Microbial decomposition of synthetic C-labeled lignins in nature: lignin biodegradation in a variety of natural materials

Lignin biodegradation in a variety of natural materials was examined using specifically labeled synthetic C-lignins. Natural materials included soils, sediments, silage, steer bedding, and rumen contents. Both aerobic and anaerobic incubations were used. No C-labeled lignin biodegradation to labeled gaseous products under anaerobic conditions was observed. Aerobic C-labeled lignin mineralization varied with respect to type of natural material used, site, soil type and horizon, and temperature. The greatest observed degradation occurred in a soil from Yellowstone National Park and amounted to over 42% conversion of total radioactivity to CO(2) during 78 days of incubation. Amounts of C-labeled lignin mineralization in Wisconsin soils and sediments were significantly correlated with organic carbon, organic nitrogen, nitrate nitrogen, exchangeable calcium, and exchangeable potassium.     

Degradation of softwood [14C lignin] lignocelluloses and its relation to the formation of humic substances in river and pond environments

The fate of lignin in water and sediment of the Garonne river (France) and of a pond in its floodplain was examined using specifically labeled [¹⁴C-lignin] lignocelluloses. No significant differences appeared in the mineralization rate of alder, poplar or willow [¹⁴C-lignin] in running water samples. Conversion of total radioactivity to ¹⁴CO₂ ranged between 18.7% and 24.4% after 120 days of incubation. Degree of ¹⁴C-labeled lignin mineralization in standing water and sediments was clearly lower, especially in submerged sediments, and was correlated with oxygen supply. After 60 days of incubation 3.3% to 7.9% of the ¹⁴C-labeled lignin was recovered in water samples as dissolved organic carbon originating from microbial metabolism. In water extracts from sediment the percentage of dissolved organic ¹⁴C was only 0.4% to 1.3% of the applied activity. In the humic fraction extracted from sediments it did not exceed 4.4% which was much lower than in soils. No significant difference appeared between river and pond conditions for humic substances formation.     

Absence of microbial mineralization of lignin in anaerobic enrichment cultures

The existence of anaerobic biodegradation of lignin was examined in mixed microflora. Egyptian soil samples, in which rapid mineralization of organic matter takes place in the presence of an important anaerobic microflora, were used to obtain the anaerobic enrichment cultures for this study. Specifically, 14CO2 or [14C]lignin wood was used to investigate the release of labeled gaseous or soluble degradation products of lignin in microbial cultures. No conversion of 14C-labeled lignin to 14CO2 or 14CH4 was observed after 6 months of incubation at 30 degrees C in anaerobic conditions with or without NO3-. A small increase in soluble radioactivity was observed in certain cultures, but it could not be related to the release of catabolic products during the anaerobic biodegradation of lignin.     

Absence of microbial mineralization of lignin in anaerobic enrichment cultures.

The existence of anaerobic biodegradation of lignin was examined in mixed microflora. Egyptian soil samples, in which rapid mineralization of organic matter takes place in the presence of an important anaerobic microflora, were used to obtain the anaerobic enrichment cultures for this study. Specifically, 14CO2 or [14C]lignin wood was used to investigate the release of labeled gaseous or soluble degradation products of lignin in microbial cultures. No conversion of 14C-labeled lignin to 14CO2 or 14CH4 was observed after 6 months of incubation at 30 degrees C in anaerobic conditions with or without NO3-. A small increase in soluble radioactivity was observed in certain cultures, but it could not be related to the release of catabolic products during the anaerobic biodegradation of lignin.     

Methodological Modifications for Accurate and Efficient Determination of Contaminant Biodegradation in Unsaturated Calcareous Soils

