Detoxification and eco-friendly recycling of brick kiln coal ash using Eisenia fetida: A clean approach through vermitechnology.

Brick kiln coal ashes (BKCAs) are one of the major toxic byproducts of the rapidly growing construction industry in developing countries. However, eco-friendly recycling avenues for BKCAs are yet to be explored. The major objectives of the present research were to evaluate the viability of vermitechnology in transforming BKCAs into valuable products, and to examine the metal detoxification potential of Eisenia fetida BKCA-based feedstocks. BKCAs were mixed in large scale with cow dung (CD) in 1:1 and 2:1 ratios, for vermicomposting and aerobic composting; performance was assessed in comparison with CD. Vermiconverted-BKCA was then used as organic fertilizer for rice grown in poorly fertile soil. Acidic nature of BKCA feedstocks was neutralized by 30-86% in the vermireactors. Total N and available P concentrations significantly increased in the vermireactors supplemented with considerable mineralization of total organic C. Exorbitantly high K and S contents were pacified to a normal range after vermicomposting. Greater improvement in microbial biomass, respiration, fungal and bacterial growth was observed under vermicomposting against aerobic composting. Consequently, urease and phosphatase activity increased by 1-4 folds in the BKCA based vermibeds. Bioavailability of toxic metals reduced by 41-74% in the vermicomposted BKCAs. High metal accumulation by the earthworms resulted in substantial reduction of pollution load in the finished product. The field experiment demonstrated that vermiconverted-BKCA could be utilized as potential organic fertilizer for rice production, soil fertility rejuvenation, and metal detoxification. Overall, the study reveals that E. fetida could be used as an efficient contender for sanitization of toxic BKCAs.

Promoting Potato Seed Sprouting Using an Amphiphilic Nanocomposite.

Most potato tubers were used as seeds and sprouted relatively slowly in soil, greatly influencing potato production. To solve this problem, an amphiphilic nanocomposite was fabricated by loading hydrophobic silica (H-SiO2) in hydrophilic attapulgite nest-like and used as a nano presprouting agent (NPA). This technology could conveniently adjust the occupation area ratio of water and air (OARWA) on the potato surface. NPA could endow potatoes with an appropriate OARWA and, thus, effectively accelerate sprouting. Additionally, NPA greatly decreased soil bulk density, facilitated earthworm growth, promoted potato growth, and increased the yield by 14.1%. Besides, NPA did not pass through the potato skin and mainly existed on the surface of potatoes. Importantly, NPA showed tiny influence on the viability of fish and nematodes, demonstrating good biosafety. Therefore, this work provides a promising presprouting approach for potatoes, which may have a potential application prospect in ensuring food supply.

Differences in the bioaccumulation of selenium by two earthworm species (Pheretima guillemi and Eisenia fetida).

Information on the bioaccumulation of selenium (Se) in soil invertebrates (e.g. earthworms) is rather scarce. In the present study, bioaccumulation of Se in two eco-physiologically different earthworms, namely anecic Pheretima guillemi and epigeic Eisenia fetida, was determined after 28 days exposure to a successive doses of Se-spiked soil, specifically 0.5, 5, 50, and 200 µg Se g-1 soil. The results showed that Se concentration in earthworms elevated with increasing exposure levels, and maximums were up to 54.6 and 83.0 µg g-1 dry weight in Pheretima guillemi and Eisenia fetida, respectively, after 4 weeks exposure to 200 µg Se g-1 soil. Exposure to Se caused significant inhibition on earthworm growth, with the fresh weight loss ranging from 8.9% to 80.5%. Bioaccumulation factors (BAFs), empirically-derived and non-steady state, ranged from 0.12 to 4.17 and generally declined at higher exposure levels. Moreover, BAFs of Pheretima guillemi were higher than those of Eisenia fetida in low-dose Se-spiked soils, but the opposite was true in high-dose soils, indicating there is a species-specific response to exposure of Se between different earthworms. Further research is thus needed to reveal the accumulation pattern of Se in a wider range of earthworm species other than Eisenia fetida, which allows a better risk assessment of excessive Se to soil invertebrates and higher order organisms.

