Behaviour Of Earthworms Research Articles

Short- and long-term impacts of anaerobic digestate spreading on earthworms in cropped soils

Anaerobic digestion is increasingly used in Europe to treat organic substrates and produce biogas as a renewable energy source. The residual matter (digestate) is used in agriculture as an organic fertilizer. The study aims at assessing the impact of digestate application in the field on earthworms from the short term (few hours) to the long term (two years), and at investigating under laboratory conditions the role of ammonia and earthworm behavior on digestate toxicity in the short term. First, we studied earthworm communities in fields fertilized with digestates, cattle effluents, or chemical fertilizers for two years. Earthworm abundance was assessed before and after the fertilization event of the third year. Earthworm mortality at the soil surface was also assessed immediately after fertilization. Next, the toxicity of digestate or ammonia solutions on Aporrectodea caliginosa and Lumbricus terrestris was measured in microcosms (110 cm3) to better understand the short-term toxicity (two weeks). Finally, we spread digestate (40–80 t ha−1) on soil columns (5300 cm3) and used X-ray tomography after two weeks to assess the burrowing behavior of earthworms in the cores. Earthworm abundance was 150% higher in the fields treated for two years with digestates or cattle effluents compared to the field treated with chemical fertilizers. 0.5 to 2% of adult earthworms died at the soil surface a few hours after liquid digestate and cattle slurry spreading (18 to 24 t ha−1). The digestate (10% to 20% (fresh digestate/dry soil)) and ammonia were also lethal to earthworms in the microcosms within two weeks. In contrast, no mortality occurred inside soil columns two weeks after digestate spreading; A. caliginosa avoided the soil surface with high digestate inputs. This case study highlighted the potential short-term toxicity of digestate (a few hours), which evolved towards a neutral to positive impact in the field in the longer term (from two weeks to two years). Further research is needed to understand the impact of diverse solid and liquid digestates on soil macrofauna in different soils.

Histological study on the effect of sodium bentonite nanoparticles on the earthworm, Allolobophora caliginosa (Lumbricidae: Annelida)

Bentonites are used on a large scale, by farmers and the feed industry, as a feed supplements to adsorb mycotoxins. The presence of mycotoxins in feed affects its quality, animal safety, and human health. Food additives must be used carefully as they may contaminate food and became harmful to the environment and food consumers. The present work studied the effect of two different concentrations of sodium bentonite on the body wall, gut, and coelomic cells of earthworms. The results revealed a change in the behavior of earthworms and the occurrence of a great alteration in the tested samples but no mortality was recorded. Coelomic cells aggregated, vacuoles and pseudopodia were formed.

Current Research Trends on the Effects of Microplastics in Soil Environment Using Earthworms: Mini-Review

Objectives : The increase in the amount of plastics used has brought convenience to human life, but has also caused various environmental problems such as problems related to microplastics. Microplastics are defined as small plastic particles with a size of under 5 mm. As of now, research on microplastics has been largely focusing on the marine environment, and studies on microplastics in soil environment are on the rise in very recent years. Thus, the environmental impact of microplastics on the soil environment are insufficient than those on the marine environment, and there is a need for more studies. Therefore, this mini-review attempts to examine the studies on microplastics in soil environment focusing on the studies using earthworms as a test species.Methods : Published studies on microplastics in soil environment were identified by searching Web of Science. The focus of this mini-review was to gather the published articles reporting the effect of microplastics on earthworms in the absence and presence of other chemical contaminants.Results and Discussions : The number of studies evaluating the effect of microplastics in soil are significantly increasing, and earthworms are being mostly used as the test species. Experimental conditions were divided into two types: Single exposure to microplastics and combined exposure to microplastics and other chemicals. In most studies, changes in the biomarkers which are related to oxidation stress of earthworms were confirmed, and direct damage to intestinal tissues of earthworms was also observed. However, in the case of the changes of the biomarkers of earthworms, their patterns were different depending on the experimental conditions (i.e., concentrations of microplastics and chemicals, microplastics size and type, exposure period, etc.). Also, the avoidance behavior of earthworms and the internal accumulation of microplastics and co-existing chemicals were not consistent among the studies. Thus, the effects of microplastics on earthworms are not clear yet, although they seem to have some adverse effects.Conclusions : This study showed that the interest in the impact of microplastics in the soil environment has been increased recently, and earthworms are being used mostly in the studies. Yet, there is no standardized method to study the effect of microplastics on earthworms. Studies so far confirmed that microplastics induced changes in and outside of earthworm bodies. However, due to various experimental conditions, it seemed that consistent results could not be confirmed in those researches. For that reason, this study suggests that standardized research methods for microplastics researches using earthworms as a test species need to be established and need to be applied to further studies.