Many techniques for quantifying microbial biodegradation of ¹⁴C-labeled compounds use soil-water slurries and trap mineralization-derived ¹⁴CO₂ in solution wells suspended within the incubation flasks. These methods are not satisfactory for studies of arid-region soils that are highly calcareous and unsaturated because (i) slurries do not simulate unsaturated conditions and (ii) the amount of CO₂ released from calcareous soils exceeds the capacity of the suspended well. This report describes simple, inexpensive methodological modifications for quantifying microbial degradation of [¹⁴C]benzene and 1,2-dichloro[U-¹⁴C]ethane in calcareous soils under unsaturated conditions. Soils at 50% water holding capacity were incubated with labeled contaminants for periods up to 10 weeks, followed by acidification of the soil and trapping of the evolved CO₂ in a separate container of 2 N NaOH. The CO₂ was transferred from the incubation flask to the trap solution by a gas transfer shunt containing activated charcoal to remove any volatilized labeled organics. The amount of ¹⁴CO₂ in the trap solution was measured by scintillation counting (disintegrations per minute). The method was tested by using two regional unamended surface soils, a sandy aridisol and a clay-rich riparian soil. The results demonstrated that both [¹⁴C]benzene and 1,2-dichloro[U-¹⁴C]ethane were mineralized to release substantial amounts of ¹⁴CO₂ within 10 weeks. Levels of mineralization varied with contaminant type, soil type, and aeration status (anaerobic vs. aerobic); no significant degradation was observed in abiotic control samples. Methodological refinements of this technique resulted in total ¹⁴CO₂ recovery efficiency of approximately 90%.     

Soil Ca alters processes contributing to C and N retention in the Oa/A horizon of a northern hardwood forest

Recent studies on the effects of calcium (Ca) additions on soil carbon (C) cycling in organic soil horizons present conflicting results, with some studies showing an increase in soil C storage and others a decrease. We tested the legacy effects of soil Ca additions on C and nitrogen (N) retention in a long-term incubation of soils from a plot-scale field experiment at the Hubbard Brook Experimental Forest, NH, USA. Two levels of Ca (850 and 4250 kg Ca/ha) were surface applied to field plots as the mineral wollastonite (CaSiO₃) in summer of 2006. Two years after field Ca additions, Oa/A horizon soils were collected from field plots and incubated in the laboratory for 343 days to test Ca effects on C mineralization, dissolved organic carbon (DOC) export, and net N transformations. To distinguish mineralization of soil organic C (SOC) from that of more recent C inputs to soil, we incubated soils with and without added ¹³C-labeled sugar maple leaf litter. High Ca additions increased exchangeable Ca and pH compared to the control. While low Ca additions had little effect on mineralization of SOC or added litter C, high Ca additions reduced mineralization of SOC and enhanced mineralization of litter C. In litter-free incubations, δ¹³C of respired C was enriched in the high Ca treatment compared to the control, indicating that Ca suppressed mineralization of ¹³C-depleted SOC sources. Leaching of DOC and NH₄ ⁺ were reduced by Ca additions in litter-free and litter-amended soils. Our results suggest that Ca availability in these organic soils influences mineralization of SOC and N primarily by stabilization processes and only secondarily through pH effects on organic matter solubility, and that SOC binding processes become important only with relatively large alterations of Ca status.     

Lignin decomposition is sustained under fluctuating redox conditions in humid tropical forest soils

Lignin mineralization represents a critical flux in the terrestrial carbon (C) cycle, yet little is known about mechanisms and environmental factors controlling lignin breakdown in mineral soils. Hypoxia is thought to suppress lignin decomposition, yet potential effects of oxygen (O₂) variability in surface soils have not been explored. Here, we tested the impact of redox fluctuations on lignin breakdown in humid tropical forest soils during ten‐week laboratory incubations. We used synthetic lignins labeled with ¹³C in either of two positions (aromatic methoxyl or propyl side chain C_β) to provide highly sensitive and specific measures of lignin mineralization seldom employed in soils. Four‐day redox fluctuations increased the percent contribution of methoxyl C to soil respiration relative to static aerobic conditions, and cumulative methoxyl‐C mineralization was statistically equivalent under static aerobic and fluctuating redox conditions despite lower soil respiration in the latter treatment. Contributions of the less labile lignin C_β to soil respiration were equivalent in the static aerobic and fluctuating redox treatments during periods of O₂ exposure, and tended to decline during periods of O₂ limitation, resulting in lower cumulative C_β mineralization in the fluctuating treatment relative to the static aerobic treatment. However, cumulative mineralization of both the C_β‐ and methoxyl‐labeled lignins nearly doubled in the fluctuating treatment relative to the static aerobic treatment when total lignin mineralization was normalized to total O₂ exposure. Oxygen fluctuations are thought to be suboptimal for canonical lignin‐degrading microorganisms. However, O₂ fluctuations drove substantial Fe reduction and oxidation, and reactive oxygen species generated during abiotic Fe oxidation might explain the elevated contribution of lignin to C mineralization. Iron redox cycling provides a potential mechanism for lignin depletion in soil organic matter. Couplings between soil moisture, redox fluctuations, and lignin breakdown provide a potential link between climate variability and the biochemical composition of soil organic matter.     