Distinct effects of the nephridial symbionts Verminephrobacter and Candidatus Nephrothrix on reproduction and maturation of its earthworm host Eisenia andrei.

Verminephrobacter, the most common specific symbionts in the nephridia (excretory organs) of lumbricid earthworms, have been shown to improve reproduction of the garden earthworm Aporrectodea tuberculata under nutrient limitation. It is unknown how general this beneficial trait is in the Verminephrobacter-earthworm symbiosis, whether other nephridial symbionts also affect host fitness and what the mechanism of the fitness increase is. Here we report beneficial effects of Verminephrobacter and Candidatus Nephrothrix on life history traits of the compost worm Eisenia andrei, which in addition to these two symbionts also hosts Agromyces-like bacteria in its mixed nephridial community: while growth was identical between control, Verminephrobacter-free and aposymbiotic worms, control worms produced significantly more cocoons and offspring than both Verminephrobacter-free and aposymbiotic worms, confirming the reproductive benefit of Verminephrobacter in a second host with different ecology and feeding behavior. Furthermore, worms with Verminephrobacter and Ca. Nephrothrix, or with only Ca. Nephrothrix present, reached sexual maturity significantly earlier than aposymbiotic worms; this is the first evidence for a beneficial role of Ca. Nephrothrix in earthworms. Riboflavin content in cocoons and whole earthworms was unaffected by the presence or absence of nephridial symbionts, suggesting that nutritional supplementation with this vitamin does not play a major role in this symbiosis.

Selenium toxicity to survival and reproduction of Collembola and Enchytraeids in a sandy loam soil.

We investigated the toxicity of selenium (Se) to the soil invertebrates Folsomia candida (Collembola) and Enchytraeus crypticus (potworm). Studies were designed to generate ecotoxicological benchmarks for developing ecological soil screening levels (Eco-SSLs) for risk assessments of contaminated soils. For the present studies, we selected Sassafras sandy loam, an aerobic upland soil with soil characteristics (low levels of clay and organic matter, soil pH adjusted from 5.2 to 7.1) that support high relative bioavailability of the anionic Se species that is typically found in aerobic soil. The Se was amended into soil as sodium selenate, subjected to weathering and aging using 21 d of alternating cycles of air-drying/rehydration to 60% of the water-holding capacity of the Sassafras sandy loam soil, under ambient greenhouse conditions. Effective concentrations at 20 and 50% (EC20 and EC50) levels for production of juveniles (reproduction) were 4.7 and 10.9 mg of Se/kg of soil (dry mass basis), respectively, for Collembola, and 4.4 and 6.2 mg/kg, respectively, for the potworms. The data enabled the derivation of toxicity benchmarks, contributing to the development of a soil invertebrate-based Eco-SSL of 4.1 mg/kg for Se. Environ Toxicol Chem 2018;37:846-853. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Toxic effects of Solanum xanthocarpum Sch &Wendle against Helicoverpa armigera (Hub.), Culex quinquefasciatus (Say.) and Eisenia fetida (Savigny, 1826).

Many commercially available agro and household chemicals are used as pesticides, repellents, and growth inhibitors against insect pests. The repeated uses of these chemicals against insect pests have caused the development of resistance in them; they also cause ill effects on nontarget organisms. The present study was aimed to evaluate the antifeedant, larvicidal, pupicidal, and biochemical effects of the solvent extracts of Solanum xanthocarpum against third instar larvae of Helicoverpa armigera. Hexane, chloroform and ethyl acetate extracts were subjected to phytochemical analysis. The results revealed the presence of terpenoids, flavonoid, and quinone. Maximum antifeedant activity of 72.30% was recorded in chloroform extract followed by hexane (69.02%) and ethyl acetate (57.40%) extracts against H. armigera. Chloroform extracts of S. xanthocarpum showed more than 60% larvicidal and pupicidal activity against H. armigera. The effective chloroform extract was fractionated with increasing polarity of solvent system (hexane, chloroform, and ethyl acetate extracts). Based on the TLC profile, nine major fractions were isolated. The fourth fraction showed higher antifeedant, larvicidal, and pupicidal activity against H. armigera. The effective fraction reduced the hemolymph and gut protein concentration in a concentration-dependent manner (r 2 0.99). The effective fraction 4 showed 100% larvicidal activity at 500 ppm concentration with LC50 value of 227.95 ppm. The fourth fraction did not show any toxic symptom or mortality of earthworm. Based on these results, this effective fraction could be used in the development of a pesticide formulation to control insect.

