Assessing the Impact of Effluents from a Modern Wastewater Treatment Plant on Breakdown of Coarse Particulate Organic Matter and Benthic Macroinvertebrates in a Lowland River

Wastewater treatment plants (WWTP) with insufficient technologies for wastewater purification often cause a distinct nutrient pollution in the receiving streams. The increased concentrations of dissolved nutrients can severely disturb the ecological integrity of streams, which has been recently shown for basic ecosystem processes like mineralization of coarse particulate organic matter (CPOM). The present study investigated the impact of a modern WWTP (Zentralkläranlage Jena) on breakdown rates of CPOM exposed in net bags (1 mm mesh size) to the effluent of a large municipal WWTP and an upstream control site in the Saale River (Thuringia, Germany) from April to October 2005. Control and effluent site differed significantly in water chemistry with increased concentrations of dissolved organic carbon (DOC), ammonium, sulfate, and chloride at the effluent site, while the control site displayed higher concentrations of nitrate. However, breakdown rates of toothpickers and small twigs were not significantly different between the sites, whereas breakdown rate of leaf litter was significantly higher at the effluent site (k?=?0.0124 day^?1) than at the control site (k?=?0.0095 day^?1). Benthic invertebrate assemblages inhabiting the sandy stream bed at both sites were dominated by Chironomidae and Oligochaeta, typical inhabitants of fine sediments. Although the Shannon diversity of the benthic invertebrates was slightly higher at the effluent site (0.85) than at the control site (0.63), no significant difference could be detected. Bacterial numbers in water samples and surface biofilms on glass slides also displayed no significant differences between the two sites. This study showed that the effluents of a WWTP with modern technologies for wastewater purification did not directly affect breakdown rates of CPOM, bacteria numbers in epibenthic biofilms and the water column, and the community composition of sediment inhabiting aquatic macroinvertebrates in an effluent-receiving river with already increased concentrations of dissolved nutrients.     

Operational Optimisation of Pilot Scale Biological Nutrient Removal at the Ciudad Real (Spain) Domestic Wastewater Treatment Plant

The aim of this work is optimising operating conditions for a possibleimplementation of a Biological Nutrient Removal (BNR) process in the Wastewater Treatment Plant (WWTP) of Ciudad Real (Spain). Several factors (hydraulic retention times, anaerobic nitrate concentration, sludge age and wastewater biodegradability) were tested using a pilot scale VIP (Virginia Initiative Plant) activated sludge process and domestic wastewater from the full scale plant. Hydraulic retention times used did not cause changes in N and P removal. P removal was adversely affected by anaerobic NO₃ ^- and improved with higher BOD₅/COD ratios in wastewater. Influence of sludge age was very low in P removal, but N removal was mainly affected by this factor. Final operating conditions were selected taking into account their effects over one of both nutrients. COD and SS removal were always successful. N removal was also easily reached and the main difficulty was P removal. P sludge content was very low (2.5–4%) approximately and was also affected by the same factors tested. The main factor to improve P removal was supposed to be the organic wastewater composition. Wastewater characteristics were modified by using different sources from the WWTP. Volatile fatty acids (VFA) addition to the wastewater by using supernatant of the anaerobic sludge digesters seemed to be the best practical solution for a future BNR implementation in the WWTP.     

Reduction of bacterial growth by a vesicular-arbuscular mycorrhizal fungus in the rhizosphere of cucumber (Cucumis sativus L.)