Biodegradation of atrazine under denitrifying conditions

Anaerobic biodegradation of atrazine by the bacterial isolate M91-3 was characterized with respect to mineralization, metabolite formation, and denitrification. The ability of the isolate to enhance atrazine biodegradation in anaerobic sediment slurries was also investigated. The organism utilized atrazine as its sole source of carbon and nitrogen under anoxic conditions in fixed-film (glass beads) batch column systems. Results of HPLC and TLC radiochromatography suggested that anaerobic biotransformation of atrazine by microbial isolate M91-3 involved hydroxyatrazine formation. Ring cleavage was demonstrated by ¹⁴CO₂ evolution. Denitrification was confirmed by detection of ¹⁵N₂ in headspace samples of K¹⁵NO₃-amended anaerobic liquid cultures. In aquatic sediments, mineralization of uniformly ring-labeled [¹⁴C]atrazine occurred in both M91-3-inoculated and uninoculated sediment. Inoculation of sediments with M91-3 did not significantly enhance anaerobic mineralization of atrazine as compared to uninoculated sediment, which suggests the presence of indigenous organisms capable of anaerobic atrazine biodegradation. Results of this study suggest that the use of M91-3 in a fixed-film bioreactor may have applications in the anaerobic removal of atrazine and nitrate from aqueous media. Received: 3 September 1997 / Received revision: 4 December 1997 / Accepted: 2 January 1998     

Nitrogen mineralization and its controlling factors in various kinds of temperate humid-zone soils

The N mineralization capacity of 41 temperate humid-zone soils of NW Spain was measured by aerobic incubation for 15 days at 28°C and 75% of field capacity. The main soil factors affecting organic N dynamics were identified by principal components analysis. Ammonification predominated over nitrification in almost all soils. The mean net N mineralization rate was 1.63% of the organic N content, and varied according to soil parent materials as follows: soils on basic and ultrabasic rocks < soils over acid metamorphic rocks < soils developed over sediments < soils over acid igneous rocks < soils on limestone. The N mineralization capacity was lower in natural soils than in cropped soils or pastures. The accumulation of organic matter (C and N) seems to be due to poor mineralization which was caused, in decreasing order of importance, by high exchangeable H-ion levels, high Al and Fe gel contents and, to a lesser extent (though more markedly in cropped soils), by silty clay texture and exchangeable Al ions.     

Thermophilic Anaerobic Biodegradation of [14C]Lignin, [14C]Cellulose, and [14C]Lignocellulose Preparations

Thermophilic (55°C) anaerobic enrichment cultures were incubated with [¹⁴C-lignin]lignocellulose, [¹⁴C-polysaccharide]lignocellulose, and kraft [¹⁴C]lignin prepared from slash pine, Pinus elliottii, and ¹⁴C-labeled preparations of synthetic lignin and purified cellulose. Significant but low percentages (2 to 4%) of synthetic and natural pine lignin were recovered as labeled methane and carbon dioxide during 60-day incubations, whereas much greater percentages (13 to 23%) of kraft lignin were recovered as gaseous end products. Percentages of label recovered from lignin-labeled substrates as dissolved degradation products were approximately equal to percentages recovered as gaseous end products. High-pressure liquid chromatographic analyses of CuO oxidation products of sound and degraded pine lignin indicated that no substantial chemical modifications of the remaining lignin polymer, such as demethoxylation and dearomatization, occurred during biodegradation. The polysaccharide components of pine lignocellulose and purified cellulose were relatively rapidly mineralized to methane and carbon dioxide; 31 to 37% of the pine polysaccharides and 56 to 63% of the purified cellulose were recovered as labeled gaseous end products. An additional 10 to 20% of the polysaccharide substrates was recovered as dissolved degradation products. Overall, these results indicate that elevated temperatures can greatly enhance rates of anaerobic degradation of lignin and lignified substrates to methane and low-molecular-weight aromatic compounds.     