The impact of modified nano-carbon black on the earthworm Eisenia fetida under turfgrass growing conditions: Assessment of survival, biomass, and antioxidant enzymatic activities.

Modified nano-carbon adsorbents have been employed in the immobilization of heavy metals in soil due to their good adsorption capabilities regarding metal ions. However, an assessment of their risks has not been extensively performed with soil organisms. To assess the toxic effects of three types of modified nano-carbon black (CB) on soil organisms, a laboratory test was conducted to expose the earthworm Eisenia fetida to artificial soil supplemented with 5% H2SO4-, HNO3- and KMnO4-modified nano-CB (SCB, NCB and KCB, respectively) under turfgrass growing conditions. The tested earthworms were systematically investigated for survival, biomass and the activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). SCB and NCB were found to be more toxic and ecologically dangerous to E. fetida because significant decreases in biomass and survival were observed after 35- and 60-d exposures and the survival rate showed a tendency to decrease with exposure duration. The activities of SOD, CAT and POD were inhibited in all treatments with modified nano-CBs at 35- and 60-d, which indicated that oxidative stress was induced by modified nano-CBs. The results suggest that there is potential harm to earthworms in soil with 5% modified nano-CB and that it deserves special attention.

Value added product recovery from sludge generated during gum arabic refining process by vermicomposting.

Gum arabic is multifunctional and used in food products, pharmaceutical, confectionery, cosmetic, printing and textile industry. Gum arabic has an excellent market and its production is being increased to meet the market demand. In the process, huge quantity of solid waste is generated during its refining process. An attempt has been made to vermicompost this organic waste using Eudrilus eugeniae. This research work is first of its kind. Literature on this substrate has not been reported anywhere else for vermicomposting. Results were excellent with volatile solid reduction of 51.34 %; C/N ratio reduced to 16.31 % indicating efficient loss of carbon as carbon dioxide during vermicomposting period. Manurial value, i.e. nitrogen, phosphorus and potassium content in the range, required for the plants also increased. Porosity of 67.74 % and water holding capacity of 65.75 % were observed. The maturity of the vermicompost was evaluated through scanning electron microscopy wherein the complete conversion of large raw material particles into finer particles forming a uniform matrix with more surface area was observed indicating its efficient conversion. Microbial quality of vermicompost was also studied. The final vermicompost is free of fungal cells and pathogenic bacteria.

Adaptability comparison of E. fetida in vermicomposting against sludge from livestock wastewater treatment plant based on their several growth stages.

Vermicomposting is a low-cost, eco-efficient process to deal with organic wastes. Mixtures of swine manure (SM), cow dung (CD), and animal wastewater treatment plant sludge (S) were applied as feeds, and Eisenia fetida was employed in this study to investigate the vermicomposting efficiency based on their several growth stages. The hatching test resulted in a 100 % hatching rate in S60SM40 (60 % S + 40 % SM) mixture, while 4.40 hatchlings per cocoon were observed. The growth of infancy performed best in 0-20 % CD mixtures (0.05 ± 0.002 g), followed by in SM + CD (0.04 ± 0.003 g). The highest growth rate of young and adult E. fetida was noticed in CD + S mixtures (11.14 ± 0.01 and 6.00 ± 0.02 mg/d/worm, respectively), while the higher cocoon production of adults was noticed in S + SM mixtures especially in S40SM60 (537 ± 5 worms). Moreover, the conversion of solids; the modified pH value; the reduction in total organic carbon (TOC); total Kjeldahl nitrogen (TKN), NH4-N, NO3-N, and C:N ratio; and the rich in total available phosphorus (TAP) and total potassium (TK) content by young and adult E. fetida were related to the growth of worms. Such work would benefit understanding and to increase the efficiency of vermicompost processing of different wastes.

