Anaerobic sediment potential acidification and metal release risk assessment by chemical characterization and batch resuspension experiments

Background, Aim and Scope  Sediments act as a sink for toxic substances (heavy metals, organic pollutants) and, consequently, dredged materials often contain pollutants which are above safe limits. In polluted anaerobic sediments, the presence of sulphides and redox potential changes creates a favorable condition for sulphide oxidation to sulphate, resulting in potential toxic metal release. The oxidation reaction is catalyzed by several microorganisms. Some clean up measures, such as dredging, can initiate the process. The aim of the present work is to assess the acidification and metal release risk in the event of sediment dredging and also to compare two different acid base account techniques with the resuspension results. The oxidation mechanism by means of inoculation with an Acidithiobacillus ferrooxidans strain was also evaluated. Methodology  The sediments were chemically characterized (pH; organic oxidizable carbon; acid volatile sulphides; total sulphur; moisture; Cr, Cu and Zn aqua regia contents). A metal sequential extraction procedure (Community Bureau of Reference, BCR technique) was applied to calculate the Acid Producing Potential (APP) and Acid Consuming Capacity (ACC) of the sediment samples through Fe, Ca²⁺ and SO₄ ²⁻ measurements. The acid base account was also performed by the Sobek methodology (Acid producing potential — AP — calculated with total sulphur and neutralization potential — NP — by titration of the remaining acid after a reaction period with the sample). Fresh sediments were placed in agitated shake flasks and samples were taken at different times to evaluate pH, SO₄ ²⁻ and Cr, Cu, Zn and Fe²⁺ concentration. Some of the systems were inoculated with an Acidithiobacillus ferrooxidans strain to assess the biological catalysis on sulphide oxidation. Results  Sediment chemical characterization showed high organic matter content (5.4–10.6%), total sulphur (0.36–0.86%) and equivalent CaCO₃ percentages (4.5–8%). pH was neutral-alkaline for all of the samples. AVS content was high except for sample 5. The acid base account obtained with the two methods gave different results for the acid generating risk of the samples. A decrease of 0.4 to 3.1 pH units was measured in the agitated shake flasks. In all of the systems, sulphate concentration increased (2,100–2,200 mg L⁻¹ to 2,500–3,000 mg L⁻¹), and positively correlated with the initial total sulphur content of the samples in the inoculated flasks. Cu and Cr in solution were not detected in most of the sampling occasions (<0.5 mg Cu L⁻¹ and <0.5 mg Cr L⁻¹). Zn reached high concentrations (up to 11.8 mg L⁻¹). For every system — except sediment 1 — the lowest pH registered was similar in comparison to inoculated and control systems. The inoculation effect was mostly evidenced in the systems by a higher sulphate release rate compared to the control systems. Discussion  The BCR method categorized all of the samples as potentially acid generating material. The Sobek method using NPR (NP/AP) criteria classified sample 3 as a possible acid generator and samples 1, 2 and 5 with a low acid generation potential. Despite this, all the samples acidified the media in the kinetic tests in at least one of the conditions employed in this work. It would seem that NPR and NNP (NP-AP) risk classification criteria should not be directly used with anaerobic sediments. Appropriate classification levels for sediments should be developed considering the different sulphide reactivity between rock and sediments. Sediment oxidation can cause acidification, which is partially explained by sulphide oxidation. In the samples studied, we found a positive correlation between sulphate increase in solution after oxidation and total sulphur content in the inoculated systems. Significant amounts of Zn could be released to solution while Cr and Cu remained insoluble despite the pH decrease observed. The low Cu and Cr mobility could be explained by the very low solubility of their hydroxides and high affinity for organic matter and iron oxides/hydroxides that might form during sediment oxidation. Dredged sediment management and disposal should be carefully planned. Conclusions  All of the sediment samples lowered the pH media in the laboratory batch resuspension experiments. However, both risk classification criteria (NNP, NPR) from Sobek acid base account were not able to predict the samples’ behavior as accurately as the BCR derived base account. The inoculation effect was mostly associated with a higher sulphate release and not to a lower pH due to acid base equilibrium. Recommendations and Perspectives  Appropriate risk classification levels for sediments should be developed considering the different sulphide reactivity between rock and sediments. ESS-Submission Editor: Dr. Sabine Ulrike Gerbersdorf (sug@st-andrews.ac.uk)     

A new sediment contact assay to assess particle-bound pollutants using zebrafish (<Emphasis Type="Italic">danio rerio</Emphasis>) embryos