蚂蚁筑巢对西双版纳热带森林土壤有机氮矿化的影响

蚂蚁作为生态系统工程师能够调节土壤微生物及理化环境,进而对热带森林土壤有机氮矿化速率及其时间动态产生显著影响。以西双版纳白背桐热带森林群落为研究对象,采用室内需氧培养法测定土壤有机氮矿化速率,比较蚁巢和非蚁巢土壤有机氮矿化速率的时间动态,揭示蚂蚁筑巢活动引起土壤无机氮库、微生物生物量碳及化学性质改变对有机氮矿化速率时间动态的影响。结果表明：（1）蚂蚁筑巢显著影响土壤有机氮矿化速率（P<0.01）,相较于非蚁巢,蚁巢土壤有机氮矿化速率提高了261%;（2）土壤有机氮矿化速率随月份推移呈明显的单峰型变化趋势,即6月最大（蚁巢1.22 mg kg^-1 d^-1、非蚁巢0.41 mg kg^-1 d^-1）,12月最小（蚁巢0.82 mg kg^-1 d^-1、非蚁巢0.18 mg kg^-1 d^-1）;（3）两因素方差分析表明,不同月份及不同处理对土壤有机氮矿化速率、NH₄-N及NO₃-N产生显著影响（P<0.05）,但对NO₃-N的交互作用不显著;（4）蚂蚁筑巢显著提高了无机氮库（NH₄-N与NO₃-N）、微生物生物量碳、有机质、水解氮、全氮及易氧化有机碳等土壤养分含量,而降低了土壤pH值;（5）回归分析表明,铵态氮和硝态氮对土壤有机氮矿化速率产生显著影响,分别解释87.89%、61.84%的有机氮矿化速率变化;（6）主成份分析表明NH₄-N、微生物生物量碳及有机质是影响有机氮矿化速率时间动态的主要因素,而全氮、NO₃-N、易氧化有机碳、水解氮及pH对土壤有机氮矿化速率的影响次之,且pH与土壤有机氮矿化速率呈显著负相关。总之,蚂蚁筑巢活动主要通过影响土壤NH₄-N、微生物生物量碳及有机质的状况,进而调控西双版纳热带森林土壤有机氮矿化速率的时间动态。研究结果将有助于进一步提高对土壤氮矿化生物调控机制的认识。     

Effect of Mineral and Organic Soil Constituents on Microbial Mineralization of Organic Compounds in a Natural Soil

This research addressed the effect of mineral and organic soil constituents on the fate of organic compounds in soils. Specifically, it sought to determine how the associations between organic chemicals and different soil constituents affect their subsequent biodegradation in soil. Four ¹⁴C-labeled surfactants were aseptically adsorbed to montmorillonite, kaolinite, illite, sand, and humic acids. These complexes were mixed with a woodlot soil, and ¹⁴CO₂ production was measured over time. The mineralization data were fitted to various production models by nonlinear regression, and a mixed (3/2)-order model was found to most accurately describe the mineralization patterns. Different mineralization patterns were observed as a function of the chemical and soil constituents. Surfactants that had been preadsorbed to sand or kaolinite usually showed similar mineralization kinetics to the control treatments, in which the surfactants were added to the soil as an aqueous solution. Surfactants that had been bound to illite or montmorillonite were typically degraded to lesser extents than the other forms, while surfactant-humic acid complexes were degraded more slowly than the other forms. The desorption coefficients (K_d) of the soil constituent-bound surfactants were negatively correlated with the initial rates of degradation (k₁) and estimates of ¹⁴CO₂ yield (P_o) as well as actual total yields of ¹⁴CO₂. However, there was no relationship between K_d and second-stage zero-order rates of mineralization (k_o). Microbial community characteristics (biomass and activity) were not correlated with any of the mineralization kinetic parameters. Overall, this study showed that environmental form had a profound effect on the ultimate fate of biodegradable chemicals in soil. This form is defined by the physicochemical characteristics of the chemical, the composition and mineralogy of the soil, and the mode of entry of the chemical into the soil environment.     

