Use of agro-industrial organic sludge amendment to remediate degraded soil: chemical and eco(geno)toxicological differences between fresh and stabilized sludge and establishment of application rates

Soil degraded by coal mining activities can be remediated by amendment with agro-industrial organic sludge. However, the environmental impacts associated with this management practice must be properly addressed. In this context, the objective of this study was to evaluate the eco(geno)toxicity of a fresh and a stabilized sludge before use in a laboratory soil remediation test. Chemical analysis of the complex mixtures (degraded soil, fresh sludge, and stabilized sludge) was carried out, as well as a battery of eco(geno)toxicity tests on microbiological enzymes (fluorescein hydrolysis), earthworms, and higher plants (including Vicia faba genotoxicity test), according to published methodologies. The results of these tests showed that fresh sludge was more toxic than sludge stabilized over 6 months toward earthworms and higher plants (lettuce, corn, and wild cabbage), while phyto(geno)toxicity tests with V. faba indicated the same genotoxicity levels for the two types of sludge. In the soil remediation simulation using different mixtures of degraded soil and stabilized sludge, the proportions of 50:50 % (dry weight basis) provided the lowest phyto(geno)toxicity effects and this mixture can be used for the revegetation of the contaminated site.     

Leachates from solid wastes: chemical and eco(geno)toxicological differences between leachates obtained from fresh and stabilized industrial organic sludge

The chemical and ecotoxicological characteristics of fresh and stabilized industrial organic sludge leachates were compared to obtain information regarding how the stabilization process can influence the ecotoxic potential of this industrial waste, which could be used for the amendment of degraded soil. Physicochemical analysis of the sludge leachates, as well as a battery of eco(geno)toxicity tests on bacteria, algae, daphnids, and higher plants (including Vicia faba genotoxicity test) and the determination of hydrolytic enzyme activity, was performed according to standard methods. The chemical comparison of the two types of leachate showed that the samples obtained from stabilized sludge had a lower organic content and higher metal content than leachates of the fresh sludge. The eco(geno)toxicological results obtained with aquatic organisms showed that the stabilized sludge leachate was more toxic than the fresh sludge leachate, both originating from the same industrial organic sludge sample. Nevertheless, phytotoxicity tests carried out with a reference peat soil irrigated with stabilized sludge leachate showed the same toxicity as the fresh sludge leachate. In the case of the industrial solid organic sludge studied, stabilization through a biodegradation process promoted a higher metal mobility/bioavailability/eco(geno)toxicity in the stabilized sludge leachate compared to the fresh sludge leachate.     

Metal availability and uptake by sorghum plants grown in soils amended with sludge from different treatments

Several factors depending on the sludge, the soil, or the combination of both substrates, may affect element availability to plants. In this study, an assessment was done of the effect of two sludges obtained by different processes (activated sludge and facultative stabilization pond) on heavy-metal availability and uptake by sorghum plants in soils with high and low copper contents. Results obtained for DTPA-extractable metal indicated higher metal availability in sludge-amended soils. In addition, sludges caused changes in copper and zinc distribution in soil, indicating in most cases a discrete increase in the more labile metal forms. However, observed changes did not increase heavy metal concentration in plant leaves, indicating that assessment of metal availability by a chemical procedure (single extraction or metal fractionation) would not permit a good prediction of metal bioavailability. On the other hand, sludge application at a rate of 100 t ha⁻¹ to high-copper agricultural soils would not imply greater mobility of this metal on account of a greater sorbing capacity provided by the sludges. Such results would indicate that sludges from wastewater treatment plants, meeting the standards of heavy metal contents, regardless of the process by which they were obtained, may be applied to several kinds of soil, even to high-copper soils, with no risk of increasing heavy metal bioavailability to phytotoxic levels in the short range.     