Goal, Scope and Background. Based on a bioassay battery covering only primary producers and consumers as well as degraders, the potential ecological hazard of sediments to vertebrates cannot be estimated comprehensively. Therefore, there is an urgent need to develop and standardize integrated vertebrate-based test systems for sediment investigation strategies. Whereas vertebratebased in vitro systems have frequently been used for the investigation of aqueous samples, there is a significant lack of whole sediment assays. Thus, the purpose of the present study was: (1) to develop a rapid and reliable, but comprehensive method to investigate native sediments and particulate matters without preceding extraction procedures; (2) to compare the hazard potential of solid phase sediments to the effects of corresponding pore waters and organic extracts in order to characterize the bioavailability of the particle-bound pollutants; and (3) to relatively evaluate the embryotoxic effects of sediments from the catchment areas of the rivers Rhine, Neckar and Danube. Methods (or Main Features).  To investigate the toxicity of sediment samples on vertebrates, the standard embryo toxicity test with the zebrafish (Danio rerio; Hamilton-Buchanan 1922) according to DIN 38415-6 was modified with respect to exposure scheme and toxicological endpoints. Sediments from the catchment area of the Neckar River were assessed using pore waters, acetonic extracts and native sediments in order to get inside into the potential bioavailability of particle-bound pollutants. A comprehensive test protocol for the investigation of native sediments in the embryo toxicity test with the zebrafish is presented. Results and Discussion.  The fish embryo assay with Danio rerio can be carried out with both aqueous and organic sediment extracts as well as native (whole, solid phase) sediment samples. Elongation of exposure time from 48 to up to 196 h significantly increased the mortality. Using the fish egg assay with native sediments, a broad range of embryotoxic effects could be elucidated, including clear-cut dose-response curves for the embryotoxic effects of contaminated sediments; in contrast, absence of embryotoxic effects could be demonstrated even for the highest test concentrations of unpolluted sediments. With native sediments, embryotoxicity was clearly higher than with corresponding pore waters, thus corroborating the view that — at least for fish eggs — the bioavailability of particle-bound lipophilic substances in native sediments is higher than generally assumed. The relative ranking of sediment toxicity was identical using both native sediments and sediment extracts, EC₂₀ values of the latter, however, being eight time lower higher than with the native sediments. A comparison of the embryo toxic effects of samples from the Neckar area with locations along the Rhine and Danube rivers elucidated a broad range of results, thus indicating different levels of contamination. Conclusions.  A modified protocol of the zebrafish embryo test allows the assessment of sediment toxicity in both aqueous extracts and native sediments. The isolated investigation of pore waters may result in a clear-cut underestimation of the bioavailability of lipophilic particle-bound substances (as determined by native sediments). Recommendations and Perspectives.  The zebrafish embryo test with native (whole, solid phase) sediments appears very promising for the evaluation of the bioavailable fraction of lipophilic particle-bound substances and can therefore be recommended for the evaluation of vertebrate toxicity in tiered sediment test strategies and dredging directives such as the HABAB-WSV. Whereas acetone extracts may be tested as a rough estimation of embryotoxicity, native sediment samples will provide a more comprehensive and realistic insight into the bioavailable hazard potential     

Impact of heavy metal amended sewage sludge on forest soils as assessed by bacterial and fungal biosensors

Bacterial and fungal bioluminescence-based biosensors were used as indicators of potential heavy metal toxicity to microorganisms in the needle litter of a mature Pinus radiata forest under heavy metal contaminated sewage sludge. Sewage sludge was amended with increasing concentrations of Cu, Ni and Zn and applied to the surface of a mature P. radiata forest. The response of the bacterial and fungal biosensors to soluble Cu, Ni and Zn in needle litter extracts was investigated. The bioluminescence response of the bacterial biosensor Escherichia coli HB101 pUCD607 declined as water-soluble Zn concentrations increased. The effective concentrations that gave a 50% reduction in bioluminescence (EC₅₀ values) for water-soluble Zn and total litter Zn were 1.3 mg l⁻¹ and 3700 mg kg⁻¹, respectively. The bioluminescence response of the fungal biosensor Armillaria mellea declined as soluble Cu concentrations increased. The EC₅₀ values for water-soluble Cu and total litter Cu were 0.12 mg l⁻¹ and 540 mg kg⁻¹, respectively. No decline in bioluminescence was noted for either the bacterial or fungal biosensor on exposure to increasing concentrations of water-soluble Ni. The use of a combination of bacterial and fungal biosensors offers a rapid and sensitive tool for assessing toxicity of heavy metals to microorganisms and, thus, elucidating the environmental impact of contaminants in sewage sludge on litter dwelling microorganisms.     