Summary Cucumber was grown in a partially sterilized sand-soil mixture with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum or left uninoculated. Fresh soil extract was places in polyvinyl chloride tubes without propagules of mycorrhizal fungi. Root tips and root segments with adhering soil, bulk soil, and soil from unplanted tubes were sampled after 4 weeks. Samples were labelled with [³H]-thymidine and bacteria in different size classes were measured after staining by acridine orange. The presence of VAM decreased the rate of bacterial DNA synthesis, decreased the bacterial biomass, and changed the spatial pattern of bacterial growth compared to non-mycorrhizal cucumbers. The [³H]-thymidine incorporation was significantly higher on root tips in the top of tubes, and on root segments and bulk soil in the center of tubes on non-mycorrhizal plants compared to mycorrhizal plants. At the bottom of the tubes, the [³H]-thymidine incorporation was significantly higher on root tips of mycorrhizal plants. Correspondingly, the bacterial biovolumes of rods with dimension 0.28–0.40×1.1–1.6 m, from the bulk soil in the center of tubes and from root segments in the center and top of tubes, and of cocci with a diameter of 0.55–0.78 m in the bulk soil in the center of tubes, were significantly reduced by VAM fungi. The extremely high bacterial biomass (1–7 mg C g^-1 dry weight soil) was significant reduced by mycorrhizal colonization on root segments and in bulk soil. The incorporation of [³H]-thymidine was around one order of magnitude lower compared to other rhizosphere measurements, probably because pseudomonads that did not incorporate [³H]-thymidine dominated the bacterial population. The VAM probably decreased the amount of plant root-derived organic matter available for bacterial growth, and increased bacterial spatial variability by competition. Thus VAM plants seem to be better adapted to compete with the saprophytic soil microflora for common nutrients, e.g., N and P, compared to non-mycorrhizal plants.     

Fungal and bacterial N2O production regulated by soil amendments of simple and complex substrates

Fungal N₂O production results from a respiratory denitrification that reduces NO₃⁻/NO₂⁻ in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N₂O emissions. Effects of substrate quality on fungal and bacterial N₂O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g⁻¹ soil. During periodic measurements of soil N₂O fluxes at 80% soil water-filled pore space and with the supply of KNO₃, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N₂O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N₂O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N₂O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO₂ production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N₂O production, measurements of soil N₂O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N₂O production. As expected, bacterial N₂O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N₂O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N₂O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N₂O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N₂O emissions.     

Do growth yield efficiencies differ between soil microbial communities differing in fungal:bacterial ratios? Reality check and methodological issues

Soil communities dominated by fungi such as those of no-tillage (NT) agroecosystems are often associated with greater soil organic matter (SOM) storage. This has been attributed in part to fungi having a higher growth yield efficiency (GYE) compared to bacteria. That is, for each unit of substrate C utilized, fungi invest a greater proportion into biomass and metabolite production than do bacteria. The assumption of higher fungal efficiency may be unfounded because results from studies in which fungal and bacterial efficiencies have been characterized are equivocal and because few studies have measured microbial GYE directly. In this study, we measured microbial GYE in agricultural soils by following ¹³C-labeled glucose loss, total CO₂-C, and ¹³CO₂-C evolution at 2 h intervals for 20 h in two experiments (differing in N amendment levels) in which the fungal:bacterial biomass ratios (F:B) were manipulated. No differences in efficiency were observed for communities with high versus low F:B in soils with or without added inorganic N. When calculated using ¹³CO₂-C (in contrast to total CO₂-C) evolution, growth yield efficiencies of soils having high and low F:B were 0.69±0.01 and 0.70±0.01, respectively. When soils were amended with N, soils with high and low F:B had growth yield efficiencies of 0.78±0.01 and 0.76±0.01, respectively. Our experiments do not support the widely held assumption that soil fungi have greater growth efficiency than soil bacteria. Thus, claims of greater fungal efficiency may be unsubstantiated and should be evoked cautiously when explaining the mechanisms underlying greater C storage and slower C turnover in fungal-dominated soils.     

Culture-Dependent and Culture-Independent Methods in Evaluation of Emission of Enterobacteriaceae from Sewage to the Air and Surface Water

The number of Enterobacteriaceae, with particular attention given to the presence of Escherichia coli and Klebsiella pneumoniae, was determined in hospital effluents and municipal wastewater after various stages of purification. The emission of these microorganisms to the ambient air near wastewater treatment plant (WWTP) facilities and to the river water, which is a receiver of the WWTP effluent, was also studied using fluorescence in situ hybridization (FISH) and cultivation methods. The number of Enterobacteriaceae determined by cultivation and fluorescence methods in different kinds of sewage sample ranged from 0.5?×?10(3) to 2.9?×?10(6)?CFU/ml and from 2.2?×?10(5) to 1.3?×?10(8) cells/ml, respectively. Their removal rates during treatment processes were close to 99?%, but the number of these bacteria in the WWTP outflow was quite high and ranged from 5.9?×?10(3) to 3.5?×?10(4)?CFU/ml and from 1.1?×?10(5) to 6.1?×?10(5) cells/ml, respectively. In the river water and the air samples, the number of Enterobacteriaceae was also high and ranged from 4.1?×?10(3) to 7.9?×?10(3)?CFU/ml and from 3 to 458?CFU/m(3), respectively. The numbers of these microorganisms obtained from fluorescence and cultivation methods were statistically and significantly correlated; however, the analysis of the studied samples indicated that the FISH method gave values up to 10(3)-fold times greater than those obtained by the cultivation method. From a sanitary point of view, this means that the number of viable fecal bacteria is systematically underestimated by traditional culture-based methods. Thus, the FISH proves to be a method that could be used to estimate bacterial load, particularly in air samples and less contaminated river water.     