Functional and community-level soil microbial responses to zinc addition may depend on test system biocomplexity

The effect of zinc on soil nitrification and composition of the microbial community in soil was investigated using a full factorial experiment with five zinc concentrations and four levels of biological complexity (microbes only, microbes and earthworms (Eisenia fetida), microbes and Italian ryegrass (Lolium multiflorum var. Macho), and microbes, ryegrass and earthworms). After 6 weeks of exposure, the activity of soil nitrifying bacteria was measured and the microbial community structure was characterized by phospholipid fatty acid (PLFA) analysis. Soil nitrification and several PLFA markers were significantly influenced by either zinc addition and/or the presence of earthworms or ryegrass, and one of the most pronounced changes was the increase of fungi and decrease of bacteria with increasing concentrations of zinc. Of particular interest, however, was the potential interaction between the presence of plants and/or earthworms and the effect of zinc, which the factorial study design allowed us to explore. Such an effect was observed in two cases: Earthworms reduced the positive effect of zinc on the fungal biomass (ANOVA, p=0.03), and the effect of earthworms on the soil nitrification activity depended on zinc concentration (ANOVA, p<0.05). The effect of earthworm presence was not very large, but it does show that multispecies tests might give information about metal toxicity or bioavailability that cannot be predicted from single-species tests.     

Optimization of animal manure vermicomposting based on biomass production of earthworms and higher plants

The goal of this study was to optimize the mixture of swine manure (SM) and cattle manure (CM) used in the vermicomposting process, seeking to increase the manure biodegradation rate and enhance the biomass production of both earthworms and higher plants. To achieve this goal, physico-chemical parameters were determined to assess the final compost quality after 50 days of vermicomposting. The different manure ratios used to produce the composts (C) were as follows (SM:CM, % m/m basis): C1 100:0, C2 (75:25), C3 (50:50), C4 (25:75), and C5 (0:100). In addition, the earthworm biomass and the phytoproductivity of lettuce (Lactuca sativa L.) plants grown in mixtures (1:1) of natural soil and the most viable vermicomposts were investigated. The C1 and C2 compost compositions were associated with high earthworm mortality rates. The C3 compost provided the highest mineral concentrations and C5 showed the highest lettuce yield (wet biomass). The results verify that stabilized cattle manure is an excellent substrate for the vermicomposting process and that fresh swine manure must be mixed with pre-stabilized cattle manure to ensure an optimized vermicomposting process, which must be controlled in terms of temperature and ammonia levels. It is concluded that small livestock farmers could add value to swine manure by applying the vermicomposting process, without the need for high investments and with a minimal requirement for management of the biodegradation process. These are important technical aspects to be considered when circular economy principles are applied to small farms.     

Comparison of sewage sludge toxicity to plants and invertebrates in three different soils

Understanding the effect of soil type on the overall toxicity of sewage sludge is one of the most important issues concerning environmental risks associated with the sewage sludge land application. The aim of the study was to determine the influence of different soils (sandy, loamy and OECD soil) on sewage sludges toxicity in relation to plants (Lepidium sativum, Sorghum saccharatum, Sinapis alba) and an invertebrate species (Heterocypris incongruens). The most evident negative influence of sewage sludges on root growth was observed in the case of OECD soil. The EC(50) values determined on the basis of the root growth inhibition of all tested plants were in the range 0.1-6.4%, 0.03-9.4% and 6.6-22.1% (% of sewage sludgekg(-1) soil) for OECD, sandy and loamy soil, respectively. Soil type also affects the sewage sludge toxicity in relation to H. incongruens. The LC(50) (mortality) values ranged from 0.26% to 11.5% depending on the sludge tested. For EC(50) (growth inhibition) values ranged from 10.7% to 36.2%.     

Acute toxicity of chlorophenols to earthworms using a simple paper contact method and comparison with toxicities to fresh water organisms

An acute toxicity test of chlorophenols on earthworms (Eisenia fetida) was performed using a simple paper contact method proposed by OECD testing guideline no. 207, that were applied as an earthworm toxicity test. The median lethal concentration, EC50, had significant correlation with logP(ow) (1-octanol/water partition coefficient) of the chemicals. The toxicity of chlorophenols on E. fetida was compared with toxicities for other species: an algae (Selenostrum capricornutum), a crustacean (Daphnia magna), and a fish (Oryzias latipes). It was found that the toxicity of chlorophenols was almost same for E. fetida and for fresh water organisms. These results suggest the possibility of drawing correlations between the effects of pollutants on living things in different environments, fresh water and soil.     

