Skip to main content

Advertisement

SpringerLink
Log in

Sewage sludge toxicity assessment using earthworm Eisenia fetida: can biochemical and histopathological analysis provide fast and accurate insight?

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Sewage sludge (SS) is a complex organic by-product of wastewater treatment plants. Deposition of large amounts of SS can increase the risk of soil contamination. Therefore, there is an increasing need for fast and accurate assessment of SS toxic potential. Toxic effects of SS were tested on earthworm Eisenia fetida tissue, at the subcellular and biochemical level. Earthworms were exposed to depot sludge (DS) concentration ratio of 30 or 70 %, to undiluted and to 100 and 10 times diluted active sludge (AS). The exposure to DS lasted for 24/48 h (acute exposure), 96 h (semi-acute exposure) and 7/14/28 days (sub-chronic exposure) and 48 h for AS. Toxic effects were tested by the measurements of multixenobiotic resistance mechanism (MXR) activity and lipid peroxidation levels, as well as the observation of morphological alterations and behavioural changes. Biochemical markers confirmed the presence of MXR inhibitors in the tested AS and DS and highlighted the presence of SS-induced oxidative stress. The MXR inhibition and thiobarbituric acid reactive substance (TBARS) concentration in the whole earthworm’s body were higher after the exposition to lower concentration of the DS. Furthermore, histopathological changes revealed damage to earthworm body wall tissue layers as well as to the epithelial and chloragogen cells in the typhlosole region. These changes were proportional to SS concentration in tested soils and to exposure duration. Obtained results may contribute to the understanding of SS-induced toxic effects on terrestrial invertebrates exposed through soil contact and to identify defence mechanisms of earthworms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Achard M, Baudrimont M, Boudou A, Bourdineaud JP (2004) Induction of a multixenobiotic resistance protein (MXR) in the Asiatic clam Corbicula fluminea after heavy metals exposure. Aquat Toxicol 67(4):347–357

    Article  CAS  Google Scholar 

  • Amorim MJB, Novais S, Römbke J, Soares AMVM (2008) Enchytraeus albidus (Enchytraeidae): a test organism in a standardised avoidance test? Effects of different chemical substances. Environ Int 34:363–371

    Article  Google Scholar 

  • Antolin MC, Pascual I, Garcia C, Polo A, Sanchez-Diaz M (2005) Growth, yield and solute content of barley in soils treated with sewage sludge under semiarid Mediterranean conditions. Field Crops Res 94:224–237

    Article  Google Scholar 

  • Artuso N, Kennedy TF, Connery J, Grant J, Schmidt O (2011) Effects of biosolids at varying rates on earthworms (Eisenia fetida) and springtails (Folsomia candida). Appl Environ Soil Sci. doi: 10.1155/2011/519485

  • Aydoğan Kılıç G (2011) Histopathological and biochemical alterations of the earthworm (Lumbricus terrestris) as biomarker of soil pollution along Porsuk River Basin (Turkey). Chemosphere 83:1175–1180

    Article  Google Scholar 

  • Babić S, Dragun Z, Sauerborn Klobučar R, Bačić N, Fiket Ž, Barišić J, Krasnići N, Strunjak-Perović I, Topić Popović N, Čož-Rakovac R (2015) Different patterns of concentration variability of various metals and metalloids in the soluble tissue fractions of earthworm Eisenia fetida after exposure to active sludge. Sci Total Environ 526:127–135

    Article  Google Scholar 

  • Baynes RE, Hodgson E (2004) Absorption and distribution of toxicants. In: Hodgson E (ed) A textbook of modern toxicology, 3rd edn. Wiley & Sons, New York, pp 77–110

    Google Scholar 

  • Bošnjak I, Bielen A, Babić S, Šver L, Popović NT, Strunjak-Perović I, Čož-Rakovac R, Sauerborn Klobučar R (2014) First evidence of the P-glycoprotein gene expression (P-gp/Abcb1) and MXR activity modulation in earthworm Eisenia fetida, Savigny. Arh Hig Rada Toksikol 65(1):67–74