Effect thresholds for the earthworm Eisenia fetida: Toxicity comparison between conventional and biodegradable microplastics

Biodegradable plastics have been developed to eliminate the progressive accumulation and ever-growing threat posed by conventional fossil fuel-derived plastics. The impact of these bioplastics, particularly in an agricultural context (e.g. biopolymer mulch films), however, remains poorly understood. In this study, we compared the biotoxicity of biodegradable (polylactic acid, PLA; polypropylene carbonate, PPC) and non-degradable (polyethylene, PE) microplastics using a series of standardized bioassays using the earthworm Eisenia fetida. The responses studied included: avoidance behavior, mortality, biomass, and reproduction responses. We incubated earthworms in artificial soils amended with different concentrations of microplastic (0, 0.125, 1.25, 12.5, 125, 250, and 500 g kg−1) under laboratory conditions. This wide range allowed linear regression modeling and estimation of microplastic effect thresholds. Our results showed that microplastic concentration rather than plastic type was more important in regulating earthworm responses to soil contamination. The critical threshold for microplastic contamination was 40 g kg−1, after which earthworms exhibit microplastic avoidance behavior. A significant reduction (EC10) in number of cocoons and juvenile earthworms occurred at a concentration of 53 g kg−1 and 97 g kg−1, respectively; while no significant effect was found for survival of earthworm until levels of 500 g kg−1. Overall, the two biodegradable materials (PLA and PPC), appeared to be no more biofriendly than PE. Based on reported levels of plastic contamination in soil of up to 67 g kg−1, we conclude that microplastics are now starting to pose a threat to earthworm population. To better evaluate the risk posed by biodegradable and nondegradable plastics, further mechanistic studies on how microplastics affect earthworm behavior and the potential long-term impacts of this on soil functioning are required.

Decreased burrowing activity of endogeic earthworms and effects on water infiltration in response to an increase in soil bulk density

Endogeic earthworms live and burrow in the soil to find their food. They burrow by pushing the soil aside or ingesting it and are thus sensitive to soil compaction. However there is a scarcity of data regarding the effects of soil bulk density on the burrowing behavior and activity of endogeic earthworms. We carried out laboratory experiments using repacked soil cores with various levels of bulk density (from 1.18 to 1.38 g cm−3) in which individuals of Aporrectodea caliginosa or Allolobophora chlorotica were incubated for six weeks. The burrow systems inside the soil cores and the compaction around the burrows were then analyzed using X-ray tomography. Soil water infiltration measurements were also carried out. The increase in bulk density had a negative impact on all burrow system characteristics (length, volume, diameter, continuity, number of burrows). When bulk density increased from 1.18 to 1.38 g cm−3, volume, diameter, continuity and the number of burrows decreased on average by 77 %, 21 %, 81 % and 58 %, respectively. The increase in density due to compaction around the burrows was similar whatever the species and the bulk density. Increasing soil bulk density from 1.18 to 1.38 g cm-3 also greatly decreased water infiltration (−89 % for both species) and increased breakthrough time (10 and 25-fold for A. chlorotica and A. caliginosa respectively). However, compared to a control without earthworms, water infiltration in cores incubated with endogeic species was only increased significantly at 1.18 and 1.23 g cm−3. This illustrates that burrows made by endogeic earthworms moderately increase water infiltration and only when the soil bulk density is low. Data provided in this study could be used to refine simulation models of earthworm burrowing behavior where burrowing is assumed to be mainly governed by soil water content, temperature and soil bulk density.

