics of litter stock, microbial biomass, and composition and structure of microbial communities, were studied in the course of soil organic matter transformation during vegetation season. The dynamics of litter stock in coniferous and deciduous forests proved to correlate with the biomass and total abundance of microorganisms, particularly, with the proportion of microfungi in the microbial community.     

Bacterial--Invertebrate Interactions in Uptake of Dissolved Organic Matter

SYNOPSIS. AS compared to integumentary uptake systems of soft-bodiedmarine invertebrates, bacterial systems, in terms of transportconstants, are much better adapted to the low concentrationsof dissolved organic nutrients encountered in coastal and offshorewaters. Bacteria respond to the presence of suitable dissolvedorganic substrates with induction, uptake and multiplication,maintaining the concentrations of dissolved organic matter (DOM)permanently low. At realistic in situ concentrations, epidermaluptake by pelagic and epibenthic animals proceeds at such lowrates that scarcely a substantial proportion of their metabolicneeds is provided by absorption. In marine sediments, wherethe life processes of bacteria and animals are closely interrelated,the macrofauna is sheltered by shells, firm tubes and burrows,which are irrigated by means of overlying water of the watercolumn. Hence, interstitial water with its sometimes higherconcentrations of DOM is scarcely available to sediment-dwellinglarger metazoans. The meiofauna mainly inhabits the few millimetersof the upper sediment layers and the thin halos surroundingirrigated macrofaunal burrows, where sufficient oxygen is available.Unless the magnitude of horizontal water movement, the amountsof diffusional nutrient supply and the percentages, by whichnutrient concentrations are reduced by meiofaunal uptake, areknown, estimations of nutritional benefits from uptake of DOMby meiofauna cannot be made. For all infaunal taxa, bacteriaappear to represent a major food supply.     

Effects of Dissolved Organic Matter Photoproducts and Mineral Nutrient Supply on Bacterial Growth in Mediterranean Inland Waters

Sunlight reacts with dissolved organic matter (DOM) modifying its availability as bacterial substrate. We assessed the impact of DOM photoproducts and mineral nutrient supply on bacterial growth in seven inland waters from the South of Spain, where DOM is characterized by low chromophoric content and long residence time. Factorial experiments were performed with presence vs absence of DOM photoproducts and mineral nutrient supply. In six of the seven experiments, we found a significant and negative effect of DOM photoproducts on bacterial growth and a significant and positive effect of mineral nutrient supply. The interaction of these two factors leaded to a compensation of negative effects of photoproducts by availability of mineral nutrients. Dissolved organic matter diagenetic status and the ionic environment where organic carbon is dissolved can be influencing bacterial DOM processing.     

Temperature Dependence of Photodegradation of Dissolved Organic Matter to Dissolved Inorganic Carbon and Particulate Organic Carbon

Photochemical transformation of dissolved organic matter (DOM) has been studied for more than two decades. Usually, laboratory or “in-situ” experiments are used to determine photodegradation variables. A common problem with these experiments is that the photodegradation experiments are done at higher than ambient temperature. Five laboratory experiments were done to determine the effect of temperature on photochemical degradation of DOM. Experimental results showed strong dependence of photodegradation on temperature. Mathematical modeling of processes revealed that two different pathways engaged in photochemical transformation of DOM to dissolved inorganic carbon (DIC) strongly depend on temperature. Direct oxidation of DOM to DIC dominated at low temperatures while conversion of DOM to intermediate particulate organic carbon (POC) prior to oxidation to DIC dominated at high temperatures. It is necessary to consider this strong dependence when the results of laboratory experiments are interpreted in regard to natural processes. Photodegradation experiments done at higher than ambient temperature will necessitate correction of rate constants.     

Environmental Dissolved Organic Matter Governs Biofilm Formation and Subsequent Linuron Degradation Activity of a Linuron-Degrading Bacterial Consortium

It was examined whether biofilm growth on dissolved organic matter (DOM) of a three-species consortium whose members synergistically degrade the phenylurea herbicide linuron affected the consortium''s integrity and subsequent linuron-degrading functionality. Citrate as a model DOM and three environmental DOM (eDOM) formulations of different quality were used. Biofilms developed with all DOM formulations, and the three species were retained in the biofilm. However, biofilm biomass, species composition, architecture, and colocalization of member strains depended on DOM and its biodegradability. To assess the linuron-degrading functionality, biofilms were subsequently irrigated with linuron at 10 mg liter⁻¹ or 100 μg liter⁻¹. Instant linuron degradation, the time needed to attain maximal linuron degradation, and hence the total amount of linuron removed depended on both the DOM used for growth and the linuron concentration. At 10 mg liter⁻¹, the final linuron degradation efficiency was as high as previously observed without DOM except for biofilms fed with humic acids which did not degrade linuron. At 100 μg liter⁻¹ linuron, DOM-grown biofilms degraded linuron less efficiently than biofilms receiving 10 mg liter⁻¹ linuron. The amount of linuron removed was more correlated with biofilm species composition than with biomass or structure. Based on visual observations, colocalization of consortium members in biofilms after the DOM feed appears essential for instant linuron-degrading activity and might explain the differences in overall linuron degradation. The data show that DOM quality determines biofilm structure and composition of the pesticide-degrading consortium in periods with DOM as the main carbon source and can affect subsequent pesticide-degrading activity, especially at micropollutant concentrations.     