    Google Scholar 

  • Braguglia CM, Coors A, Gallipoli A, Gianico A, Guillon E, Kunkel U, Mascolo G, Richter E, Ternes TA, Tomei MC, Mininni G (2014) Quality assessment of digested sludges produced by advanced stabilization processes. Environ Sci Pollut Res. doi: 10.1007/s11356-014-3090-6

  • Caminada D, Žaja R, Smital T, Fent K (2008) Human pharmaceuticals modulate P-gp1 (ABCB1) transport activity in the fish cell line PLHC-1. Aquat Toxicol 90:214–222

    Article  CAS  Google Scholar 

  • Carpené E, Andreani G, Monari M, Castellani G, Isani G (2006) Distribution of Cd, Zn, Cu and Fe among selected tissues of the earthworm (Allolobophora caliginosa) and Eurasian woodcock (Scolopax rusticola). Sci Total Environ 363:126–135

    Article  Google Scholar 

  • Chen C, Zhou Q, Liu S, Xiu Z (2011) Acute toxicity, biochemical and gene expression responses of the earthworm Eisenia fetida exposed to polycyclic musks. Chemosphere 83(8):1147–1154

    Article  CAS  Google Scholar 

  • Christofoletti C, Francisco A, Fontanetti CS (2012) Biosolid soil application: toxicity tests under laboratory conditions. Appl Environ Soil Sci. doi: 10.1155/2012/518206.

  • Clauss W (2001) Epithelial transport and osmoregulation in annelids. Can J Zool 79:192–203

    Article  CAS  Google Scholar 

  • Contreras-Ramos SM, Escamilla–Silva EM, Dendooven L (2005) Vermicomposting of biosolids with cow dung and oat straw. Biol Fertil Soils 41:190–198

    Article  Google Scholar 

  • Creamer RE, Rimmer DL, Black HIJ (2008) Do elevated soil concentrations of metals affect the diversity and activity of soil invertebrates in the long-term? Soil Use Manage 24:37–46

    Article  Google Scholar 

  • Del Rio D, Stewart AJ, Pellegrini N (2005) A review of recent studies on malondialdehyde as toxic molecule and biological markers of oxidative stress. Nutr Metab Cardiovasc Dis 15(4):316–328

    Article  Google Scholar 

  • Directive 86/278/EEC (1986) On the protection of the environment, and in particular of the soil, when sewage sludge is used in agriculture. Official Journal of the European Communities No. 181:6–12

    Google Scholar 

  • Directive 91/271/EEC (1991) Concerning urban wastewater treatment. Official Journal of the European Communities No. L 135:40–52

    Google Scholar 

  • Domene X, Solà L, Ramírez W, Alcañiz JM, Andrés P (2011) Soil bioassays as tools for sludge compost quality assessment. Waste Manag 31(3):512–522

    Article  CAS  Google Scholar 

  • Domínguez-Crespo MA, Sánchez-Hernández ZE, Torres-Huerta AM, Negrete-Rodríguez MX, Conde-Barajas E, Flores-Vela A (2012) Effect of the heavy metals Cu, Ni, Cd and Zn on the growth and reproduction of epigeic earthworms (E. fetida) during the vermistabilization of municipal sewage sludge. Water Air Soil Pollut 223:915–931

    Article  Google Scholar 

  • Egiarte G, Pinto M, Ruíz-Romera E, Camps Arbestain M (2008) Monitoring heavy metal concentrations in leachates from a forest soil subjected to repeated applications of sewage sludge. Environ Pollut 156:840–848

    Article  CAS  Google Scholar 

  • EPA (2012) 850.3100 ecological effects test guidelines—earthworm subchronic toxicity test. Environmental Protection Agency (Office of Chemical Safety and Pollution Prevention), US