微生物制剂对肉鸡屠宰废水生物降解效果的研究

采用沼泽红假单胞菌、保加利亚乳杆菌、酵母菌和有效微生物群组成不同组合，以pH值、浑浊度、氨氮浓度、亚硝酸盐氮含量、BOD₅值和COD_mn值为考察指标，研究不同微生物制剂对肉鸡屠宰废水生物降效果，结果表明：沼泽红假单胞菌、保加利亚乳杆菌和有效微生物群以25∶25∶1的比例组成的复合微生物制剂，以0.01%为添加量时效果最好。     

Effects of long-term mineral fertilization on microbial biomass,microbial activity,and the presence of <Emphasis Type="Italic">r</Emphasis>- and <Emphasis Type="Italic">K</Emphasis>-strategists in soil

Long-term effects of mineral fertilization on microbial biomass C (MBC), basal respiration (R _B), substrate-induced respiration (R _S), β-glucosidase activity, and the r–K-growth strategy of soil microflora were investigated using a field trial on grassland established in 1969. The experimental plots were fertilized at three rates of mineral N (0, 80, and 160 kg ha⁻¹ year⁻¹) with 32 kg P ha⁻¹ year⁻¹ and 100 kg K ha⁻¹ year⁻¹. No fertilizer was applied on the control plots (C). The application of a mineral fertilizer led to lower values of the MBC and R _B, probably as a result of fast mineralization of available substrate after an input of the mineral fertilizer. The application of mineral N decreased the content of C extracted by 0.5 M K₂SO₄ (C _ex). A positive correlation was found between pH and the proportion of active microflora (R _S/MBC). The specific growth rate (μ) of soil heterotrophs was higher in the fertilized than in unfertilized soils, suggesting the stimulation of r-strategists, probably as the result of the presence of available P and rhizodepositions. The cessation of fertilization with 320 kg N ha⁻¹ year⁻¹ (NF) in 1989 also stimulated r-strategists compared to C soil, probably as the result of the higher content of available P in the NF soil than in the C soil.     

碱蓬浮床对海水养殖尾水的修复效果

[目的]利用碱蓬浮床对富营养化、高盐碱的海水养殖尾水进行修复,为治理严重的海洋环境问题提供新的方法。[方法]通过设计一种兼具物理、化学、生物协同作用的新型组合式碱蓬(Suaeda salsa)浮床,研究其对富营养化海水养殖尾水中总氮(TN)、总磷(TP)、高锰酸钾指数(COD_(Mn))、氨氮(NH_4~+-N)的去除效果,并与传统浮床进行对比。[结果]组合式浮床对TN,NH_4~+-N,TP,COD_(Mn)的去除率分别为62.14%,93.94%,73.05%和60.91%,较传统浮床去除率分别提高了27.36%,21.27%,19.51%,12.43%。[结论]组合式浮床对富营养化、高盐碱的海水养殖尾水具有明显的修复能力。     

Emissions of nitrous oxide from a constructed wetland using a groundfilter and macrophytes in waste-water purification of a dairy farm

 In less populated rural areas constructed wetlands with a groundfilter made out of the local soil mixed with peat and planted with common reed (Phragmites australis) are increasingly used to purify waste water. Particularly in the rhizosphere of the reed, nitrification and denitrification processes take place varying locally and temporally, and the question arises to what extent this type of waste-water treatment plant may contribute to the release of N₂O. In situ N₂O measurements were carried out in the two reed beds of the Friedelhausen dairy farm, Hesse, Germany, irrigated with the waste water from a cheese dairy and 70 local inhabitants (12 m³ waste water or 6 kg BOD₅ or 11 kg chemical O₂ demand (COD_Mn) day^–1). During November 1995 to March 1996, the release of N₂O was measured weekly at 1 m distances using eight open chambers and molecular-sieve traps to collect and absorb the emitted N₂O. Simultanously, the N₂O trapped in the soil, the soil temperature, as well as the concentrations of NH₄ ⁺-N, NO₃ ^–-N, NO₂ ^–-N, water-soluble C and the pH were determined at depths of 0–20, 20–40 and 40–60 cm. In the waste water from the in- and outflow the concentrations of COD_Mn, BOD₅, NH₄ ⁺-N, NO₃ ^–-N, NO₂ ^–-N, as well as the pH, were determined weekly. Highly varying amounts of N₂O were emitted at all measuring dates during the winter. Even at soil temperatures of –1.5  °C in 10 cm depth of soil or 2  °C at a depth of 50 cm, N₂O was released. The highest organic matter and N transformation rates were recorded in the upper 20 cm of soil and in the region closest to the outflow of the constructed wetland. Not until a freezing period of several weeks did the N₂O emissions drop drastically. During the period of decreasing temperatures less NO₃ ^–-N was formed in the soil, but the NH₄ ⁺-N concentrations increased. On average the constructed wetlands of Friedelhausen emitted about 15 mg N₂O-N inhabitant equivalent^–1 day^–1 during the winter period. Nitrification-denitrification processes rather than heterotrophic denitrification are assumed to be responsible for the N₂O production. Received: 28 October 1998     

Phenylpyrazoles impact on Folsomia candida (Collembola)