Studies of P accumulation in soil/sediment profiles used for large-scale wastewater reclamation

We studied the long‐term accumulation processes and material balances of phosphorus (P) in the soil/sediment profiles of large‐scale effluent recharge basins used for wastewater reclamation by the soil aquifer treatment (SAT) system. The objective was to quantify and clarify the long‐term performance of soil/sediment in the SAT system as a sorbent to filter out P from the recharged effluent. Total P concentration in the soil/sediment profiles of the Shafdan wastewater treatment plant (WWTP) increased over 25 years of operation (1977–2001) by 20–220 mg kg^?1, as a result of adding loads of 0.17–6.2 kg m^?2 of P. Retained P in the 0–2.0 m soil layer increased from 0.06 to 0.31 kg m^?2 with increasing cumulative load of P while the retained percentage gradually decreased from 19 to 5% of the cumulative P load. Accumulation rate of P in the 0–0.15 m horizon in the basins was inversely proportional to recharge time, decreasing from ～28 mg P kg^?1 year^?1 during the first 3 years of operation, to <2.3 mg P kg^?1 year^?1 between the 20th and 25th years of operation. Thus, P content in this horizon approached a steady state after about 10–15 years of effluent recharge under the operational conditions of the Shafdan WWTP. Phosphorus concentration in deeper horizons increased at constant rates of approximately 7.8, 5.9 and 2.9 mg P kg ^?1 year^?1 in the 0.15–0.30, 0.30–0.60 and 1.80 to 2.10‐m horizons, respectively, over the 25 years of effluent recharge. However, the accumulation front of P appears gradually to have moved deeper in the soil profile. In general, this phenomenon may be explained by kinetic limitations to the achievement of full adsorption equilibrium for P between the flowing solution and the solid phase components of the soil. In addition, both the increase of EPC₀(the equilibrium P concentration in solution at which there is no sorption or desorption to or from the soil under the given conditions), caused by long‐term effluent recharge, and gradual decrease of the annual average concentration of P in the effluent input after 1995, may result in the steady‐state level of P in the topsoil of the basin.     

Fecal contamination of Finnish fresh waters in 1962–1984

We investigated fecal contamination of Finnish lakes and rivers on the basis of 132 476 observations of total coliform bacteria (TC), thermotolerant coliform bacteria (ThC) and fecal streptococci (FS) enumerated in a nationwide monitoring programme during the period 1962–1984. The number of analyses of TC was 19 672, of ThC 16 456 and of FS 96 348. The waters were classified into three groups, influenced mainly by municipal wastewaters, industrial wastewaters or diffuse loading. Annual median bacterial concentrations were used for presenting the trends. All the concentrations decreased until about 1976, except TC concentrations in the industrialized and diffuse load areas. Since 1976 the concentrations decreased more slowly or occasionally even increased, especially those of TC. In the whole data the dynamics of bacterial loads (CFU s^?1) were similar to those of annual median bacterial concentrations. The most probable explanations for the declining bacterial concentrations are increase in the human population served by wastewater treatment plants, improved sewage treatment processes, decreased wastewater volumes of the forest industry and decrease in the number of cattle. The monitoring programme continues.     