Influence of stocking density on the vermicomposting of an effluent treatment plant sludge amended with cow dung.

This paper reports the effect of earthworm population density on the vermicomposting of effluent treatment plant sludge of a bakery industry. Four waste mixtures containing 0, 10, 20, and 30 % sludge along with cow dung with five different worm population densities were established for 14 weeks under controlled moisture and temperature conditions. The results showed that average worm biomass, growth and cocoon production were lesser at higher population densities. Sexual maturity was attained in 3rd to 5th week in all waste mixtures. Worm growth was inversely related to worm population density in the waste mixture. Results also indicated that lower worm population is favorable to worm biomass production. On the other hand, mineralization and stabilization of the waste mixtures were more at higher worm populations.

Dynamics of microbiological parameters, enzymatic activities and worm biomass production during vermicomposting of effluent treatment plant sludge of bakery industry.

This paper reports the changes in microbial parameters and enzymatic activities during vermicomposting of effluent treatment plant sludge (ETPS) of bakery industry spiked with cow dung (CD) by Eisenia fetida. Six vermibins containing different ratios of ETPS and CD were maintained under controlled laboratory conditions for 15 weeks. Total bacterial and total fungal count increased upto 7th week and declined afterward in all the bins. Maximum bacterial and fungal count was 31.6 CFU × 10(6) g(-1) and 31 CFU × 10(4) g(-1) in 7th week. Maximum dehydrogenase activity was 1921 µg TPF g(-1) h(-1) in 9th week in 100 % CD containing vermibin, whereas maximum urease activity was 1208 µg NH4 (-)N g(-1) h(-1) in 3rd week in 100 % CD containing vermibin. The enzyme activity and microbial counts were lesser in ETPS containing vermibins than control (100 % CD). The growth and fecundity of the worms in different vermibins were also investigated. The results showed that initially biomass and fecundity of the worms increased but decreased at the later stages due to non-availability of the palatable feed. This showed that quality and palatability of food directly affect biological parameters of the system.

Involvement of the iron regulatory protein from Eisenia andrei earthworms in the regulation of cellular iron homeostasis.

Iron homeostasis in cells is regulated by iron regulatory proteins (IRPs) that exist in different organisms. IRPs are cytosolic proteins that bind to iron-responsive elements (IREs) of the 5'- or 3'-untranslated regions (UTR) of mRNAs that encode many proteins involved in iron metabolism. In this study, we have cloned and described a new regulatory protein belonging to the family of IRPs from the earthworm Eisenia andrei (EaIRP). The earthworm IRE site in 5'-UTR of ferritin mRNA most likely folds into a secondary structure that differs from the conventional IRE structures of ferritin due to the absence of a typically unpaired cytosine that participates in protein binding. Prepared recombinant EaIRP and proteins from mammalian liver extracts are able to bind both mammalian and Eisenia IRE structures of ferritin mRNA, although the affinity of the rEaIRP/Eisenia IRE structure is rather low. This result suggests the possible contribution of a conventional IRE structure. When IRP is supplemented with a Fe-S cluster, it can function as a cytosolic aconitase. Cellular cytosolic and mitochondrial fractions, as well as recombinant EaIRP, exhibit aconitase activity that can be abolished by the action of oxygen radicals. The highest expression of EaIRP was detected in parts of the digestive tract. We can assume that earthworms may possess an IRE/IRP regulatory network as a potential mechanism for maintaining cellular iron homeostasis, although the aconitase function of EaIRP is most likely more relevant.

Bioremediation of distillery sludge into soil-enriching material through vermicomposting with the help of Eisenia fetida.