    Google Scholar 

  • Epel D, Luckenbach T, Stevenson CN, Macmanus-Spencer LA, Hamdoun A, Smital T (2008) Efflux transporters: newly appreciated roles in protection against pollutants. Environ Sci Technol 42:3914–3920

    Article  CAS  Google Scholar 

  • Fuentes A, Lloréns M, Sáez J, Soler A, Aguilar M, Ortuño F (2004) Phytotoxicity and heavy metals speciation of stabilized sewage sludges. J Hazard Mater 108:161–169

    Article  CAS  Google Scholar 

  • García-Gómez C, Fernández MD, Babin M (2014) Ecotoxicological evaluation of sewage sludge contaminated with zinc oxide nanoparticles. Arch Environ Contam Toxicol 67(4):494–506

    Article  Google Scholar 

  • Hait S, Tare V (2012) Transformation and availability of nutrients and heavy metals during integrated composting–vermicomposting of sewage sludges. Ecotoxicol Environ Saf 79:214–224

    Article  CAS  Google Scholar 

  • Han Y, Zhu L, Wang J, Wang J, Xie H, Zhang S (2014) Integrated assessment of oxidative stress and DNA damage in earthworms (Eisenia fetida) exposed to azoxystrobin. Ecotoxicol Environ Saf 107:214–219

    Article  CAS  Google Scholar 

  • He C, Chen CL, Giannis A, Yang Y, Wang JY (2014) Hydrothermal gasification of sewage sludge and model compounds for renewable hydrogen production: a review. Renew Sustain Energy Rev 39:1127–1142

    Article  CAS  Google Scholar 

  • Huang K, Li F, Fu X, Chen X (2013) Feasibility of a novel vermitechnology using vermicast as substrate for activated sludge disposal by two epigeic earthworm species. Agr Sci 4(10):529–535

    CAS  Google Scholar 

  • ISO (1993) Soil quality—effects of pollutants on earthworms (Eisenia fetida). 11268–1 determination of acute toxicity using artificial soil substrate. International Organization for Standardization, Geneva

    Google Scholar 

  • ISO (1998) Soil quality—effects of pollutants on earthworms (Eisenia fetida). 11268–2 determination of effects on reproduction. International Organization for Standardization, Geneva

  • Kinney CA, Furlong ET, Koplin DW, Burkhardt MR, Zaugg SD, Werner SL, Bossio JP, Benotti MJ (2008) Bioaccumulation of pharmaceuticals and other anthropogenic waste indicators in earthworms from agricultural soil amended with biosolid or swine manure. Environ Sci Technol 42(6):1863–1870

    Article  CAS  Google Scholar 

  • Kinney CA, Campbell BR, Thompson R, Furlong ET, Kolpin DW, Burkhardt MR, Zaugg SD, Werner SL, Hay AG (2012) Earthworm bioassays and seedling emergence for monitoring toxicity, aging and bioaccumulation of anthropogenic waste indicator compounds in biosolids-amended soil. Sci Total Environ 433:507–515

    Article  CAS  Google Scholar 

  • Kizilkaya R, Hepşen Türkay FŞ (2014) Vermicomposting of anaerobically digested sewage sludge with hazelnut husk and cow manure by earthworm Eisenia foetida. Compost Sci Util 22:68–82

    Article  CAS  Google Scholar 

  • Kotowska U, Kapelewska J, Sturgulewska J (2014) Determination of phenols and pharmaceuticals in municipal wastewaters from Polish treatment plants by ultrasound-assisted emulsification-microextraction followed by GC-MS. Environ Sci Pollut Res 21:660–673

    Article  CAS  Google Scholar 

  • Kurelec B (1997) A new type of hazardous chemicals: the chemosensitizers of multixenobiotic resistance. Environ Health Perspect 105:855–860

    Article  CAS  Google Scholar 

  • Latare AM, Kumar O, Singh SK, Gupta A (2014) Direct and residual effect of sewage sludge on yield, heavy metals content and soil fertility under rice-wheat system. Ecol Eng 69:17–24