Modeling soil metabolic processes using isotopologue pairs of position-specific C-labeled glucose and pyruvate

Most organic carbon (C) in soils eventually turns into CO₂ after passing through microbial metabolic pathways, while providing cells with energy and biosynthetic precursors. Therefore, detailed insight into these metabolic processes may help elucidate mechanisms of soil C cycling processes. Here, we describe a modeling approach to quantify the C flux through metabolic pathways by adding 1-¹³C and 2,3-¹³C pyruvate and 1-¹³C and U-¹³C glucose as metabolic tracers to intact soil microbial communities. The model calculates, assuming steady-state conditions and glucose as the only substrate, the reaction rates through glycolysis, Krebs cycle, pentose phosphate pathway, anaplerotic activity through pyruvate carboxylase, and various biosynthesis reactions. The model assumes a known and constant microbial proportional precursor demand, estimated from literature data. The model is parameterized with experimentally determined ratios of ¹³CO₂ production from pyruvate and glucose isotopologue pairs. Model sensitivity analysis shows that metabolic flux patterns are especially responsive to changes in experimentally determined ¹³CO₂ ratios from pyruvate and glucose. Calculated fluxes are far less sensitive to assumptions concerning microbial chemical and community composition. The calculated metabolic flux pattern for a young volcanic soil indicates significant pentose phosphate pathway activity in excess of pentose precursor demand and significant anaplerotic activity. These C flux patterns can be used to calculate C use efficiency, energy production and consumption for growth and maintenance purposes, substrate consumption, nitrogen demand, oxygen consumption, and microbial C isotope composition. The metabolic labeling and modeling methods may improve our ability to study the biochemistry and ecophysiology of intact and undisturbed soil microbial communities.     

Measurement of triclosan in water using a magnetic particle enzyme immunoassay

A sensitive magnetic particle-based immunoassay to determine triclosan [5-chloro-2-(2,4-dichlorophenoxy)phenol] in drinking water and wastewater was developed. Rabbit antiserum was produced by immunizing the rabbit with 6-[5-chloro-2-(2,4-dichlorophenoxy)phenoxy]hexanoic acid-keyhole limpet hemocyanin. Horseradish peroxidase was conjugated with 4-[3-bromo-4-(2,4-dibromophenoxy)phenoxy]butyric acid via N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). The triclosan antibody was coupled to magnetic particles via the NHS/EDC reaction. The antibodies were able to recognize some structurally related polybrominated biphenyl ethers but did not recognize various common pollutants that were less similar to the hapten. The ELISA could detect triclosan in standard solution (25% methanol/H2O v/v) at 20 ppt and its metabolite, methyl-triclosan, at 15 ppt. Water samples from different treatment stages were prepared to contain 25% methanol and analyzed directly without any sample extraction or preconcentration. The results showed that recoveries were >80% and the % CV was <10%, demonstrating the assay was both accurate and precise. Application of the triclosan ELISA to water treatment plants showed that tap water at various purification stages had low concentrations of triclosan (<20 ppt) and required an increased sample size for appropriate detection and measurement. Application of ELISA to the wastewater treatment plants (WWTP) demonstrated high concentrations of triclosan (in general, >3000 ppt in water entering the WWTP) with the levels decreasing as the water proceeded through the processing plant (<500 ppt at outflow sewage). The ELISA measurement was shown to be equivalent to the more specific GC-MS analysis on a number of wastewater treatment samples with a high degree of correlation, with the exception of a few samples with very high triclosan concentrations (>5000 ppt). Measurement of methyl-triclosan (in WWTP) using GC-MS demonstrated the levels of this compound to be low. In summary, a rapid, sensitive, accurate, and precise magnetic particle-based immunoassay has been developed for triclosan analysis, which can serve as a cost-effective monitoring tool for various water samples.     

Fungal/bacterial ratios in grasslands with contrasting nitrogen management

It is frequently hypothesised that high soil fungal/bacterial ratios are indicative for more sustainable agricultural systems. Increased F/B ratios have been reported in extensively managed grasslands. To determine the shifts in fungal/bacterial biomass ratio as influenced by grassland management and to find relations with nitrogen leaching potential, we sampled a two-year-old field experiment at an organic experimental farm in the eastern part of The Netherlands. The effect of crop (grass and grass-clover), N application rate (0, 40, 80, ) and manure type (no manure, farm yard manure and slurry) on the F/B ratio within three growing seasons was tested, as well as relations with soil and crop characteristics, nitrate leaching and partial N balance. Biomass of fungi and bacteria was calculated after direct counts using epifluorescence microscopy. Fungal and bacterial biomass and the F/B ratio were higher in grass than in grass-clover. The F/B ratio decreased with increasing N application rate and multiple regression analysis revealed a negative relationship with pH. Bacterial activity (measured as incorporation of [³H]thymidine and [¹⁴C]leucine into bacterial DNA and proteins) showed the exact opposite: an increase with N application rate and pH. Leaching increased with N application rate and was higher in grass-clover than in grass. Partial N balance was more positive at a higher N application rate and showed an inverse relationship with fungal biomass and F/B ratio. We conclude that the fungal/bacterial biomass ratio quickly responded to changes in management. Grasslands with higher N input showed lower F/B ratios. Grass-clover had a smaller fungal biomass and higher N leaching than grass. In general, a higher fungal biomass indicated a lower nitrogen leaching and a more negative partial N balance (or smaller N surplus), but more observations are needed to confirm the relationship between F/B ratio and sustainability.     