Phosphorus Enhances Uptake of Dissolved Organic Matter in Boreal Streams

Retention of carbon (C), either by physical mechanisms or microbial uptake, is a key driver of the transformation and storage of C and nutrients within ecosystems. Both the molecular composition and nutrient content of organic matter influence the rate at which it is retained in streams, but the relative influence of these characteristics remains unclear. We estimated the effects of nutrient content and molecular composition of dissolved organic C (DOC) on uptake in boreal streams by measuring rates of C retention, in situ, following introduction of leachates derived from alder, poplar, and spruce trees subject to long-term fertilization with nitrogen (N) or phosphorus (P). Leachate C:N varied approximately twofold, and C:P varied nearly 20-fold across species and nutrient treatments. Uptake of DOC was greatest for leachates derived from trees that had been fertilized with P, a finding consistent with P-limitation of uptake and/or preferential sorption of P-containing molecules. Optical measures indicated that leachates derived from the three tree species varied in molecular composition, but uptake of DOC did not differ across species, suggesting weak constraints on retention imposed by molecular composition relative to nutrient limitation. Observed coupling between P and C cycles highlights the potential for increased P availability to enhance DOC retention in headwater streams.     

Effect of Organic Matter Decomposition Level on Bacterial Species Diversity and Composition in Relationship to Pythium Damping-Off Severity

Rhizosphere bacteria were isolated from root tip segments of cucumber seedlings grown in a suppressive, slightly decomposed light-colored peat mix, a conducive, more decomposed dark-colored peat mix, and a suppressive dark peat mix amended with composted hardwood bark. The bacteria were identified by a gas chromatographic fatty acid methyl ester analysis. The total number of taxa recovered from a single root tip segment ranged from 9 to 18. No single taxon predominated on all root tip segments harvested from any of the mixes. The highest relative population density reached by a given taxon on any root tip segment was 45%. Hill's first and second diversity numbers, the modified Hill's ratio, and Hurlbert's rarefaction method, which were used as measures of species diversity, indicated that the organic matter decomposition level of the potting mixes did not affect bacterial species diversity. Bray-Curtis polar ordination and Dice resemblance functions, however, indicated that the organic matter decomposition level of a mix significantly influenced the composition of bacterial species in the rhizosphere. Pseudomonas spp. and other taxa capable of inducing suppression of pythium damping-off predominated in the suppressive mixes. These organisms were absent from the conducive mix, in which Arthrobacter and Bacillus spp. predominated. Although effective bacterial biocontrol agents were isolated from both the suppressive mixes and the conducive mix, the majority were isolated from the less decomposed suppressive mixes. Finally, the efficacy of strains was significantly greater in the slightly decomposed light peat mix than in the decomposed dark peat mix. Natural disease suppression within these mixes was associated with the organic matter decomposition level and the bacterial species compositions of the mixes.     

Hydroxyl Radical Production from Irradiated Arctic Dissolved Organic Matter

The hydroxyl radical (OH·) plays an important role in the environmental chemistry and biogeochemistry of surface waters. OH· acts as a strong oxidant within the irradiated water column, and affects the bioavailability, cycling, and mineralization of dissolved organic matter (DOM), the speciation and redox state of important trace metals e.g., iron and copper, and the fate of persistent organic pollutants (POPs). The generation of this species from irradiated DOM may be especially important in Arctic surface waters during the boreal summer, which contains high levels of DOM and experiences continual solar irradiance. Here, we investigate the OH· produced from laser irradiated Arctic DOM isolated from Toolik Lake, AK (68°38′ N, 149°43′ W). We measured the wavelength dependence of OH· production for aqueous solutions of DOM and report that the greatest OH· production occurs at wavelengths less than 360 nm. OH· production rates ranged from 1.7 (±0.1)×10⁻⁷ M h⁻¹ to 6.4 (±0.2)×10⁻⁷ M h⁻¹, with the rate depending on both irradiation wavelength and to a lesser degree the method used to isolate the DOM matrix. These findings lead to a better understanding of the potentially important photo-oxidation processes that may impact DOM cycling in the Arctic.     