The aim of the present study was bioremediation of distillery sludge into a soil-enriching material. It was mixed with a complementary waste, cattle dung, and subjected to vermicomposting with (V) and without (T, control) Eisenia fetida in the ratio of 0:100 % (V1, T1), 10:90 (V2, T2), 25:75 (V3, T3), 50:50 (V4, T4), 75:25 (V5, T5) and 100:0 % (V6, T6), respectively. Survival rate, growth rate, onset of maturity, cocoon production and population build-up increased with increasing ratio of cattle dung. Maximum mortality of earthworm was observed in V6 mixture. On the basis of response surface design, the concentration of sludge giving highest number of worms, cocoons and hatchlings came out to be 21.11, 24.51 and 17.19 %, respectively. Nitrogen, phosphorus, sodium and pH increased during vermicomposting but decreased in the products without earthworm and there was increase in the contents of transition metals in the products of both the techniques. However, organic carbon, electrical conductivity and potassium showed an opposite trend.

Impact of fly ash and phosphatic rock on metal stabilization and bioavailability during sewage sludge vermicomposting.

Sewage sludge (SS) was mixed with different proportions of fly ash (FA) and phosphoric rock (PR), as passivators, and earthworms, Eisenia fetida, were introduced to allow vermicomposting. The earthworm growth rates, reproduction rates, and metal (except Zn and Cd) concentrations were significantly higher in the vermireactors containing FA and PR than in the treatments without passivators. The total organic carbon (TOC) and total metal concentrations in the mixtures decreased, and the mixtures were brought to approximately pH 7 during vermicomposting. There were significant differences in the decreases in the metal bioavailability factors (BFs) between the passivator and control treatments, and adding 20% FA (for Cu and Zn) or 20% PR (for Pb, Cd, and As) to the vermicompost were the most effective treatments for mitigating metal toxicity. The BF appeared to be dependent on TOC in the all treatments, but was not closely dependent on pH in the different vermibeds.

Non-hazardous anticancerous and antibacterial colloidal 'green' silver nanoparticles.

Poly(ethylene glycol) stabilized colloidal silver nanoparticles were prepared using the reductive potency of the aqueous extract of Thuja occidentalis leaves under ambient conditions. The nanoparticles were well dispersed within a narrow size spectrum (7-14 nm) and displayed characteristic surface plasmon resonance peak at around 420 nm and Bragg's reflection planes of fcc structure. MTT assay revealed the dose-dependent cytocompatibility and toxicity of the nanoparticles with the L929 normal cell line. On the other hand, the antiproliferative action of the nanoparticles was evaluated on HeLa cell (cancerous cells) line. Fluorescence and phase contrast microscopic imaging indicated the appearance of multinucleate stages with aggregation and nuclear membrane disruption of the HeLa cells post treatment with the nanoparticles. The interaction at the prokaryotic level was also assessed via differential antibacterial efficacy against Staphylococcus aureus (MTCC 3160) and Escherichia coli (MTCC 40). Under these perspectives, it is also necessary to observe the environmental impact of the prepared silver nanoparticles. Hence, the dose dependent toxicity of silver nanoparticles was evaluated upon the earthworm species Eisenia fetida. Neither the survival nor the reproduction was affected by the addition of silver nanoparticles up to 1000 ppm. Thus these 'green' silver nanoparticles have promising potential as future materials.

Vermicomposting of herbal pharmaceutical industry waste: earthworm growth, plant-available nutrient and microbial quality of end materials.

Efforts were made to decompose herbal pharmaceutical industrial waste (HPIW) spiked with cow dung (CD) using Eisenia fetida. A total of five vermibeds: T(1) - HPIW (0%+CD 100%, control), T(2) - HPIW (25%), T(3) - HPIW (50%), T(4) - HPIW (75%) and T(5) - HPIW (100%) were used for vermicomposting. The changes in biology and chemistry of vermibeds were measured after ten days interval. E. fetida showed high growth and cocoon production rate in all vermibeds. The vermicomposted material contained great population of fungi 6.0-40.6 (CFU × 10(5)g(-1)), bacteria 220-1276.0 (CFU × 10(8)g(-1)) and actinomycetes 410.0-2962.0 (CFU × 10(5)g(-1)) than initial material. Vermicomposted material was rich in plant-available forms of nutrients (N-NO(3)(-),PO(4)(3-),available K and SO(4)(-2)). Results suggested that noxious industrial waste can be converted into valuable product for sustainable soil fertility programme.