Reduction of bioavailability and leachability of heavy metals during vermicomposting of water hyacinth

Vermicomposting of water hyacinth is a good alternative for the treatment of water hyacinth (Eichhornia crassipes) and subsequentially, beneficial for agriculture purposes. The bioavailability and leachability of heavy metals (Zn, Cu, Mn, Fe, Ni, Pb, Cd, and Cr) were evaluated during vermicomposting of E. crassipes employing Eisenia fetida earthworm. Five different proportions (trials 1, 2, 3, 4, and 5) of cattle manure, water hyacinth, and sawdust were prepared for the vermicomposting process. Results show that very poor biomass growth of earthworms was observed in the highest proportion of water hyacinth (trial 1). The water soluble, diethylenetriaminepentaacetic acid (DTPA) extractable, and leachable heavy metals concentration (percentage of total heavy metals) were reduced significantly in all trials except trial 1. The total concentration of some metals was low but its water soluble and DTPA extractable fractions were similar or more than other metals which were present in higher concentration. This study revealed that the toxicity of metals depends on bioavailable fraction rather than total metal concentration. Bioavailable fraction of metals may be toxic for plants and soil microorganisms. The vermicomposting of water hyacinth by E. fetida was very effective for reduction of bioavailability and leachability of selected heavy metals. Leachability test confirmed that prepared vermicompost is not hazardous for soil, plants, and human health. The feasibility of earthworms to mitigate the metal toxicity and to enhance the nutrient profile in water hyacinth vermicompost might be useful in sustainable land renovation practices at low-input basis.     

Abandoned coal mining sites: using ecotoxicological tests to support an industrial organic sludge amendment

The different stages involved in coal mining-related activities result in a degraded landscape and sites associated with large amounts of dumped waste material. Remediation of these contaminated soils can be carried out by application of industrial organic sludge if the concerns regarding the potential negative environmental impacts of this experimental practice are properly addressed. In this context, the objective of this study was to use ecotoxicological tests to determine the quantity of organic industrial sludge that is required as a soil amendment to restore soil production while avoiding environmental impact. Chemical analysis of the solids (industrial sludge and soil) and their leachates was carried out as well as a battery of ecotoxicity tests on enzymes (hydrolytic activity), bacteria, algae, daphnids, earthworms, and higher plants, according to standardized methodologies. Solid and leachate samples of coal-contaminated soil were more toxic than those of industrial sludge towards enzyme activity, bacteria, algae, daphnids, and earthworms. In the case of the higher plants (lettuce, corn, wild cabbage, and Surinam cherry) the industrial sludge was more toxic than the coal-contaminated soil, and a soil/sludge mixture (66:34 % dry weight basis) had a stimulatory effect on the Surinam cherry biomass. The ecotoxicological assessment of the coal-contaminated soil remediation using sludge as an amendment is very important to determine application rates that could promote a stimulatory effect on agronomic species without negatively affecting the environment.     

Assessment of the toxic potential of hydrocarbon containing sludges

A short-time period microbial toxicity test-battery was used for the investigation of acute toxicity and genotoxicity of five hydrocarbon containing sludges. Four sludges were obtained from a petrochemical industry and the fifth from a petroleum refinery. Some of the sludges had been stored for long periods. Bioremediation potential assays for soils polluted with each of the sludges were also considered. The sludges did not show acute toxicity in any of the microbial tests performed. However, when the diethylether soluble fractions of these sludges were analyzed some of them showed acute toxicity, for which the clearest results were obtained with the resazurin reduction method. The greatest toxicity detected with the Resazurin based method was found in the diethylether extracts of the freshly collected (not stored) sludges. On the other hand, the diethylether soluble fraction of those sludges that had been stored showed genotoxicity when analyzed with the Salmonella/microsome assay. After the incorporation of the sludges into the soil, increased bacterial counts were noted and substantial hydrocarbon elimination was achieved in 30 days, showing that bioremediation may be a possible technology for cleaning soils polluted with these sludges.     