Effect of concentration and environmental form of tetradecenyl succinic acid on its mineralization in soil

Tetradecenyl succinic acid (TSA) is the major component of a detergent builder (C12-C14 alkenyl succinic acid), which is inherently biodegradable. ¹⁴C-TSA was dosed as a component of sewage sludge into a soil with a history of sludge amendment at final added concentrations of 1.5 and 30 mg (kg soil)^-1. In addition, it was dosed to the soil in an aqueous solution to a final added concentration of 30 mg (kg soil)^-1. Dose and form were found to have a pronouced effect on the mineralization kinetics. When dosed in a realistic form and concentration (i.e. 1.5 mg (kg soil)^-1 as a component of sludge), TSA was mineralized at its highest rate and to its greatest extent, and the mineralization half-life was 2.4 days. When dosed at 30 mg (kg soil)^-1 as a component of sludge, mineralization began immediately, and the half-life was 23 days. In contrast, when dosed at this concentration in aqueous solution, the onset of mineralization was preceded by a 13 day lag period and the mineralization half-life was 69 days. Primary biodegradation and mineralization rates of TSA were very similar. Approximately, half the radioactivity was evolved as ¹⁴CO₂, while the remaining radioactivity became non-extractable, having presumably been incorporated into biomass or natural soil organic matter (humics). This study demonstrated that TSA is effectively removed from sludge-amended soils as a result of biodegradation. Furthermore, it showed the effect that dose form and concentration have on the biodegradation kinetics and the importance of dosing a chemical not only at a relevant concentration but also in the environmental form in which it enters the soil environment.     

Lignocellulose Mineralization by Arctic Lake Sediments in Response to Nutrient Manipulation

Mineralization of specifically labeled ¹⁴C-cellulose- and ¹⁴C-lignin-labeled lignocelluloses by Toolik Lake, Alaska, sediments was examined in response to manipulation of various environmental factors. Mineralization was measured by quantifying the amount of labeled CO₂ released from the specifically labeled substrates. Nitrogen (NH₄NO₃) and, to a greater degree, phosphorus (PO₄⁻³) additions enhanced the mineralization of white pine (Pinus strobus) cellulose during the summer of 1978. Nitrogen and phosphorus together had no cumulative effect. During the summer of 1979, nitrogen or phosphorus alone had only a slight stimulatory effect on the mineralization of a sedge (Carex aquatilis) cellulose; however, together, they had a dramatic effect. This variable response of mineralization to nutrient addition between 1978 and 1979 was probably attributable to year-to-year variation in nutrient availability within the lake. Cellobiose addition and oxygen depletion inhibited the amount of pine cellulose mineralized. Whereas addition of nitrogen to oxygen-depleted treatments had limited effect, addition of phosphorus resulted in mineralizations equal to or greater than that of the controls. Nitrogen had no effect on mineralization of pine or Carex lignins. Phosphorus, however, inhibited mineralization of both lignins. With Carex lignin, the phosphorus inhibition occurred at a concentration as low as 0.1 μM. The antagonistic role of phosphorus in cellulose and lignin mineralizations may be of significance in understanding the increased proportion of lignin relative to cellulose in decomposing litter.     

Litter type and soil minerals control temperate forest soil carbon response to climate change

Temperate forest soil organic carbon (C) represents a significant pool of terrestrial C that may be released to the atmosphere as CO₂ with predicted changes in climate. To address potential feedbacks between climate change and terrestrial C turnover, we quantified forest soil C response to litter type and temperature change as a function of soil parent material. We collected soils from three conifer forests dominated by ponderosa pine (PP; Pinus ponderosa Laws.); white fir [WF; Abies concolor (Gord. and Glend.) Lindl.]; and red fir (RF; Abies magnifica A. Murr.) from each of three parent materials, granite (GR), basalt (BS), and andesite (AN) in the Sierra Nevada of California. Field soils were incubated at their mean annual soil temperature (MAST), with addition of native ¹³C‐labeled litter to characterize soil C mineralization under native climate conditions. Further, we incubated WF soils at PP MAST with ¹³C‐labeled PP litter, and RF soils at WF MAST with ¹³C‐labeled WF litter to simulate a migration of MAST and litter type, and associated change in litter quality, up‐elevation in response to predicted climate warming. Results indicated that total CO₂ and percent of CO₂ derived from soil C varied significantly by parent material, following the pattern of GR>BS>AN. Regression analyses indicated interactive control of C mineralization by litter type and soil minerals. Soils with high short‐range‐order (SRO) mineral content exhibited little response to varying litter type, whereas PP litter enriched in acid‐soluble components promoted a substantial increase of extant soil C mineralization in soils of low SRO mineral content. Climate change conditions increased soil C mineralization greater than 200% in WF forest soils. In contrast, little to no change in soil C mineralization was noted for the RF forest soils, suggesting an ecosystem‐specific climate change response. The climate change response varied by parent material, where AN soils exhibited minimal change and GR and BS soils mineralized substantially greater soil C. This study corroborates the varied response in soil C mineralization by parent material and highlights how the soil mineral assemblage and litter type may interact to control conifer forest soil C response to climate change.     