Effects of the broad-spectrum insecticide fipronil were investigated on a non-target insect living in the soil, the springtail Folsomia candida Willem. Fipronil induced a significant reduction in juvenile production (PNEC = 250 μg kg⁻¹ dry soil), which seemed to be linked with an impact on the first stages of springtail development: juveniles and 7-day-old adults. These young organisms have a thinner integument, a smaller mass body and a weaker detoxification efficiency and were more sensitive than adults (14 days old) to fipronil and phenylpyrazole derivatives. Contact toxicity for juveniles was measured (LC₅₀(96 h)) giving the following values: fipronil, 450 μg l⁻¹; sulfone-fipronil, 430 μg l⁻¹; sulfide-fipronil, 160 μg l⁻¹. F. candida organisms were able to avoid contaminated food because phenylpyrazoles decreased food appetency. However, F. candida could bioaccumulate fipronil through trans-tegumental penetration (BAF_96 h = 160) and its high biotransformation rate inside springtail bodies (1 ng fipronil metabolized day⁻¹ individual⁻¹) was suspected to increase this process. Under natural conditions, phenylpyrazoles risk assessment on springtails seems to be weak due to their capacity of avoiding high contaminated zones and their biochemical tolerance to this class of insecticides.     

Application of a new sediment contact test with <Emphasis Type="Italic">Myriophyllum aquaticum</Emphasis> and of the aquatic <Emphasis Type="Italic">Lemna</Emphasis> test to assess the sediment quality of Lake Skadar

Goal, Scope and Background  Situated in the transboundary belt between Montenegro and Albania, Lake Skadar is the largest freshwater reservoir in Southeastern Europe. Because of the wide range of endemic, rare or endangered plant and animal species it supports, Lake Skadar and its extensive adjacent wetlands are internationally recognised as a site of significance and importance (Ramsar site). Within the last 10 to 20 years, Lake Skadar was exposed to intensive pollution. For the assessment of the ecotoxic load of the sediments sampled in Lake Skadar, a triad approach was recently applied. Overall, a complex spectrum of ecotoxic loads was elucidated. The aim of the present study was to use plant-based bioassays for assessing the sediment quality of Lake Skadar in order to facilitate and complement the triad test battery. The newly developed sediment contact test with Myriophyllum aquaticum and the aquatic growth inhibition test with Lemna minor were applied to native sediments and pore water, respectively, allowing the investigation of different toxicity-effects caused by particle-bound pollutants as well as pollutants in the interstitial water. This investigation is the first application of the novel sediment contact test with Myriophyllum aquaticum to lake sediments. Methods  Sediment samples were taken from nine selected sites at Lake Skadar and investigated by the sediment contact assay with Myriophyllum aquaticum. The pore water was extracted from these sediment samples to be analysed in the aquatic growth inhibition test with Lemna minor. The results of the sediment contact tests were compared with each other and with those of the aquatic growth inhibition test. Results and Discussion  Both applied macrophyte biotests revealed distinct changes in the growth behaviour of the two macrophytes subsequent to the exposure to the investigated natural sediments of Lake Skadar. The Myriophyllum sediment contact test revealed significant toxicity in the sediment samples from Radus and Kamenik, whereas the aquatic Lemna test showed inhibition effects for the samples from Sterbeq, Plavnica and Kamice. Data obtained with the newly developed Danio rerio contact test and the Arthrobacter globiformis contact test confirmed the Myriophyllum results. Analyses of the heavy metal content in the sediments revealed low or moderate contamination levels. Correlation analyses between the content of heavy metals in the sediments and growth inhibition of Myriophyllum aquaticum showed a significant correlation between Cr concentrations and growth inhibition. Comparable findings are available for a German river system. In contrast, no significant correlation between inhibition rates and concentration of metals could be observed with Lemna minor. Conclusions  It was shown that the newly developed sediment contact test with Myriophyllum aquaticum is applicable to lake sediments. In both the sediment contact test with Myriophyllum aquaticum on whole sediments and the aquatic growth inhibition test with Lemna minor on pore water, plant growth was influenced by the natural sediments and its components. Therefore, both test systems were found to be suitable for the detection of phytotoxic effects upon exposure to sediments. Myriophyllum aquaticum as test organism of the contact test grows directly in the sediment without an additional water-layer. Thus, it is able to detect toxicity caused by particle-bound phytotoxic substances as well as pore water-related contamination, while the floating Lemna minor can only detect effects emanating from pore water. Significant differences of the results were observed between these two test systems and, accordingly, the two different exposure scenarios. Hence, none of the tests can replace the other one and, as a consequence, both should be included into a test battery for the assessment of sediment toxicity. Recommendations and Perspectives  Both plant assays were shown to be reliable tools for the evaluation of the eco-toxicological risk potentials of pore water and solid-phase sediment. They should become a complement to the standardised test battery generally used for comprehensive hazard assessment. ESS-Submission Editor: Dr. Ulrike Kammann (ulrike.kammann@ifo.bfa-fisch.de)     