极浅型潜流人工湿地用于污水处理和风沙土改良的中试研究

[目的]就利用极浅型潜流人工湿地同步处理污水和改良风沙土的可行性进行中试,为后续相关工程的设计、建设和运行提供科学依据。[方法]以污水处理厂初沉池出水为原水,以取自毛乌素沙漠的风沙土为湿地填料,以高羊茅、黑麦草和狗牙根为湿地栽培植物,研究并分析湿地对水中污染物的去除效果和在风沙土的富集特征。[结果]经过15个月的运行,各湿地对污水中COD,TN以及TP的平均去除率分别高于原土68.75%,70.09%和62.74%。运行完毕后,各湿地风沙土较原土有机质、全氮含量显著增加(p0.05),全磷含量虽呈增加趋势但增幅不显著(p0.05),其中风沙土有机质、全氮及全磷含量分别较原土增加了10.64~18.77,8.74~13.98,0.34~0.55倍。[结论]利用极浅型潜流人工湿地同步处理污水和改良风沙土可行。它既能高效净化污水,又可快速提升风沙土的生产力。     

A new method for determining the metabolic activity of specific bacterial populations in soil using tritiated leucine and immunomagnetic separation

 A new assay, using immunomagnetic separation and uptake of tritiated leucine ([³H]-Leu), was developed for measuring the in situ metabolic activity of specific bacterial populations in soil. Such assays are needed to assess the role individual species play in diverse microbial soil communities. The method was optimized using Pseudomonas putida KT2440 : :Tc⁺/TOL::gfp inoculated into soil microcosms. Inoculated soil samples were incubated with [³H]-Leu followed by an immunomagnetic separation to recover the target bacteria. Radiolabel incorporated by the target bacteria was then measured. Incubation time with immunomagnetic beads was not critical for optimal target cell recovery, but samples needed to be washed at least 5 times during the immunomagnetic separation to reduce unspecific binding of the indigenous soil bacteria to the magnetic beads. Soil absorption of the polyclonal antibody further reduced this unspecific binding, resulting in <1% contamination by indigenous soil bacteria relative to numbers of recovered target cells. The assay was tested by investigating the effect of different incubation temperatures on the metabolic activity of the target cells. As expected, a linear relationship between activity and temperature was observed, demonstrating the sensitivity of the assay. The method was applied to compare activities of the target strain in bulk soil and in the rhizosphere of barley. Contrary to what was anticipated, no significant difference in metabolic activity was observed. Received: 4 November 1999     

Ciprofloxacin Resistance in Domestic Wastewater Treatment Plants

The potential of domestic wastewater treatment plants to contribute for the dissemination of ciprofloxacin-resistant bacteria was assessed. Differences on bacterial counts and percentage of resistance in the raw wastewater could not be explained on basis of the size of the plant or demographic characteristics of population served. In contrast, the treated effluent of the larger plants had significantly more heterotrophs and enterobacteria, including ciprofloxacin-resistant organisms, than the smaller (p?<?0.01). Moreover, longer hydraulic retention times were associated with significantly higher percentages of resistant enterobacteria in the treated effluent (p?<?0.05). Independently of the size or type of treatment used, domestic wastewater treatment plants discharged per day at least 10¹⁰–10¹⁴ colony forming units of ciprofloxacin-resistant bacteria into the receiving environment.     