    Article  Google Scholar 

  • LeBlanc RJ, Matthews P, Roland PR (2008) Global atlas of excreta wastewater sludge, and biosolids management: moving forward the sustainable and welcome uses of a global resource. Kenya, United Nations Human Settlements Programme (UN-HABITAT)

    Google Scholar 

  • Li X, Xing M, Yang J, Dai X (2014) Earthworm eco-physiological characteristics and quantification of earthworm feeding in vermifiltration system for sewage sludge stabilization using stable isotopic natural abundance. J Hazard Mater 276:353–361

    Article  CAS  Google Scholar 

  • Litman T, Druley TE, Stein WD, Bates SE (2001) From MDR to MXR: new understanding of multidrug resistance systems, their properties and clinical significance. Cell Mol Life Sci 58(7):931–959

    Article  CAS  Google Scholar 

  • Liu J, Lu Z, Yang J, Xing M, Yu F, Guo M (2012) Effect of earthworms on the performance and microbial communities of excess sludge treatment process in vermifilter. Bioresour Technol 117:214–221

    Article  CAS  Google Scholar 

  • Liu F, Zhu P, Sheng W, Xue JJ (2013) Sludge earthworm composting technology by Eisenia fetida. J Mater Cycles Waste Manag 15:482–488

    Article  CAS  Google Scholar 

  • Lourenço J, Silva A, Carvalho F, Oliveira J, Malta M, Mendo S, Gonçalves F, Pereira R (2011) Histopathological changes in the earthworm Eisenia andrei associated with the exposure to metals and radionuclides. Chemosphere 85(10):1630–1634

    Article  Google Scholar 

  • Ludibeth SM, Marina IE, Vicenta EM (2012) Vermicomposting of sewage sludge: earthworm population and agronomic advantages. Compost Sci Util 20(1):11–17

    Article  Google Scholar 

  • Malara A, Oleszczuk P (2013) Application of a battery of biotests for the determination of leachate toxicity to bacteria and invertebrates from sewage sludge-amended soil. Environ Sci Pollut Res 20:3435–3446

    Article  CAS  Google Scholar 

  • Mann KH (1961) Leeches (Hirudinea): their structure, physiology, ecology and embryology. Pergamon, Oxford

    Google Scholar 

  • McBride MB (2003) Toxic metals in sewage sludge-amended soils: has promotion of beneficial use discounted the risks? Adv Environ Res 8:5–19

    Article  CAS  Google Scholar 

  • Mininni G, Blanch AR, Lucena F, Berselli S (2015) EU policy on sewage sludge utilization and perspectives on new approaches of sludge management. Environ Sci Pollut Res 22:7361–7374

    Article  CAS  Google Scholar 

  • Ministry of Environmental Protection, Physical Planning and Construction (2008) Directive on management of sludge from wastewater treatment plants when the sludge is intended for agricultural use. Official Gazette No. 38/08

    Google Scholar 

  • Molina MJ, Soriano MD, Ingelmo F, Llinares J (2013) Stabilisation of sewage sludge and vinasse bio-wastes by vermicomposting with rabbit manure using Eisenia fetida. Biresour Technol 137:88–97

    Article  CAS  Google Scholar 

  • Nahmani J, Hodson ME, Black S (2007) A review of studies performed to assess metal uptake by earthworms. Environ Pollut 145:402–424

    Article  CAS  Google Scholar 

  • Nogarol LR, Fontanetti CS (2010) Acute and subchronic exposure of diplopods to substrate containing sewage mud: tissular responses of the midgut. Micron 41:239–246

    Article  CAS  Google Scholar 

  • OECD (1984) Guidelines for testing of chemicals—section 2. Effects on biotic systems. Earthworm, acute toxicity tests (filter paper test and artificial soil test). Organisation for Economic Cooperation and Development, Paris