Sediment,Perch (Perca fluviatilis L.) and Bottom Fauna as Indicators of Effluent Discharged from the Pulp and Paper Mill Complex at Kemi,Northern Finland

This paper compares the long-term data of EOX (Extractable Organic Halogens) determinations in sediments, trichloroguaiacol, i.e. 3,4,5-trichloroguaiacol and 4,5,6-trichloroguaiacol, determinations in the bile of perch (Perca fluviatilis L.) and changes in bottom fauna communities (Pontoporeia affinis L., Mesidotea entomon L., Valvata spp. L.) during 1994–2003. The study area is located at Kemi, on the Gulf of Bothnia, Northern Finland, and is affected by effluent discharged from the pulp and paper mills of Stora Enso Oyj and Oy Metsä-Botnia Ab. The Kemi municipal sewage plant, and River Kemijoki also impose a load in the same area. The main purpose of the study was to assess whether the state of the exposed water ecosystem has improved on the basis of the concentrations of organochlorine compounds in sediment and in perch, due to the modernisation of the wastewater treatment at the mills. In addition, we also investigated whether the changes in the bottom fauna community indicate an improvement in the purification of the effluents. The results indicate that the concentrations of trichloroguaiacols (–1) in perch reflect the decreased discharges of chlorinated substances from the mills. The EOX concentrations in bottom sediments have also decreased to below 50 of Cl g^–1 in most parts of the study area, which is in good agreement with the reduction in the discharge of organochlorines. According to the long-term changes in bottom fauna, the reduction in the discharge of organochlorine compounds due to modernisation of the pulp mills is reflected in a recovery of the benthic fauna, especially Valvata spp. L. and Pontoporeia affinis L.     

Chronic toxicity of unresolved complex mixtures (UCM) of hydrocarbons in marine sediments

Background, Aim and Scope  Unresolved complex mixtures (UCM) of hydrocarbons, containing many thousands of compounds which cannot be resolved by conventional gas chromatography (GC), are common contaminants of sediments but little is known of their potential to affect sediment-dwelling organisms. Evidence exists for reduced health status in mussels, arising from aqueous exposure to aromatic UCM components acting through a narcotic mode of action. However, UCM contaminants in sediments may not be sufficiently bioavailable to elicit toxic effects. The aim of our study was therefore to measure the sublethal effects of chronic exposure to model UCM-dominated oils at environmentally realistic concentrations and compare this to effects produced by a UCM containing weathered crude oil. A further aim was to determine which, if any, fractions of the oils were responsible for any observed toxicity. Materials and Methods  Whole oils were spiked into estuarine sediment to give nominal concentrations of 500 μg g⁻¹ dry weight. Juveniles of the estuarine amphipod Corophium volutator were exposed to the contaminated sediment for 35 days and their survival, growth rate and reproductive success quantified. Using an effect-directed fractionation approach, the oils were fractionated into aliphatic and two aromatic fractions by open column chromatography and their toxicity assessed by further chronic exposures using juvenile C. volutator. Results  The growth rates of amphipods were reduced following exposure to the oils although this was only statistically significant for the weathered oil; reproductive success was reduced by all oil exposures. Sediment spiked with UCM fractions also caused reduced growth and reproduction but no particular fraction was found to be responsible for the observed toxicity. Survivorship was not affected by any oil or fraction. Discussion  The study showed that chronic exposure to sediments contaminated by UCM-dominated oils could have population level effects on amphipods. The observed effects could not be explained by hydrocarbons resolved by conventional GC and effects were similar for both UCM-dominated and weathered oils. All of the fractions appeared to contribute to the observed effects; this is in contrast to previous research which had shown that an aliphatic UCM did not cause adverse effects in mussels. Conclusions  To our knowledge, this is the first study to demonstrate population-level effects arising from exposure to sediments contaminated by realistic environmental concentrations of UCM hydrocarbons. The results are consistent with many compounds, at very low individual concentrations, contributing towards the overall observed toxicity. Recommendations  Risk assessments of contaminated sediments should take into account the contribution towards the potential for toxic effects from UCM hydrocarbons. Studies into sediment contamination should report both aliphatic and aromatic UCM concentrations to aid risk assessments. ESS-Submission Editor: Henner Hollert (hollert@uni-heidelberg.de) This paper has been developed from a presentation at SETAC Europe 16^th Annual Meeting 2006 held at the The Hague, The Netherlands.     