Ecotoxicological responses of Eisenia andrei exposed in field-contaminated soils by sanitary sewage

The disposal of untreated sanitary sewage in the soil has several consequences for human health and leads to environmental risks; thus, it is necessary investigating, monitoring and remediating the affected sites. The aims of the current study are to evaluate ecotoxicological effects on Eisenia andrei earthworms exposed to soil subjected to sources of sanitary sewage discharge and to investigate whether prevention values established by the Brazilian legislation for soil quality, associated with the incidence of chemical substances in it, are satisfactory enough to assure the necessary quality for different organisms. Earthworms’ behavior, reproduction, acetylcholinesterase activity, catalase, superoxide dismutase and malondialdehyde levels were evaluated. The reproduction and behavior of earthworms exposed to sanitary sewage were adversely affected. Increased superoxide dismutase and catalase activity acted as antioxidant defense mechanism. Significantly increased lipid peroxidation levels and acetylcholinesterase activity inhibition have indicated lipid peroxidation in cell membrane and neurotransmission changes, respectively. Results have confirmed that sanitary sewage induced oxidative stress in earthworms. In addition, based on biochemical data analysis, the integrated biomarker response (IBR) has evidenced different toxicity levels in earthworms between the investigated points. Finally, results have indicated that effluents released into the soil, without proper treatment, lead to contaminant accumulation due to soil saturation and it can hinder different processes and biological development taking place in the soil. In addition, the current study has shown that physical-chemical analyses alone are not enough to assess soil quality, since it is also requires adopting an ecotoxicological approach. Brazilian legislation focused on soil quality must be revised and new guiding values must be proposed.

Ecotoxicity of imidacloprid to soil invertebrates in two tropical soils with contrasting texture.

Imidacloprid is one of the most commercialized insecticides in agriculture in the world, with a broad spectrum of action. However, little is known about the effects of commercial formulations containing this active ingredient (a.i.) on non-target organisms in tropical soils. Our objective was to assess the toxicity based on the predicted environmental concentration (PEC) of imidacloprid, in the avoidance behaviour of earthworms and collembolans as well as in the reproduction of collembolans, in two representative soils of the Brazilian Cerrado with contrasting texture (clayey Oxisol and sandy Entisol). Ecotoxicity tests were carried out according to ISO protocols to assess the avoidance behaviour of earthworms (Eisenia andrei) and avoidance and reproduction of collembolans (Folsomia candida). In the earthworm's avoidance test, more than 80% of the individuals were found in the control, in all tested concentrations, indicating a possible habitat function loss in both soils. The avoidance behaviour of collembolans was observed in both soils, being more expressive (up to 75% of escape) in Oxisol. In Entisol, only the two highest concentrations were avoided (up to 63%). There was a negative effect on the reproduction of collembolans in both soils, with a higher EC50 value (0.255 mg kg-1) in Oxisol than in Entisol (0.177 mg kg-1), demonstrating higher toxicity in the sandy soil. These differences were attributed to the contrasting texture of the studied soils, probably due to lower retention of the a.i. in the sandy soil, causing an increased bioavailability. This study demonstrated that imidacloprid can be highly toxic to soil invertebrates, even in soil concentrations lower than those expected from recommended dose, causing an impact on the edaphic organisms and, consequently, compromising its functions in the soil ecosystem.