Overcoming the toxicity effects of municipal wastewater sludge and biosolid extracts in the Yeast Estrogen Screen (YES) assay

For nearly two decades, the Yeast Estrogen Screen (YES) has been used as a valuable tool for determining the total estrogenic potency of various environmental samples, including influent and effluent streams at municipal wastewater plants. However, applying the YES assay to wastewater sludges and stabilized biosolids has been problematic. This is due to co-extracted compounds from the solids either proving toxic to the yeast or masking the presence of estrogenic substances. The present research describes the development and validation of sample preparation steps that mitigate the toxicity effects of municipal wastewater sludge and biosolid samples in the YES assay, while allowing for reliable dose-dependent expression of estrogenic activity. A copper work-up for sulfur removal and chromatographic cleanup with silica and alumina were required in addition to solid-phase extraction to adequately remove interfering compounds. Sample stabilization methods such as autoclaving, lyophilization and formaldehyde treatment were found to be detrimental to the assay. Hence, heat-drying is recommended to prevent cytotoxicity and the degradation of estrogenic substances.     

Leaching and reduction of chromium in soil as affected by soil organic content and plants

The oxidation state of chromium in contaminated soils is an important indicator of toxicity and potential mobility. Chromium in the hexavalent state is highly toxic and soluble, whereas the trivalent state is much less toxic and relatively insoluble. A laboratory study investigated the impact of growing plants and supplemental organic matter on chromium transport in soil. Plants alone had no appreciable effect on the chromium oxidation state in soil. Soil columns with higher organic content were associated with lower ratios of chromate:total chromium than the columns with lower organic matter. Analyses of column leachate, plant biomass, and soil indicate that more chromium leaching occurred in the vegetated, low organic columns. Retention of Cr in the soils was correlated to the Cr(III) content. Plant uptake of chromium accounted for less than 1% of the chromium removed from the soil. Overall, the addition of organic matter had the strongest influence on chromium mobility.     

Influence of soil properties on bioaccumulation of 14C-simazine in earthworms Eisenia foetida

The toxicity of pesticides has been evaluated by several methods including tests with earthworms in both artificial and natural soils treated with the compounds. The ecological niches of earthworms make them good bioindicators of soil contamination. The bioaccumulation of 14C-simazine (6-chloro-N2-N4-diethyl- 1,3,5-triazine-2,4-diamine) was evaluated in earthworms (Eisenia foetida) maintained during three months in two substrates with different physical-chemical characteristics. The substrates were treated with 3.0 mg and 330 kBq of 14C-simazine kg(-1) substrate. Results indicated that worms did not influence simazine dissipation in both substrates as indicated by similar recoveries and with no statistical differences with and without earthworms. The radiocarbon recoveries were 86.8 and 95.3%, respectively in the substrates with lower and higher organic matter contents with earthworms, and 91.0 and 107.4% in the same substrates without worms. However, in earthworms the recoveries were statistically higher when they were maintained in the substrate with lower amount of organic matter (0.89%) than from the higher one (0.33%). Consequently, 14C-simazine bioconcentration factor (BCF) was also greater in the substrate with lower organic matter (6.89+/-1.55) than in the substrate with higher organic matter content (0.88+/-0.06). The results suggest that the higher soil organic matter content will cause lower probability of contamination of soil organisms with simazine.     

Effect of C/N ratios on the performance of earthworm eco-filter for treatment of synthetics domestic sewage

Purpose

The performances of filter systems that use earthworms and plants, combined with earthworm eco-filter (EE) systems in treating synthetic domestic sewage (SDS) with different C/N ratios, were investigated for a 9-month period.

Methods

The effects of the combination of filters, earthworms, plants, as well as the combination of earthworms and plants on SDS nutrient removal efficiency were separately investigated to select the optimum system for treating SDS. The results of the current study could be used to determine how treatment performance responds to different C/N ratios and to explain and predict the performance of an operating EE system.

Results

EE systems with earthworms and plants (EP groups) consistently performed better than the other types of systems (CK, E, and P; that is, without earthworms and without plants, with earthworms and without plants, and without earthworms and with plants, respectively) under all C/N ratios. The highest removal efficiencies of chemical oxygen demand, total nitrogen, total phosphorus, and total organic carbon were achieved under C/N ratios of 6:1, 6:1, 6:1, and 9:1, respectively. The optimum nutrient removal efficiency was achieved at C/N?=?6, and the contribution order for nutrient removal was EP > P > E > CK.

Conclusions

Influent C/N ratios, the time of year, and the synergetic effects of earthworm behavior and microorganisms significantly affected nutrient removal efficiencies. Considering the removal of all nutrients, EE systems with plants and earthworms achieved optimum removal effects in July when the influent C/N ratio was controlled at 6. Appropriate control of carbon and nitrogen source concentrations permitted the achievement of optimal nutrient removal effects.     