Microsatellite diversity and effective population size in a germplasm bank of <Emphasis Type="Italic">Hymenaea courbaril</Emphasis> var. <Emphasis Type="Italic">stilbocarpa</Emphasis> (Leguminosae), an endangered tropical tree: recommendations for conservation

Deforestation in southeast Brazil has led to the extinction of Hymenaea courbaril var. stilbocarpa and ex situ conservation has been established. In this study, the levels of genetic diversity and the effective population size of H. courbaril in a germplasm bank were investigated using six nuclear microsatellite loci. A total of 79 and 91 alleles were found in 65 seed-trees and their 176 offspring, respectively. Offspring have a higher average number of alleles per locus (A = 15.2) than seed-trees (A = 13.2), but lower observed heterozygosity (offspring: H _o = 0.566; seed-trees: H _o = 0.607). The estimate of outcrossing rate shows that the study population is perfectly outcrossed (t _m  = 0.978, P > 0.05). Significant deviations from random mating were detected through mating among relatives and correlated matings. The average variance in effective population size for each family was 2.63, with a total effective population size retained in the bank of 170.1. These results confirm that the preserved population of H. courbaril retains substantial genetic variability. Moacyr Antonio Mestriner and Ana Lilia Alzate-Marin—CEEFLORUSP Members.     

Plant canopy effects on litter accumulation and soil microbial biomass in two temperate forests

The objective of this study was to determine whether differences in canopy structure and litter composition affect soil characteristics and microbial activity in oak versus mixed fir-beech stands. Mean litter biomass was greater in mixed fir-beech stands (51.9t ha⁻¹) compared to oak stands (15.7t ha⁻¹). Canopy leaf area was also significantly larger in mixed stands (1.96m² m⁻²) than in oak stands (1.73m² m⁻²). Soil organic carbon (C _org) and moisture were greater in mixed fir-beech stands, probably as a result of increased cover. Soil microbial biomass carbon (C _mic), nitrogen (N _mic), and total soil nitrogen (N _tot) increased slightly in the mixed stand, although this difference was not significant. Overall, mixed stands showed a higher mean C _org/N _tot ratio (22.73) compared to oak stands (16.39), indicating relatively low rate of carbon mineralization. In addition, the percentage of organic C present as C _mic in the surface soil decreased from 3.17% in the oak stand to 2.26% in the mixed stand, suggesting that fir-beech litter may be less suitable as a microbial substrate than oak litter.     

Dinitrogen fixation by biological soil crusts in an Inner Mongolian steppe

Eurasian steppe ecosystems are nitrogen-limited and suffer additionally from high grazing intensities in many areas. Soil surface-bound cyanobacteria are able to fix nitrogen and can be the major source of plant available nitrogen in such ecosystems. In this study, the abundance and dinitrogen fixation capacity of the most common soil surface-bound microbial and lichen species were determined at an ungrazed, a winter-grazed, and a heavily grazed steppe site in the Xilin River catchment, Inner Mongolia, People’s Republic of China. The microorganisms were identified as Nostoc spec. and the lichen species as Xanthoparmelia camtschadalis (Ach.) Hale by a combination of classical light microscopy, confocal laser scanning microscopy and molecular analysis of the internal transcribed spacer (ITS1) region of ribosomal RNA. Both species were found exclusively at grazed steppe sites, with a clear difference in abundance depending on the grazing intensity. At the winter-grazed site, Nostoc was more abundant than Xanthoparmelia; for the heavily grazed site, the opposite was found. N₂ fixation was quantified with both the acetylene reduction method and ¹⁵N₂ incubation. Cyanobacterial colonies of Nostoc fixed N₂ vigorously, whereas X. camtschadalis did not at all. The fraction of nitrogen derived from the fixation of molecular nitrogen in Nostoc was 73%, calculated from ¹⁵N natural abundance measurements of Nostoc with X. camtschadalis as reference. The conservatively calculated N₂ uptake by Nostoc was 0.030–0.033 kg N ha⁻¹ for the heavily grazed site and 0.080–0.087 kg N ha⁻¹ for the winter-grazed site for the growing seasons of 2004 and 2005, respectively. Together with previous findings, this study demonstrates that N₂ fixation by Nostoc can potentially replace significant amounts, if not all, of the nitrogen lost in the form of N₂O and NO soil emissions in this steppe ecosystem.     

Interpreting the dependence of soil respiration on soil temperature and water content in a boreal aspen stand