Sorption of Polymeric Quaternary Ammonium Compounds to Humic Acid

Polymeric quaternary ammonium salts or polyquaterniums are used not only in the water and wastewater industry but also in cosmetics. The former have been extensively studied with sorption to wastewater treatment plant (WWTP) biosolids an important factor in their fate, mitigating release to the environment. Compounds of cosmetic origin have not received the same scrutiny as those used in other applications despite differences in their structure, characteristics and properties such as toxicity. In this work, the sorption of selected polyquaterniums found in cosmetics to treated humic acid, employed as a surrogate for WWTP biosolids, is investigated. For comparison, the sorption of poly(diallyldimethylammonium chloride) (poly(DADMAC)), a polyquaternium commonly used as a flocculant, and cetyl pyridinium chloride, a monomeric quaternary ammonium compound, are also determined. The humic acid/water partition coefficients (K _D) of cetyl pyridinium chloride (52,000?L?kg^?1) and poly(DADMAC) (2,200?L?kg^?1) were greater than those of the polyquaterniums found in cosmetics (200 to 1,200?L?kg^?1). Assumptions of partitioning mainly to biosolids and substantial removal relative to the influent concentration of WWTPs may therefore not be valid.     

Short-term effects in soil microbial community following agronomic application of olive mill wastewaters in a field of olive trees

Olive mill wastewater (OMW) creates a disposal problem. The large amounts generated, combined with the high phenol and chemical oxygen demand concentrations, are the main difficulties in finding a solution for the management of these wastewaters. We investigated the short-term effect of spreading OMW on the soil surface of an olive grove on the soil microbial communities. Analyse of ester-linked fatty acid methyl ester (EL-FAME) were used to assess variation in soil microbial community structure after agronomic application of OMW. EL-FAME analysis showed significant shifts of specific groups of fatty acids 30 days after application of OMW to a field of olive trees at rates of 0 (control soil), 30, 60, 100, and 150 m³ ha⁻¹ of OMW. In particular, the branched saturated fatty acids indicative of Gram-positive bacteria decreased and the unsaturated fatty acids commonly found in Gram-negative bacteria and fungi increased. The fungal/bacterial ratio measured increased significantly with increasing OMW. Lower cy19/18:1ω7c and cy17/16:1ω7c ratios were found in the amended soil than the control soil, and we interpret that as an indication that nutrient availability may be more limiting in the control soil. Similarly, the relative abundances of monounsaturated fatty acids increased with added OMW, and this is consistent with the presence of high substrate availability in OMW-treated soil. Principal Components Analysis of the FAME profiles showed discrimination between the control soil and OMW amended soil. Differences in fatty acid profiles between OMW-treated soil and control soil suggests that amendment of soil with OMW favors specific groups of organisms. To our knowledge, this is the first report of alterations in the FAME profile in soils due to agronomic application of OMW.     

Shifts in microbial community and water-extractable organic matter composition with biochar amendment in a temperate forest soil

Biochar amendment in soil has been proposed as a carbon sequestration strategy which may also enhance soil physical and chemical properties such as nutrient and water holding capacity as well as soil fertility and plant productivity. However, biochar may also stimulate microbial activity which may lead to increased soil CO₂ respiration and accelerated soil organic matter (OM) degradation which could partially negate these intended benefits. To investigate short-term soil microbial responses to biochar addition, we conducted a 24 week laboratory incubation study. Biochar produced from the pyrolysis of sugar maple wood at 500 °C was amended at concentrations of 5, 10 and 20 t/ha in a phosphorus-limited forest soil which is under investigation as a site for biochar amendment. The cumulative soil CO₂ respired was higher for biochar-amended samples relative to controls. At 10 and 20 t/ha biochar application rates, the concentration of phospholipid fatty acids (PLFAs) specific to Gram-positive and Gram-negative bacteria as well as actinomycetes were lower than controls for the first 16 weeks, then increased between weeks 16–24, suggesting a gradual microbial adaptation to altered soil conditions. Increases in the ratio of bacteria/fungi and lower ratios of Gram-negative/Gram-positive bacteria suggest a microbial community shift in favour of Gram-positive bacteria. In addition, decreasing ratios of cy17:0/16:1ω7 PLFAs, a proxy used to examine bacterial substrate limitation, suggest that bacteria adapted to the new conditions in biochar-amended soil over time. Concentrations of water-extractable organic matter (WEOM) increased in all samples after 24 weeks and were higher than controls for two of the biochar application rates. Solution-state ¹H NMR analysis of WEOM revealed an increase in microbial-derived short-chain carboxylic acids, lower concentrations of labile carbohydrate and peptide components of soil OM and potential accumulation of more recalcitrant polymethylene carbon during the incubation. Our results collectively suggest that biochar amendment increases the activity of specific microorganisms in soil, leading to increased CO₂ fluxes and degradation of labile soil OM constituents.     