Depth distribution of soil organic matter and burrowing activity of earthworms—mesocosm study using X-ray tomography and luminophores

Earthworms feed on organic matter present at the soil surface or within the soil. Thus, its distribution in the soil profile is likely to greatly influence earthworm behavior and, in turn, their burrow system. To test this idea, two anecic and two endogeic earthworm species were introduced into repacked soil cores (depth = 30 cm) upper half filled with a top soil containing 4% organic matter (0–15 cm) and lower half filled with a deep soil at 2% organic matter (15–30 cm). Earthworm behavior was studied using X-ray tomography combined with luminophores (colored particulate tracers of 63–125 μm size) placed at 0, 3, and 12 cm depth, a method widely used in sediment ecology. We observed that anecic and endogeic earthworms had contrasting reactions to the conditions with only endogeic species burrowing more intensively in the upper part. From a quantitative point of view, only a few percent of luminophores were displaced. However, luminophore displacements also provided qualitative information to complement the tomography: (i) endogeic species and especially Aporrectodea caliginosa bioturbated the most soil close to the surface (3 cm depth) and (ii) the two anecic species influenced the luminophore distribution differentially with Lumbricus terrestris displacing significantly more luminophores, whatever their initial depth, than Aporrectodea nocturna due to intense surface cast activity. Beyond methodological developments, our study found that endogeic earthworms burrow more in zones with higher organic matter contents and this explains why they are mainly found close to the soil surface in non-tilled soils.

Earthworm activity and its coupling to soil hydrology: A deterministic analysis

Considering in situ observations, chaos theory was taken as a basis to study the activity of anecic earthworms based on cast production from September 2016 to January 2018 in the Dong Cao watershed (Vietnam). To study this activity, the global modeling technique was used to obtain deterministic models of ordinary differential equations directly from observational time series. The obtained models show that the behavior of earthworms is chaotic; it is coupled to the dynamics of soil water content in a complex (integrative) way and can be interpreted as an habituation/sensitization process. However, this coupling is insufficient to explain the desynchronization of the cast production variations observed at different study sites. The retroaction of earthworm activity on soil is required to explain the spatiotemporal discrepancies.

Ecotoxicological assessment of silicate rock fertilizers using soil invertebrates

Abstract: The objective of this work was to evaluate the possible adverse effects of rock dusts on the avoidance behavior of earthworms and collembolans, as well as on the survival and reproduction of enchytraeids, in ecotoxicological tests using artificial and natural soils. The tests were performed following international guidelines (ISO 17512-2/2011, ISO 11267/1999, and NBR ISO 16387/2012), with some adaptations, in order to evaluate the effect of dusts from two silicate rocks, biotite schist and phonolite, at the concentrations of 0, 0.1, 1.0, 10, and 100%, in artificial and natural (Arenosol and Ferralsol) soils. The avoidance behavior of the earthworm Eisenia andrei and the collembolan Folsomia candida was evaluated, as well as the survival and reproduction of the enchytraeid Enchytraeus sp. Biotite schist and phonolite affected negatively earthworms when applied to the soil at concentrations equal to or higher than 1.0%. The observed effect was associated to the greater presence of salts, especially in the phonolite, and to changes in soil texture when higher concentrations were used. Negative effects on collembolans and enchytraeids were verified at concentrations of 10% or using pure rock dusts. Agronomic rates smaller than 1.0% (approximately 20 Mg ha-1) rock dusts do not cause avoidance behavior or reduction in the survival and reproduction of invertebrates in the soil.

Effect of chlorpyrifos and carbofuran on morphology, behavior and acetylcholinesterase activity of earthworm (Eisenia fetida)