Chemical Characteristics and Origin of Dissolved Organic Matter in the Yukon River

Monthly (or bi-weekly) water samples were collected from the Yukon River, one of the largest rivers in North America, at a station near the US Geological Survey Stevens Village hydrological station, Alaska from May to September 2002, to examine the quantity and quality of dissolved organic matter (DOM) and its seasonal variations. DOM was further size fractionated into high molecular weight (HMW or colloidal, 1 kDa–0.45 μm) and low molecular weight (LMW, <1 kDa) fractions. Dissolved organic carbon (DOC), colored dissolved organic matter (C-DOM) and total dissolved carbohydrate (TCHO) species were measured in the size fractionated DOM samples. Concentrations of DOC were as high as 2830 μmol-C l⁻¹ during the spring breakup in May and decreased significantly to 508–558 μmol-C l⁻¹ during open-water season (June–September). Within the DOC pool, up to 85% was in the colloidal fraction (1 kDa–0.45 μm) in early May. As DOC concentration decreased, this colloidal portion remained high (70–85% of the bulk DOC) throughout the sampling season. Concentrations of TCHO, including monosaccharides (MCHO) and polysaccharides (PCHO), varied from 722 μmol-C l⁻¹ in May to 129 μmol-C l⁻¹ in September, which comprised a fairly constant portion of bulk DOC (24±2%). Within the TCHO pool, the MCHO/TCHO ratio consistently increased from May to September. The C-DOM/DOM ratio and the size fractionated DOM increased from May to September, indicating that DOM draining into the Yukon River contained increased amounts of humified materials, likely related to a greater soil leaching efficiency in summer. The average composition of DOM was 76% pedogenic humic matter and 24% aquagenic CHO. Characteristics of soil-derived humic substances and low chlorophyll-a concentrations support a dominance of terrestrial DOM in Yukon River waters.     

Distinct Optical Chemistry of Dissolved Organic Matter in Urban Pond Ecosystems

Urbanization has the potential to dramatically alter the biogeochemistry of receiving freshwater ecosystems. We examined the optical chemistry of dissolved organic matter (DOM) in forty-five urban ponds across southern Ontario, Canada to examine whether optical characteristics in these relatively new ecosystems are distinct from other freshwater systems. Dissolved organic carbon (DOC) concentrations ranged from 2 to 16 mg C L^-1 across the ponds with an average value of 5.3 mg C L^-1. Excitation-emission matrix (EEM) spectroscopy and parallel factor analysis (PARAFAC) modelling showed urban pond DOM to be characterized by microbial-like and, less importantly, by terrestrial derived humic-like components. The relatively transparent, non-humic DOM in urban ponds was more similar to that found in open water, lake ecosystems than to rivers or wetlands. After irradiation equivalent to 1.7 days of natural solar radiation, DOC concentrations, on average, decreased by 38% and UV absorbance decreased by 25%. Irradiation decreased the relative abundances of terrestrial humic-like components and increased protein-like aspects of the DOM pool. These findings suggest that high internal production and/or prolonged exposure to sunlight exerts a distinct and significant influence on the chemistry of urban pond DOM, which likely reduces its chemical similarity with upstream sources. These properties of urban pond DOM may alter its biogeochemical role in these relatively novel aquatic ecosystems.     

Stormflow Dynamics of Dissolved Organic Carbon and Total Dissolved Nitrogen in a Small Urban Watershed

We examined patterns of dissolved organic carbon (DOC) and total dissolved nitrogen (TDN) loading to a small urban stream during baseflow and stormflow. We hypothesized that lower DOC and TDN contributions from impervious surfaces would dilute natural hydrologic flowpath (i.e., riparian) contributions during storm events in an urban watershed, resulting in lower concentrations of DOC and TDN during storms. We tested these hypotheses in a small urban watershed in Portland, Oregon, over a 3-month period during the spring of 2003. We compared baseflow and stormflow chemistry using Mann–Whitney tests (significant at p<0.05). We also applied a mass balance to the stream to compare the relative significance of impervious surface contributions versus riparian contributions of DOC and TDN. Results showed a significant increase in stream DOC concentrations during stormflows (median baseflow DOC = 2.00 mg l⁻¹ vs. median stormflow DOC = 3.46 mg l⁻¹). TDN streamwater concentrations, however, significantly decreased with stormflow (median baseflow TDN = 0.75 mg l⁻¹ vs. median stormflow TDN = 0.56 mg l⁻¹). During storms, remnant riparian areas contributed 70–74% of DOC export and 38–35% of TDN export to the stream. The observed pattern of increased DOC concentrations during stormflows in this urban watershed was similar to patterns found in previous studies of forested watersheds. Results for TDN indicated that there were relatively high baseflow nitrogen concentrations in the lower watershed that may have partially masked the remnant riparian signal during stormflows. Remnant riparian areas were a major source of DOC and TDN to the stream during storms. These results suggest the importance of preserving near-stream riparian areas in cities to maintain ambient carbon and nitrogen source contributions to urban streams.