Effects of spatial and temporal variation in metal availability on earthworms in floodplain soils of the river Dommel, The Netherlands

Regarding impact on ecological soil functioning, metal pollution is often considered a constant factor for certain sampling sites. However, especially bioavailable concentrations may differ in space and time. This aspect was investigated on four sites along a metal-polluted river, differing in soil characteristics and metal concentrations. Every four weeks earthworm densities, soil characteristics, and metal concentrations in soil and earthworms were determined. Earthworm biomass and density fluctuated in time and increased with increasing metal contamination, indicating the presence of compensating factors. Multivariate analysis suggested organic matter and moisture content to be the main factors explaining earthworm biomass. Metal concentrations in the earthworms increased with increasing total or 0.01M CaCl(2) extractable soil concentrations, but no time-related trends were seen. Cadmium concentrations in the earthworms exceeded background values, suggesting a potential risk. The neutral red retention biomarker assay, however, did not show any signs of metal stress in the earthworms.     

Arbuscular mycorrhizal fungi mediated uptake of lanthanum in Chinese milk vetch (Astragalus sinicus L.)

A glasshouse experiment was carried out to study the effect of mycorrhizal formation by Gigaspora margarita, Glomus intraradices or Acaulospora laevis on plant growth and lanthanum (La) uptake of Astragalus sinicus L. in soils spiked with La at five levels (0, 1, 5, 10 and 20 mg kg(-1)). La application decreased mycorrhizal infection frequency and activity of fungal succinate dehydrogenase and alkaline phosphatase. Increasing La concentrations in soil led to increased La accumulation in tissues of both mycorrhizal and non-mycorrhizal plants, but inoculation with Gig. margarita or G. intraradices reduced La concentrations in shoots and roots at higher concentrations of La in the soil. Plant biomass and P concentrations in shoots and roots were reduced by La application, but increased by inoculation with Gig. margarita or G. intraradices. The results showed that appropriate AM fungi colonization could be effective in alleviating La toxicity in A. sinicus.     

Effects of phenanthrene on the mortality, growth, and anti-oxidant system of earthworms (Eisenia fetida) under laboratory conditions

To assess the toxic effects of phenanthrene on earthworms, we exposed Eisenia fetida to artificial soils supplemented with different concentrations (0.5, 2.5, 12.5, mgkg(-1) soil) of phenanthrene. The residual phenanthrene in the soil, the bioaccumulation of phenanthrene in earthworms, and the subsequent effects of phenanthrene on growth, anti-oxidant enzyme activities, and lipid peroxidation (LPO) were determined. The degradation rate of low concentrations of phenanthrene was faster than it was for higher concentrations, and the degradation half-life was 7.3d (0.5 mgkg(-1)). Bioaccumulation of phenanthrene in the earthworms decreased the phenanthrene concentration in soils, and phenanthrene content in the earthworms significantly increased with increasing initial soil concentrations. Phenanthrene had a significant effect on E. fetida growth, and the 14-d LC(50) was calculated as 40.67 mgkg(-1). Statistical analysis of the growth inhibition rate showed that the concentration and duration of exposure had significant effects on growth inhibition (p<0.001). Superoxide dismutase (SOD) activity increased at the beginning (2 and 7d) and decreased in the end (14 and 28 d). Catalase (CAT) activity in all treatments was inhibited from 1 to 14 d of exposure. However, no significant perturbations in malondialdehyde (MDA) content were noted between control and phenanthrene-treated earthworms except after 2d of exposure. These results revealed that bioaccumulation of phenanthrene in E. fetida caused concentration-dependent, sub-lethal toxicity. Growth and superoxide dismutase activity can be regarded as sensitive parameters for evaluating the toxicity of phenanthrene to earthworms.     