Organophosphates (OP) and carbamates when applied in agricultural fields are the potential environmental polluters and toxicants for soil flora and fauna. Upon exposure to such chemical pesticides, morphology, behaviour and acetylcholinesterase (AChE) activity of earthworms (Eisenia fetida) is altered. The present study dealt with the comparative analysis of toxic effects induced by chlorpyrifos (OP) and carbofuran (carbamate) exposure on morphology, behavior and AChE activity using standard filter paper contact toxicity method. The LC50 of chlorpyrifos and carbofuran for 24 hr was 0.25% and 5.13%, respectively, marking chlorpyrifos as more toxic pesticide than carbofuran. The treated worms exhibited abnormal morphological symptoms such as excessive mucus secretion, depigmentation, deformity, loss of metameric segments and damaged clitellum. Variation in behaviour such as reduced activity, sluggish movements and flattened posture marked the stress induced due to pesticide toxicity even at lower doses. Restlessness along with jerky movements was observed on exposure to higher concentrations of chlorpyrifos, whereas lower concentrations resulted in avoidance behavior toward pesticide coated glass vials. Thereafter, worms were exposed to different concentrations of chlorpyrifos (0.13%, 0.25% and 0.38%) and carbofuran (2.57%, 5.13% and 7.70%) for 24h and 48h to assess sub-acute and acute toxicity effects of these pesticides on AChE. The enzyme activity after 48h was 94.14%, 60.21% and 40.44% for the worms exposed to 0.125%, 0.25% and 0.375% chlorpyrifos, respectively. The enzyme activity after 48h was 98.17%, 93.92% and 79.25% for the worms exposed to 2.57%, 5.13% and 7.70% carbofuran, respectively. Time and dose dependent significant (p<0.05) decrease in the levels of AChE in pre-clitellar region of earthworms was observed as compared to control when worms were exposed to chlorpyrifos and carbofuran. Alteration in behavioural response of the earthworms may be attributed to the decline in AChE activity of pre-clitellar region. However, chlorpyrifos was more potent inhibitor of AChE activity in Eisenia fetida as compared to that of carbofuran. Such alterations indicate the potential health risk of these pesticides’ exposure at high concentrations to E. fetida.

Physiological Responses of Earthworm Under Acid Rain Stress.

Acid rain has become one of the major global environmental problems, and some researches reported that acid rain may have a certain inhibition on soil biodiversity. Besides this, it is well known that earthworm (Eisenia fetida) plays an important role in the functioning of soil ecosystems. For this point, we conducted a series of experiments to investigate whether acid rain would take effects on earthworms. In the present study, the earthworms were incubated on filter paper and in soil under acid rain stress. The mortality and behavior of earthworms were recorded, and epidermal damage and the activity of the CYP3A4 enzyme were measured for the tested earthworms. Our experimental results showed that the earthworms could not survive in the acid rain stress of pH below 2.5, and acid rain with weak acidity (i.e., 4.0 ≤ pH ≤ 5.5) promoted the activity of the CYP3A4 enzyme in the earthworms, while acid rain with strong acidity (i.e., 3.0 ≤ pH ≤ 3.5) inhibited it. Moreover, the degree of damage in sensitive parts of the earthworms increased with the decrease of pH value. This study suggests that acid rain can cause discomfort response and the direct epidermal damage of earthworms, and even kill them.

Using X-ray microtomography to characterize the burrowing behaviour of earthworms in heterogeneously polluted soils

Large amounts of chemicals are released into soil environments due to human activities, thereby harming organisms and their habitats. Earthworms are important bioindicators for chemical risk assessments of soil ecosystems, but the question remains of how they respond to different patterns of pollution. Laboratory experiments in soil mesocosms were carried out to observe changes in the burrowing behaviour of two earthworm species (Aporrectodea caliginosa and Lumbricus terrestris) with regard to different spatial distributions of benzo(a)pyrene, cadmium and copper, using X-ray computed microtomography (μCT). This is, to our knowledge, the first contribution of μCT to study earthworm behaviour in heterogeneously multi-polluted soils. Columns (14 × 30 cm) were filled with a silt loam soil and packed to represent two scenarios of heterogeneous pollution: a) ‘binary’, containing polluted soil in the low half part and unpolluted soil on top; and b) ‘patches’, containing small portions of polluted soil randomly placed into an unpolluted matrix. Earthworms were introduced individually in the columns and left in climatic chambers for 7 weeks. μCT images revealed that earthworms avoided the polluted zones differently (e.g. A. caliginosa>L. terrestris, ‘binary’ configuration). Individuals of L. terrestris that entered polluted areas showed a lower burrowing activity (volume and length of the burrow system) than those that did not enter. The presence of ‘patches’ of polluted soil did not cause a reduction of the burrowing activity such as that found in the ‘binary’ columns, although earthworms showed avoidance. Our results indicate that pollutants had a greater effect on earthworms’ burrowing behaviour in the presence of larger portions of contaminated soil compared with small random ones. The more evident avoidance behaviour of A. caliginosa pinpoints its usefulness in toxicity bioassays based on avoidance patterns, but questions its use in assays based on bioaccumulation of pollutants in tissues, for which L. terrestris may perform better.