Evaluating Biodegradation as a Primary and Secondary Treatment for Removing RDX (Hexahydro-1,3,5-trinitro-1,3,5-triazine) from a Perched Aquifer

Ground water beneath the U.S. Department of Energy (USDOE) Pantex Plant is contaminated with the high explosive RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine). The authors evaluated biodegradation as a remedial option by measuring RDX mineralization in Pantex aquifer microcosms spiked with ¹⁴C-labeled RDX (75 g soil, 15 ml of 5 mg RDX/L). Under anaerobic conditions and constant temperature (16°C), cumulative ¹⁴CO₂ production ranged between 52% and 70% after 49 days, with nutrient-amended (C, N, P) microcosms yielding the greatest mineralization (70%). The authors also evaluated biodegradation as a secondary treatment for removing RDX degradates following oxidation by permanganate (KMnO₄) or reduction by dithionite-reduced aquifer solids (i.e., redox barriers). Under this coupled abiotic/biotic scenario, we found that although unconsumed permanganate initially inhibited biodegradation, > 48% of the initial ¹⁴C-RDX was recovered as ¹⁴CO₂ within 77 days. Following exposure to dithionite-reduced solids, RDX transformation products were also readily mineralized (> 47% in 98 days). When we seeded Pantex aquifer material into Ottawa Sand that had no prior exposure to RDX, mineralization increased 100%, indicating that the Pantex aquifer may have an adapted microbial community that could be exploited for remediation purposes. These results indicate that biodegradation effectively transformed and mineralized RDX in Pantex aquifer microcosms. Additionally, biodegradation may be an excellent secondary treatment for RDX degradates produced from in situ treatment with permanganate or redox barriers.     

An Investigation of the Ability of a 14C-Labeled Hydrocarbon Mineralization Test to Predict Bioremediation of Soils Contaminated with Petroleum Hydrocarbons

Bioremediation is a widely accepted technology for the remediation of hydrocarbon-contaminated soil. Treatability studies are usually carried out to assess the biodegradation potential of the contaminants and to design optimal treatments. Laboratory studies measuring soil respiration are often used. One method consists of monitoring the mineralization of a ¹⁴C-labeled hydrocarbon surrogate added to the contaminated soil. This study investigates the ability of this method to properly predict the removal of the hydrocarbon contaminants initially found in soils. Mineralization of ¹⁴C-labeled hexadecane was monitored in seven soils contaminated with various hydrocarbon mixtures, both fresh and weathered, in microcosm experiments. Reduction of total petroleum hydrocarbon (TPH) concentrations was measured simultaneously in separate microcosms. Both types of microcosms were subjected to the same amendment regimes. For all soils, poor correlation was observed between the mineralization and TPH reduction data sets. Mineralization data supported contaminants removal data in only one soil. Findings indicate that the radioactive surrogate method does not reliably predict the extent of, and the effect of amendments on, the removal of the hydrocarbons initially present in soil, and may therefore predict suboptimal treatment regimes. Recommendations for soil treatability protocols are provided.     

Effect of Concentration of Organic Chemicals on Their Biodegradation by Natural Microbial Communities

The effect of concentration on the biodegradation of synthetic organic chemicals by natural microbial communities was investigated by adding individual ¹⁴C-labeled organic compounds to stream water at various initial concentrations and measuring the formation of ¹⁴CO₂. The rate of degradation of p-chlorobenzoate and chloroacetate at initial concentrations of 47 pg/ml to 47 μg/ml fell markedly with lower initial concentrations, although half or more of the compound was converted to CO₂ in 8 days or less. On the other hand, little mineralization of 2,4-dichlorophenoxyacetate and 1-naphthyl-N-methylcarbamate, or the naphthol formed from the latter, occurred when these compounds were present at initial concentrations of 2 to 3 ng/ml or less, although 60% or more of the chemical initially present at higher concentrations was converted to CO₂ in 6 days. It is concluded that laboratory tests of biodegradation involving chemical concentrations greater than those in nature may not correctly assess the rate of biodegradation in natural ecosystems and that low substrate concentration may be important in limiting biodegradation in natural waters.     