Bacterial and fungal contributions to soil nitrogen cycling under Douglas fir and red alder at two sites in Oregon

A study was conducted at two experimental tree plantations in the Pacific Northwest to assess the roles of bacteria and fungi in nitrogen (N) cycling. Soils from red alder (Alnus rubra) and Douglas-fir (Pseudotsuga menziesii) plots in low- (H.J. Andrews) and high- (Cascade Head) productivity stands were sampled in 2005 and 2006. Fungal:bacterial ratios were determined using phospholipid fatty acid (PLFA) profiles and quantitative (Q)-PCR. Ratios from these two molecular methods were highly correlated and showed that microbial biomass varied significantly between the two experimental sites and to a lesser extent between tree types with fungal:bacterial biomass ratios lower in more N-rich plots. ¹⁵N isotope dilution experiments, with ammonium (NH₄⁺) and nitrate (NO₃^?), were paired with antibiotics that blocked bacterial (bronopol) and fungal (cycloheximide) protein synthesis. This modified isotope dilution technique was used to determine the relative contribution of bacteria and fungi to net N mineralization and gross rates of ammonification and nitrification. When bacterial protein synthesis was blocked NH₄⁺ consumption and nitrification rates decreased in all treatments except for NH₄⁺ consumption in the Douglas-fir plots at H.J. Andrews, suggesting that prokaryotic nitrifiers are a major sink for mineral NH₄⁺ in forest soils with higher N availability. Cycloheximide consistently increased NH₄⁺ consumption, however the trend was not statistically significant. Both antibiotics additions also significantly increased gross ammonification, which may have been due to continued activity of extra- and intracellular enzymes involved in producing NH₄⁺ combined with the inhibition of NH₄⁺ assimilation into proteins. The implication of this result is that microorganisms are likely a major sink for soil dissolved organic N (DON) in soils.     

Recovery of aquatic bacterial populations in a stream after cessation of chemical pollution

Tims Branch was studied for 1 yr to determine the effects of an annual loading of about 229 900 kg of various chemicals on the bacterial populations of a small fast flowing stream. The 10 yr discharge of chemical waste to this stream was terminated in 1973 and a 2 yr study of the stream's recovery was begun. Upper Three Runs, an adjacent nonpolluted stream, was used as a control. Total culturable bacterial counts, per cent chromagenic bacteria and diversity were used as indices of recovery. Plate count agar was used for culturing aquatic bacteria present in both systems. Diversity in both systems was more affected by seasonal temperatures than the presence of chemical pollution. Mean total counts dropped from 187 × 10³ in the polluted stream to approximate those of the control stream (170 × 10³) following cessation of pollution. In the chemically affected stream, per cent chromagens increased from 15% during pollution to 25% approximately 1 yr after pollution was stopped. The latter percentage was comparable to that of the control stream (28%) during the period of study.     

Root-associated bacteria containing 1-aminocyclopropane-1-carboxylate deaminase improve growth and nutrient uptake by pea genotypes cultivated in cadmium supplemented soil

The effect of inoculation with Pseudomonas brassicacearum Am3, Pseudomonas marginalis Dp1 and Rhodococcus sp. Fp2 containing 1-aminocyclopropane-1-carboxylate deaminase (ACCD) on growth and uptake of N, P, K, Ca, S, Fe and Cd in shoots of pea (Pisum sativum) genotypes VIR188, VIR1658, VIR3429 and VIR4488 was studied in pot experiment with non-polluted and Cd-supplemented (10 mg Cd kg⁻¹) sod-podzolic soil. The growth-promoting effect of bacteria depended on plant genotype and bacterial strain. Only Rhodococcus sp. Fp2 had no ACCD activity in vitro in the presence of Cd and did not stimulate pea growth in Cd-supplemented soil. Inoculation with bacteria counteracted the Cd-induced inhibition of nutrient uptake by plants probably through stimulation of root growth and enhancement of nutrient uptake processes. Nutritional effects of the bacteria were specific with respect to the nutrient.