Observing earthworm behavior using deep learning

Earthworm behavior represents a valuable endpoint in ecological and ecotoxicological studies, providing insight into individual- and population-level responses of organisms to ecological factors and xenobiotics. Difficulties in observing activities of soil-dwelling organisms come from the soil matrix surrounding the animals. We present an automated system for continuous monitoring of earthworm behavior directly in the soil. Performances of the developed system are tested in an avoidance test set-up. The developed system includes a 2D terrarium serving as an experimental compartment and a deep convolutional neural network model for continuous monitoring of earthworm behavior. Quantification of activity of organisms is based on the predictions of the neural network model made from image sequences captured during the exposure. Performance of the system was validated by comparison with results of the standard avoidance test, using H3BO3, a standard pollutant. The presented system represents a simple, cost-effective, fast, accurate and objective tool for continuous earthworm behavior monitoring in ecological and ecotoxicological studies. Source code and training data of this system are made freely available on GitHub.

A novel method for real-time monitoring of soil ecological toxicity – Detection of earthworm motion using a vibration sensor

The aim of this study was to develop a new method, using a vibration sensor, to address the drawbacks of preexisting methods for monitoring soil ecological toxicity. A novel method was designed by inspiration from seismometers, which record signals originating from the ground motion caused by earthquake events. Similarly, the newly developed method using a vibration sensor detects the signals generated by earthworm activity, which reflects the soil ecological toxicity. To establish the new method, a stepwise approach was adopted: (1) the effects of operational conditions on the overall performance of the system were evaluated, and (2) the feasibility of the method was tested by an application study. A number of crucial factors influencing the overall performance of the method were evaluated. These were categorized based on three features: soil, tested organism, and instrumentation. The soil properties evaluated included soil type (artificial and natural), moisture content, and bulk density. In terms of the organism, the effect of the number of earthworms was investigated. Finally, with regard to instrumentation, appropriate soil chamber specifications and monitoring duration were identified. The most effective conditions for each factor were determined based on a comparative evaluation of changes in the activity levels and body weights of the earthworms. After the first step of the study, an application study was carried out to demonstrate the feasibility of the proposed method. Zinc (Zn)-contaminated soils were tested under the most efficient operational conditions identified in the preceding study. The results of the study confirm that the method is applicable to natural soils, and the best performance was achieved under soil conditions of 50–60% maximum water holding capacity and 0.95 g/cm3 bulk density. Furthermore, the optimal number of earthworms was found to be five, which corresponds 19.84 g soil per earthworm. With respect to the instrumental conditions, the most efficient specification was a cylindrical soil chamber with a diameter of 94 mm and height of 54 mm. Additionally, the most relevant monitoring duration was found to be 7 days. The results indicate that the method can shorten the testing period, reduce the soil amount and earthworm number required, and facilitate the real-time monitoring of mortality. Based on the results of the application study, we validated the proposed vibration sensor-based method for characterizing earthworm behavior in terms of its feasibility for monitoring the ecological toxicity of soil. The results indicate that dermal contact and feeding activity of earthworms decreased significantly with increasing Zn concentrations in the soil. The EC50 value of Zn calculated based on the earthworm behavior was 340.97 mg/kg. Based on the results, it is concluded that the proposed method cannot only overcome the shortcomings of traditional test methods using earthworms, but also enable real-time ecotoxicity in soil environments.