Simultaneous sludge reduction and nutrient removal (SSRNR) with interaction between Tubificidae and microorganisms: a full-scale study.

A symbiotic ecosystem between Tubificidae and microorganisms was established at a full-scale wastewater treatment plant (WWTP). In this ecosystem Tubificidae were inoculated, and then adhered to the outer layers of carrier materials in an oxidation tank. During the long-term treatment of sewage volumes of 20,000 m(3)d(-1), the excess sludge production rate was reduced from 0.21 to 0.051 kg m(-3) and sludge settleability was significantly improved. When the influent concentrations of COD, NH(4)(+)-N, PO(4)(-)-P, and SS were in the ranges of 130.0-459.0 mg L(-1), 14.2-27.5 mg L(-1), 1.6-7.0 mg L(-1), and 60.0-466.0 mg L(-1), respectively, the COD and SS removal efficiency was increased by 8.7% and 13.6% within the symbiotic system compared to the control without Tubificidae. In addition, NH(4)(+)-N and phosphorus removal efficiency can also be improved. The results showed that both sludge reduction and nutrient removal were enhanced simultaneously significantly within the system utilizing the symbiotic interactions of Tubificidae and microorganisms.

The potential reuse of soybean husk as feedstock of Eudrilus eugeniae in vermicomposting.

BACKGROUND: Soybean (Glycine max L.) is one the most commonly consumed legumes worldwide, with 200 million metric tons produced per year. However, the inedible soy husk would usually be removed during the process and the continuous generation of soybean husk may represent a major disposal problem for soybean processing industries. Thus, the main aim of the present study was to investigate the possibility to convert soybean husk (S) amended with market-rejected papaya (P) into vermicompost using Eudrilus eugeniae. RESULTS: Soybean husk was mixed with market-rejected papaya in different ratios to produce five different treatments (1P:1S, 2P:1S, 1P:2S, S only and P only) for laboratory screening of solid wastes. Generally, the application of E. eugeniae permitted a significant increase in Ca (42.3-91.6%), K (93.8-235%), Mg (25.4-84.6%) and P (37.1-129%) but a decrease in the C:N ratio (21.4-52.8%) after 9 weeks of vermicomposting. Among all the treatments investigated, soybean husk which was mixed with an equal amount of market-rejected papaya (1P:1S) showed superior quality vermicompost. Waste mixtures also encouraged the growth of E. eugeniae except the treatments with soybean husk alone as well as soybean husk which was mixed with market-rejected papaya in 1P:2S ratio. CONCLUSION: Vermicomposting could be used as an efficient technology to convert soybean husk into nutrient-rich organic fertiliser if it was mixed with market-rejected papaya in appropriate ratio.

Evaluation of three composting systems for the management of spent coffee grounds.

This study was conducted to evaluate the optimum composting approach for the management of spent coffee grounds from the restaurant and ready-to-serve coffee industry. Three composting systems were assessed, including in-vessel composting, vermicomposting bins, and aerated static pile bin composting, over study periods ranging from 47 to 98 days. Total carbon content was reduced by 5-7% in the spent coffee ground treatments across the three composting systems. Nitrogen and other mineral nutrient contents were conserved or enhanced from the initial to the final composts in all the composting systems assessed. Earthworm growth and survival (15-80%) was reduced in all the treatments but mortality rates were lower in coffee treatments with cardboard additions. A decline in earthworm mortality with cardboard additions was the result of reduced exposure to organic compounds and chemicals released through the decomposition of spent coffee grounds.

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.