Assessing the chemical and biological accessibility of the herbicide isoproturon in soil amended with biochar

Decamethylcyclopentasiloxane (D5) is a cyclic volatile methyl siloxane (cVMS) commonly found in commercially available products. D5 is expected to enter the terrestrial environment through the deposit of biosolids from sewage treatment plants onto agricultural fields for nutrient enrichment. Little to no information currently exists as to the risks of D5 to the terrestrial environment. In order to evaluate the potential risk to terrestrial organisms, the toxicity of a D5 contaminated biosolid in an agricultural soil was assessed with a battery of standardized soil toxicity tests.D5 was spiked into a surrogate biosolid and then mixed with a sandy loam soil to create test concentrations ranging from 0 to 4074 mg kg⁻¹. Plant (Hordeum vulgare (barley) and Trifolium pratense (red clover)) and soil invertebrates (Eisenia andrei (earthworm) and Folsomia candida (springtail)) toxicity tests were completed to assess for lethal and sub-lethal effects. Plant testing evaluated the effects on seedling emergence, shoot and root length, and shoot and root dry mass. Invertebrate test endpoints included adult lethality, juvenile production, and individual juvenile dry mass (earthworms only). Soil samples were collected over time to confirm test concentrations and evaluate the loss of chemical over the duration of a test. The toxicity of the D5 was species and endpoint dependent, such that no significant adverse effects were observed for T. pratense or E. andrei test endpoints, however, toxicity was observed for H. vulgare plant growth and F. candida survival and reproduction. Chemical losses of up to 50% were observed throughout the tests, most significantly at high concentrations.     

Calcium-lead interactions involving earthworms. Part 1: The effect of exogenous calcium on lead accumulation by earthworms under field and laboratory conditions

Earthworms (Lumbricus rubellus and Dendrodrilus rubidus) were collected from several acidic and calcareous abandoned ferrous metalliferous mine sites. Tissue lead concentrations were substantially lower than the total soil lead concentrations, except at one site (Cwmystwyth) where the tissue lead concentrations of both species were approximately 5 to 10 times higher than that of the soil. Soil lead was the major factor in determining the tissue lead concentration, although it was demonstrated that both soil pH and soil calcium concentration could markedly increase the % variance in tissue lead concentration. These findings help explain the apparent anomaly in tissue lead concentrations of earthworms from Cwmystwyth, where the soil is acidic and has exceptionally low calcium concentrations. Soil-liming experiments provided supportive evidence that soil pH, coupled with soil calcium, influences lead accumulation by earthworms, but a filter paper feeding experiment provided unequivocal evidence that soil calcium concentration alone can influence lead accumulation by earthworms. It is concluded that, although lead accumulation by earthworms is influenced by both physico-chemical and biochemical mechanisms, the latter over-rides the former, i.e. soil calcium is more important factor in determining the accumulation of lead earthworms than is soil pH.     

Responses of earthworm to aluminum toxicity in latosol

Excess aluminum (Al) in soils due to acid rain leaching is toxic to water resources and harmful to soil organisms and plants. This study investigated adverse impacts of Al levels upon earthworms (Eisenia fetida) from the latosol (acidic red soil). Laboratory experiments were performed to examine the survival and avoidance of earthworms from high Al concentrations and investigate the response of earthworms upon Al toxicity at seven different Al concentrations that ranged from 0 to 300 mg kg^?1 over a 28-day period. Our study showed that the rate of the earthworm survival was 100 % within the first 7 days and decreased as time elapsed, especially for the Al concentrations at 200 and 300 mg kg^?1. A very good linear correlation existed between the earthworm avoidance and the soil Al concentration. There was no Al toxicity to earthworms with the Al concentration ≤50 mg kg^?1, and the toxicity started with the Al concentration ≥100 mg kg^?1. Low Al concentration (i.e., <50 mg kg^?1) enhanced the growth of the earthworms, while high Al concentration (>100 mg kg^?1) retarded the growth of the earthworms. The weight of earthworms and the uptake of Al by earthworms increased with the Al concentrations from 0 to 50 mg kg^?1 and decreased with the Al concentrations from 50 to 300 mg kg^?1. The protein content in the earthworms decreased with the Al concentrations from 0 to 100 mg kg^?1 and increased from 100 to 300 mg kg^?1. In contrast, the catalase (CAT) and superoxide dismutase (SOD) activities in the earthworms increased with the Al concentrations from 0 to 100 mg kg^?1 and decreased from 100 to 300 mg kg^?1. The highest CAT and SOD activities and lowest protein content were found at the Al concentration of 100 mg kg^?1. Results suggest that a high level of Al content in latosol was harmful to earthworms.