    Google Scholar 

  • OECD (2004) Guideline for testing of chemicals. Earthworm reproduction test (Eisenia fetida/Eisenia andrei). Organisation for Economic Cooperation and Development, Paris

    Book  Google Scholar 

  • Pathak A, Dastidar MG, Sreekrishnan TR (2009) Bioleaching of heavy metals from sewage sludge: a review. J Environ Manage 90:2343–2353

    Article  CAS  Google Scholar 

  • Perez DGP, Fontanetti CS (2010) Assessment of the toxic potential of sewage sludge in the midgut of the diplopod Rhinocricus padbergi. Water Air Soil Pollut 218:437–444

    Article  Google Scholar 

  • Poleksić V, Lenhardt M, Jarić I, Djordjević D, Gačić Z, Cvijanović G, Rašković B (2010) Liver, gills, and skin histopathology and heavy metal content of the Danube sterlet (Acipenser ruthenus Linnaeus, 1758). Environ Toxicol Chem 29(3):515–521

    Article  Google Scholar 

  • Reinecke AJ, Reinecke SA (2004) Earthworm as test organisms in ecotoxicological assessment of toxicant impacts on ecosystems. In: Edwards CA (ed) Earthworm Ecology, 2nd edn. CRC, USA, pp 299–320

    Chapter  Google Scholar 

  • Ribera D, Narbonne JF, Arnaud C, Saint-Denis M (2001) Biochemical responses of the earthworm Eisenia fetida andrei exposed to contaminated artificial soil, effects of carbaryl. Soil Biol Biochem 33:1123–1130

    Article  CAS  Google Scholar 

  • Sauerborn Klobučar R, Žaja R, Franjević D, Brozović A, Smital T (2010) Presence of ecotoxicologically relevant P-gp and MRP transcripts and proteins in cyprinid fish. Arh Hig Rada Toksikol 61:175–182

    Google Scholar 

  • Schaefer M (2003) Behavioral endpoints in earthworm ecotoxicology: evaluation of different test systems in soil toxicity assessment. J Soil Sediment 3(2):79–84

    Article  CAS  Google Scholar 

  • Schreck E, Geret F, Gontier L, Treilhou M (2008) Neurotoxic effect and metabolic responses induced by a mixture of six pesticides on the earthworm Aporrectodea caliginosa nocturna. Chemosphere 71(10):1832–1839

    Article  CAS  Google Scholar 

  • Shaheen SM, Shams MS, Ibrahim SM, Elbehiry FA, Antoniadis V, Hooda PS (2014) Stabilization of sewage sludge by using various by-products: effects on soil properties, biomass production, and bioavailability of copper and zinc. Water Air Soil Pollut 225. doi: 10.1007/s11270-014-2014-x

  • Sharma VJ, Satyanarayan S (2011) Effect of selected heavy metals on the histopathology of different tissues of earthworm Eudrillus eugeniae. Environ Monit Assess 180(1–4):257–267

    Article  CAS  Google Scholar 

  • Silveira MLA, Alleoni LRF, Guilherme LRG (2003) Review: biosolids and heavy metals in soils. Sci Agric 60:793–806

    Article  CAS  Google Scholar 

  • Singh RP, Agrawal M (2008) Potential benefits and risks of land application of sewage sludge. Waste Manag 28:347–358

    Article  CAS  Google Scholar 

  • Smital T, Sauerborn R, Hackenberger BK (2003) Inducibility of the P-glycoprotein transport activity in the marine mussel Mytilus galloprovincialis and the freshwater mussel Dreissena polymorpha. Aquat Toxicol 65:443–465

    Article  CAS  Google Scholar 

  • Sohal RS, Mockett RJ, Orr WC (2002) Mechanisms of aging: an appraisal of the oxidative stress hypothesis. Free Radic Biol Med 33:575–586