Continuous half-hourly measurements of soil CO₂ efflux made between January and December 2001 in a mature trembling aspen stand located at the southern edge of the boreal forest in Canada were used to investigate the seasonal and diurnal dependence of soil respiration (R_s) on soil temperature (T_s) and water content (θ). Daily mean R_s varied from a minimum of 0.1 μmol m⁻² s⁻¹ in February to a maximum of 9.2 μmol m⁻² s⁻¹ in mid-July. Daily mean T_s at the 2-cm depth was the primary variable accounting for the temporal variation of R_s and no differences between Arrhenius and Q₁₀ response functions were found to describe the seasonal relationship. R_s at 10 °C (R_s10) and the temperature sensitivity of R_s (Q_10Rs) calculated at the seasonal time scale were 3.8 μmol m⁻² s⁻¹ and 3.8, respectively. Temperature normalization of daily mean R_s (R_sN) revealed that θ in the 0–15 cm soil layer was the secondary variable accounting for the temporal variation of R_s during the growing season. Daily R_sN showed two distinctive phases with respect to soil water field capacity in the 0–15 cm layer (θ_fc, 0.30 m³ m⁻³): (1) R_sN was strongly reduced when θ decreased below θ_fc, which reflected a reduction in microbial decomposition, and (2) R_sN slightly decreased when θ increased above θ_fc, which reflected a restriction of CO₂ or O₂ transport in the soil profile.Diurnal variations of half-hourly R_s were usually out of phase with T_s at the 2-cm depth, which resulted in strong diurnal hysteresis between the two variables. Daily nighttime R_s10 and Q_10Rs parameters calculated from half-hourly nighttime measurements of R_s and T_s at the 2-cm depth (when there was steady cooling of the soil) varied greatly during the growing season and ranged from 6.8 to 1.6 μmol m⁻² s⁻¹ and 5.5 to 1.3, respectively. On average, daily nighttime R_s10 (4.5 μmol m⁻² s⁻¹) and Q_10Rs (2.8) were higher and lower, respectively, than the values obtained from the seasonal relationship. Seasonal variations of these daily parameters were highly correlated with variations of θ in the 0–15 cm soil layer, with a tendency of low R_s10 and Q_10Rs values at low θ. Overall, the use of seasonal R_s10 and Q_10Rs parameters led to an overestimation of daily ranges of half-hourly R_s (ΔR_s) during drought conditions, which supported findings that the short-term temperature sensitivity of R_s was lower during periods of low θ. The use of daily nighttime R_s10 and Q_10Rs parameters greatly helped at simulating ΔR_s during these periods but did not improve the estimation of half-hourly R_s throughout the year as it could not account for the diurnal hysteresis effect.     

Phosphorus release during decomposition of crop residues under conventional and zero tillage

Field experiments were conducted at Fort Vermilion (58°23′N 116°02′W), Alberta, to determine phosphorus (P) release patterns from red clover (Trifolium pratense) green manure (GM), field pea (Pisum sativum), canola (Brassica rapa) and monoculture wheat (Triticum aestivum) residues in the 7th and 8th years of conventional and zero tillage. Phosphorus contained in crop residues ranged from 1.5 kg ha⁻¹ in pea to 9.2 kg ha⁻¹ in clover GM, both under zero tillage. The patterns of P release over a 52-week period sometimes varied with tillage, i.e., a greater percentage of GM residue P was released under conventional tillage than under zero tillage in the first 2–10 weeks of residue placement. Wheat residues resulted in net P immobilization under zero tillage, but the amounts immobilized were less than 1 kg ha⁻¹. When net P mineralization occurred, the percentage of P released ranged from 24% of wheat P under conventional tillage to 74% of GM P under conventional tillage. The amounts of P released were 0.4 kg ha⁻¹ from wheat, 0.8 kg ha⁻¹ from canola, 0.4 kg ha⁻¹ from pea and 5.1–5.6 kg ha⁻¹ from clover GM residues. Therefore, only GM residues recycled agronomically significant amounts of P for use by subsequent crops in rotation. Phosphorus release was positively correlated with residue P concentration and negatively correlated with C/P and lignin/P ratios.     

黔中喀斯特山区坡耕地产流产沙对无籽刺梨种植的响应

产流产沙是坡耕地土壤侵蚀的主要形式,为探讨无籽刺梨对黔中喀斯特山区坡耕地土壤侵蚀治理效应,采用径流小区野外定位观测法和相关性分析法,分析了坡度15°和25°坡耕地共6个自然坡面径流小区2016年6—9月15次野外实测侵蚀性降雨产流产沙数据。结果表明:(1)在同坡度同降雨量同I₃₀下,无籽刺梨地较自然恢复地有显著保水保沙能力,无籽刺梨+自然荒草地模式优于无籽刺梨单种模式。(2)无籽刺梨种植模式减流减沙效果坡度25°优于15°,减沙效果优于减流效果。其中无籽刺梨+自然荒草地减流减沙效果最优,在小雨、中雨、大雨、暴雨4种雨型下,坡度15°度时较自然恢复地分别平均减流59.08%,65.23%,56.96%,54.87%,分别平均减沙87.32%,71.64%,51.10%,74.69%;坡度25°度时分别平均减流53.11%,76.09%,76.72%,46.12%,分别平均减沙85.40%,86.87%,71.55%,73.99%。(3)研究区产流产沙量与降雨参数(P、I₃₀)高度相关且在0.01水平上显著,在不同雨型下无籽刺梨减流减沙均有明显效果,小雨、中雨时减流减沙效果最佳。研究证实了研究区无籽刺梨种植能抵御不同雨型的降雨侵蚀,可作为贵州省喀斯特山区生态修复的极优经济选种。     