Decomposition of [14C]Lignocelluloses of Spartina alterniflora and a Comparison with Field Experiments

Decomposition of lignocelluloses from Spartina alterniflora in salt-marsh sediments was measured by using ¹⁴C-labeled compounds. Rates of decomposition were fastest in the first 4 days of incubation and declined later. Lignins labeled in side chains were mineralized slightly faster than uniformly labeled lignins; 12% of the [side chain-¹⁴C]lignin-labeled lignocellulose was mineralized after 816 h of incubation, whereas only 8% of the [U-¹⁴C]lignin-labeled lignocelluloses were degraded during this period. The carbohydrate moiety within the lignocellulose complex was degraded about four times faster than the lignin moiety; after 816 h of incubation, 29 to 37% of the carbohydrate moiety had been mineralized. Changes in concentration of lignin and cellulose in litter of S. alterniflora were followed over 2 years of decay. Cellulose disappeared from litter more rapidly than lignin; 50% of the initial content of cellulose was lost after 130 days, whereas lignin required 330 to 380 days for 50% loss. The slow loss of lignin compared with other litter components resulted in a progressive enrichment of litter in lignin content. The rates of mineralization of [¹⁴C]lignocelluloses in marsh sediments were similar to the rates of lignocellulose decomposition in litter on the marsh.     

The heterogeneous nature of microbial products as shown by solid-state13C CP/MAS NMR spectroscopy

Homoionic Na-, Ca-, and Al-clays were prepared from the <2 m fractions of Georgia kaolinite and Wyoming bentonite and mixed with sand to give artificial soils with 5, and 25% clay. The artificial soils were inoculated with microbes from a natural soil before incubation. Unlabelled and uniformly¹³C-labelled (99.9% atom) glucose were incorporated into the artificial soils to study the effects of clay types, exchangeable cations and clay contents on the mineralization of glucose-carbon and glucose-derived organic materials. Chemical transformation of glucose-carbon upon incorporation into microbial products and metabolites, was followed using solid-state¹³C CP/MAS NMR spectroscopy.There was a significant influence of exchangeable cations on the mineralization of glucose-carbon over a period of 33 days. At 25% clay content, mineralization of glucose-carbon was highest in Ca-soils and lowest in Al-soils. The influence of exchangeable cations on mineralization of glucose-carbon was more pronounced in soils with bentonite clay than those with kaolinite clay. Statistical analysis of data showed no overall effect of clay type on mineralization of glucose-carbon. However, the interactions of clay type with clay content and clay type with clay content and exchangeable cations were highly significant. At 25% clay content, the mineralization of glucose-carbon was significantly lower in Na- and Al-soils with Wyoming bentonite compared with Na- and Al-soils with Georgia kaolinite. For Ca-soils this difference was not significant. Due to the increased osmotic tension induced by the added glucose, mineralization of glucose-carbon was slower in soils with 5% clay than soils with 25% clay.Despite the differences in the chemical and physical characteristics of soils with Ca-, Na- and Al-clays, the chemical composition of organic materials synthesised in these soils were similar in nature. Assuming CP/MAS is quantitative, incorporation of uniformly¹³C-labelled glucose (99.9% atom) in these soils resulted in distribution of carbon in alkyl (24–25%), O-alkyl (56–63%), carbonyl (11–15%) and small amounts of aromatic and olefinic carbon (2–4%). However, as decomposition proceeded, the chemistry of synthesised material showed some changes with time. In the Ca- and Na-soils, the proportions of alkyl and carbonyl carbon decreased and that of O-alkyl carbon increased with time of incubation. However, the opposite trend was found for the Al-soil.Proton-spin relaxation editing (PSRE) subspectra clearly showed heterogeneity within the microbial products. Subspectra of the slowly-relaxing (long T₁(H)) domains were dominated by alkyl carbon in long- and short-chain structures. The signals due to N-alkyl (55 ppm) and carbonyl carbon were also strong in these subspectra. These subspectra were very similar to those obtained for microbial and fungal materials and were probably microbial tissues attached to clay surfaces by polysaccharide extracellular mucilage. Subspectra of fast-relaxing (short T₁(H)) domains comprised mostly O-alkyl and carbonyl carbon and were probably microbial metabolites released as neutral and acidic sugars into the extracellular environment, and strongly sorbed by clay surfaces.