    Article  CAS  Google Scholar 

  • Topić Popović N, Strunjak-Perović I, Sauerborn Klobučar R, Barišić J, Babić S, Jadan M, Kepec S, Kazazić SP, Matijatko V, Beer Ljubić B, Car I, Klobučar GV, Čož-Rakovac R (2015) Impact of treated wastewater effluent on fish and toxicity biosensors. Sci Total Environ 538:23–37

    Article  Google Scholar 

  • Wang Y, Wu S, Chen L, Wu C, Yu R, Wang Q, Zhao X (2012) Toxicity assessment of 45 pesticides to the epigeic earthworm Eisenia fetida. Chemosphere 88:484–491

    Article  CAS  Google Scholar 

  • Xing M, Yang J, Wang Y, Liu J, Yu F (2011) A comparative study of synchronous treatment of sewage and sludge by two vermifiltrations using an epigeic earthworm Eisenia fetida. J Hazard Mater 185:881–888

    Article  CAS  Google Scholar 

  • Xing M, Li X, Yang J, Lv B, Lu Y (2012) Performance and mechanism of vermifiltration system for liquid-state sewage sludge treatment using molecular and stable isotopic techniques. Chem Eng J 197:143–150

    Article  CAS  Google Scholar 

  • Xing M, Lv B, Zhao C, Yang J (2014) Towards understanding the effects of additives on the vermicomposting of sewage sludge. Environ Sci Pollut Res 22:4644–4653

    Article  Google Scholar 

  • Yadav A, Garg VK (2011) Recycling of organic wastes by employing Eisenia fetida. Bioresour Technol 102:2874–2880

    Article  CAS  Google Scholar 

  • Žaja R, Klobučar GIV, Sauerborn Klobučar R, Hackenberger BH, Smital T (2006) Haemolymph as compartment for efficient and non-destructive determination of P-glycoprotein (Pgp) mediated MXR activity in bivalves. Comp Biochem Physiol C Toxicol Pharmacol 143(1):103–112

    Article  Google Scholar 

  • Žaja R, Terzić S, Senta I, Lončar J, Popović M, Ahel M, Smital T (2013) Identification of P-glycoprotein inhibitors in contaminated freshwater sediments. Environ Sci Technol 47(9):4813–4821

    Article  Google Scholar 

  • Zhang Q, Zhang B, Wang C (2014) Ecotoxicological effects on the earthworm Eisenia fetida following exposure to soil contaminated with imidacloprid. Environ Sci Pollut Res 21:12345–12353

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The financial support by the Ministry of Science, Education and Sport of the Republic of Croatia (project No. 098-1782739-2749) is acknowledged. The sampling was carried out as a part of doctoral thesis titled “Ecotoxicological and microbiological characterization of purified wastewater of the city Virovitica”. Special thanks are due to the PhD student Slavko Kepec.

Author information

Authors and Affiliations

  1. Division of Materials Chemistry, Ruđer Bošković Institute, Zagreb, Croatia

    S. Babić, J. Barišić, O. Malev, N. Topić Popović, I. Strunjak-Perović, R. Čož-Rakovac & R. Sauerborn Klobučar

  2. Division of Zoology, Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia

    G. Klobučar

  3. Division for Marine and Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia

    N. Krasnići

  4. PathCon Laboratories EU, Prilaz Ivana Visina 1-3, 10000, Zagreb, Croatia

    R. Sauerborn Klobučar

Authors
  1. S. Babić
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Barišić
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. O. Malev
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Klobučar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. Topić Popović
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. I. Strunjak-Perović
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. N. Krasnići
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R. Čož-Rakovac
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R. Sauerborn Klobučar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R. Sauerborn Klobučar.

Additional information

Responsible editor: Thomas Braunbeck

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Babić, S., Barišić, J., Malev, O. et al. Sewage sludge toxicity assessment using earthworm Eisenia fetida: can biochemical and histopathological analysis provide fast and accurate insight?. Environ Sci Pollut Res 23, 12150–12163 (2016). https://doi.org/10.1007/s11356-016-6097-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-016-6097-3

Keywords

Search

Navigation