Characterization of Swine Wastewater by Toxicity Identification Evaluation Methodology (TIE)

Since swine wastewater is used by farmers for soil fertilization, evaluation of toxic compounds or micro-contaminants of separate streams is required. This paper uses the toxicity identification evaluation (TIE) procedure for the physicochemical and ecotoxicological characterization of swine wastewater. To distinguish the most important toxic compounds, a physicochemical characterization and phase I-TIE procedure were performed. The acute toxic effect of swine wastewater and treated fractions (phase II-TIE) were evaluated using Daphnia magna determining 48-h LC₅₀. Results show a high level of conductivity (23.5 μS cm⁻¹), which is explained as due to the concentration of ions, such as ammonium (NH₄⁺–N 1.6 g L⁻¹), sulfate (SO₄²⁻ 397.3 mg L⁻¹), and chlorine (Cl⁻ 1,230.0 mg L⁻¹). The acute toxicity of the swine wastewater was evaluated on D. magna (48-h LC₅₀ = 3.4%). Results of the different water treatments indicate that anionic exchange treatments could reduce 22.5% of swine wastewater’s acute toxicity by reducing chlorine (to around 51%) and conductivity (8.5%). On the other hand, cationic exchange treatment increased acute toxicity on D. magna (% RT = −624.4%), by reducing NH₄⁺–N (around 100%) and total nitrogen (95.5%). This finding suggests that part of the toxicity comes from anionic compounds, such as chlorine.     

Evaluation of rice–legume–rice cropping system on grain yield,nutrient uptake,nitrogen fixation,and chemical,physical, and biological properties of soil

To achieve higher yields and better soil quality under rice–legume–rice (RLR) rotation in a rainfed production system, we formulated integrated nutrient management (INM) comprised of Azospirillum (Azo), Rhizobium (Rh), and phosphate-solubilizing bacteria (PSB) with phosphate rock (PR), compost, and muriate of potash (MOP). Performance of bacterial bioinoculants was evaluated by determining grain yield, nitrogenase activity, uptake and balance of N, P, and Zn, changes in water stability and distribution of soil aggregates, soil organic C and pH, fungal/bacterial biomass C ratio, casting activities of earthworms, and bacterial community composition using denaturing gradient gel electrophoresis (DGGE) fingerprinting. The performance comparison was made against the prevailing farmers’ nutrient management practices [N/P₂O₅/K₂O at 40:20:20 kg ha⁻¹ for rice and 20:30:20 kg ha⁻¹ for legume as urea/single super-phosphate/MOP (urea/SSP/MOP)]. Cumulative grain yields of crops increased by 7–16% per RLR rotation and removal of N and P by six crops of 2 years rotation increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots over that in compost alone or urea/SSP/MOP plots. Apparent loss of soil total N and P at 0–15 cm soil depth was minimum and apparent N gain at 15–30 cm depth was maximum in Azo/Rh plus PSB dual INM plots. Zinc uptake by rice crop and diethylenetriaminepentaacetate-extractable Zn content in soil increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots compared to other nutrient management plots. Total organic C content in soil declined at 0–15 cm depth and increased at 15–30 cm depth in all nutrient management plots after a 2-year crop cycle; however, bacterial bioinoculants-based INM plots showed minimum loss and maximum gain of total organic C content in the corresponding soil depths. Water-stable aggregation and distribution of soil aggregates in 53–250- and 250–2,000 μm classes increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots compared to other nutrient management plots. Fungal/bacterial biomass C ratio seems to be a more reliable indicator of C and N dynamics in acidic soils than total microbial biomass C. Compost alone or Azo/Rh plus PSB dual INM plots showed significantly (P < 0.05) higher numbers of earthworms’ casts compared to urea/SSP/MOP alone and bacterial bioinoculants with urea or SSP-applied plots. Hierarchical cluster analysis based on similarity matrix of DGGE profiles revealed changes in bacterial community composition in soils due to differences in nutrient management, and these changes were seen to occur according to the states of C and N dynamics in acidic soil under RLR rotation.     

养殖水中总有机碳与化学需氧量和生化需氧量相关性的研究

总有机碳能较全面地反映水体有机物的污染程度,总有机碳的测定具有操作简单、分析快速、结果准确等特点。研究了养殖水中的总有机碳与化学需氧量和生化需氧量相关性,总有机碳与化学需氧量的相关系数为０.９９１８,总有机碳与生化需氧量的相关系数为０.９３６２,可以用总有机碳来估算和衡量水体的污染情况。