Ecotoxicological risk assessment of contaminated soil from a complex of ceramic industries using earthworm Eisenia fetida

ABSTRACTThe aim of this study was to determine ecotoxicological parameters for biomonitoring of environmental risk of native soils from a ceramic industrial area that had been contaminated with cadmium (Cd) and chromium (Cr) by using the earthworm, Eisenia fetida. Initially, lab tests were conducted to compare earthworm (Eisenia fetida) growth, survival, morphology, behavior, and reproduction rates following exposure to six concentrations of contaminated soil at 0%, 6.25%, 12.5%, 25%, 50%, or 100% mixed in artificial soil and cow dung following a 28-d incubation period. The second experiment consisted of utilizing Eisenia fetida in a predetermined lowest observed effect concentration to measure heavy metals bioaccumulation from superficial soil collected from a ceramic industrial area following a 56-d exposure. Data demonstrated that in the lab earthworms maintained at 6.25% of contaminated soil, exhibited significant increase in mean weight, bioaccumulation of Cd and Cr associated with a significant decrease in the amount of Cd and Cr in the soil. At field testing, similar results that were observed as in the lab as evidenced by rise in mean weight, higher levels of Cd and Cr in the earthworm tissue accompanied by significant fall in soil levels of Cd and Cr. In conclusion, at tested relevant environmental concentrations, the use of Eisenia fetida for assessing ecotoxicological risk arising from contaminated soil due to ceramic industrial pollutant emissions was found to be an effective tool for biomonitoring program.

Sub-lethal effects of six neonicotinoids on avoidance behavior and reproduction of earthworms (Eisenia fetida)

Avoidance behavior of earthworms (Eisenia fetida) against six neonicotinoids (NEOs) (acetamiprid, dinotefuram, clothianidin, thiacloprid, nitenpyram, imidacloprid) was studied following the protocol of ISO. The results showed obvious avoidance behavior of E. fetida against the tested insecticides, and the medium effective concentration for avoidance behavior (EC50) of the six pesticides was 0.14, 0.55, 0.91, 7.87, 1.32 and 0.77 mg/kg, respectively. Compared to the acute toxicity, avoidance behavior was more sensitive as an indicator of soil contamination with NEOs. Chronic toxicity of above six NEOs to E. fetida was also evaluated; cocoon production, hatchability, cocoon weight and adult weight were all affected in the test. Cocoon production and hatchability were more sensitive than cocoon weight and adult weight. The reproduction of earthworms were significantly reduced with a 56 d half-maximal effective hatchability concentration (EC50) of 0.37, 0.74, 1.30, 3.57, 1.20 and 0.70 mg/kg (acetamiprid, dinotefuram, clothianidin, thiacloprid, nitenpyram, imidacloprid), respectively. Most of the tested NEOs were highly toxic to E. fetida. Avoidance behavior and reproduction damage of E. fetida was observed at very low concentrations. The existing levels of pollution with NEOs in soil frequently exceed the lowest observed adverse effect concentrations, which are likely to have negative biological and ecological impacts on earthworms.

Water treatment residuals as soil amendments: Examining element extractability, soil porewater concentrations and effects on earthworm behaviour and survival

Drinking water treatment residuals (WTRs), the by-product of water clarification processes, are routinely disposed of via landfill however there is a growing body of research that demonstrates the material has great potential for beneficial use in environmental applications. Application to agricultural land is one option showing great promise (i.e. a low cost disposal route that provides organic matter input to soils and other potential benefits), however questions remain as to the impact such applications may have on earthworm survival and behaviour and also on the potential effects it may have on soil porewater chemistry. This study examined the leachability of elements within two types of WTRs (one Al- and one Fe- based) from England via 0.001 M CaCl2 solution, at varying pH, and via the Community Bureau of Reference (BCR) sequential extraction scheme. Earthworm avoidance, survival, growth, reproduction and element concentrations were examined in WTR-amended sandy soils (0%, 5%, 10%, 20% w/w), while soil porewaters were also recovered from experimental units and examined for element concentrations. The results revealed leachable element concentrations to be very low in both types of WTRs tested and so element leaching from these WTRs would be unlikely to pose any threat to ecosystems under typical agricultural soil conditions. However, when the pH was lowered to 4.4 there was a substantial release of Al from the Al-WTRs (382 mg/kg). Soil porewater element concentrations were influenced to some degree by WTR addition, warranting further examination in terms of any potential implications for nutrient supply or limitation. Earthworm avoidance of WTR-amended soil was only observed for Al-WTRs and only at the maximum applied rate (20% w/w), while survival of earthworms was not affected by either WTR type at any application rate. Earthworm growth and reproduction (cocoon production) were not affected at a statistically significant level but this needs further examination over a longer period of exposure. Increased assimilation of Al and Fe into earthworm tissues was observed at some WTR application rates (maximum fresh weight concentrations of 42 mg/kg for Al and 167 mg/kg for Fe), but these were not at levels likely to pose environmental concerns.

Biaccumulation and tolerance of heavy metals on the tropical earthworm, Allobophora sp. after exposed to contaminated soil from oil mine waste

In this study, the impact of contaminated soil from oil mine waste on survival, behavior, tolerance, and bioaccumulation of heavy metals by the tropical earthworm, Allobophora sp. has been quantified. Earthworm was isolated from heavy metals-contaminated soil, cultured in laboratory condition, and exposed to contaminated soil from oil mine waste for a couple of months. The behavior and response of earthworms to contaminated soil was monitored for 28 days and evaluated by the response criteria was expressed in scale index (SI) referred to Langdon method. Resistance test of the earthworm (LC50) to heavy metals also conducted with variation soil concentrations of 100%, 50%, 25%, 12.5%, and 6.25%, and 0% (Control). Results showed that contaminated soil extremely affected to the earthworm live, especially length and their body weight. The Lethal Concentration 50% (LC50) of earthworm against contaminated soil was 19.05% (w/w). When exposed to contaminated soil, earthworm accumulated chromium, barium, and manganese at the concentration of 88; 92.2; and 280 mg/kg-DW, respectively. Based on these results, earthworm Allobophora sp. has potential to reduce heavy metals from contaminated soil in the field of bioremediation process.

Do recommended doses of glyphosate-based herbicides affect soil invertebrates? Field and laboratory screening tests to risk assessment

Despite glyphosate-based herbicides are widely used in agriculture, forestry and gardens, little is known about its effects on non-target organisms. The present work evaluated the ecotoxicity of four formulated products (Roundup® Original, Trop®, Zapp® Qi 620 and Crucial®) on soil invertebrates. Screening ecotoxicity tests were carried out with soil and oat straw collected in a field experiment, besides laboratory-spiked soils. Screening tests included avoidance behaviour of earthworms (Eisenia andrei), collembolans (Folsomia candida) and isopods (Porcellio dilatatus) in single and multispecies tests; reproduction of collembolans (F. candida), and bait lamina in field. Non-avoidance behaviour was observed in standard tests (earthworms) in soil, neither in multispecies tests (earthworm + isopods) using oat straw, while for collembolans it occurred for the product Zapp® Qi 620 even at the recommended dose. Reproduction of F. candida was not impaired even at high doses in laboratory-spiked soils. Feeding activity on bait lamina test was impaired in treatment corresponding to the red label product, Crucial®. Results showed the relevance of bait lamina test on screening the impact of herbicides in the field. The findings highlight the importance of considering different formulations for the same active ingredient in risk assessment of pesticides.