Environmental Science and Pollution Research

, Volume 21, Issue 13, pp 8112–8123 | Cite as

Genotoxic assessment and optimization of pressmud with the help of exotic earthworm Eisenia fetida

  • Sartaj Ahmad Bhat
  • Jaswinder Singh
  • Adarsh Pal VigEmail author
Research Article


Genotoxicity of pressmud (PM) to Allium cepa was investigated to assess its toxic potential and to elucidate the effect of vermicomposting to reduce its toxicity. The PM produced as a waste by product of the sugar cane industry was mixed with cow dung (CD) at different ratios of 0:100 (V0), 25:75 (V25), 50:50 (V50), 75:25 (V75) and 100:0 (V100) (PM:CD) on a dry weight basis for vermicomposting with Eisenia fetida. Different concentrations of 100 % PM sludge extract (10 %, 20 %, 40 %, 60 %, 80 % and 100 %) and negative control (distilled water) and positive control (maleic hydrazide) were analyzed with A. cepa assay to evaluate frequency of chromosomal aberrations before and after vermicomposting. Percent aberration was greatest (30.8 %) after exposure to 100 % PM extract after 6 h but was reduced to 20.3 % after vermicomposting. Exposure to the extract induced c-mitosis, delayed anaphase, laggards, stickiness and vagrant aberrations. Microscopic examination of root meristem exposed to PM sludge extract showed significant inhibition of mitotic index. Also, the mitotic index decreased with increase in concentration of PM sludge extract. After vermicomposting the mitotic index was increased. However, increasing percentages of PM significantly affected the growth and fecundity of the worms and maximum population size was reached in the 25:75 (PM:CD) feed mixture. Nitrogen, phosphorus, sodium, electrical conductivity (EC) and pH increased from initial feed mixture to the final products (i.e., vermicompost), while organic carbon, C/N ratio and potassium declined in all products of vermicomposting. Scanning electron microscopy (SEM) was recorded to identify the changes in texture with numerous surface irregularities and high porosity that proves to be good vermicompost manure. It could be concluded that vermicomposting could be an important tool to reduce the toxicity of PM as evidenced by the results of genotoxicity.


Vermicomposting Genotoxicity Industrial waste Pressmud Earthworm Allium cepa test Chromosomal abnormalities 



This work was financially supported by Department of Botanical and Environmental Science, Guru Nanak Dev University, Amritsar, Punjab, India.


  1. Aira M, Monroy F, Dominguez J (2007) Earthworms strongly modify microbial biomass and activity triggering enzymatic activities during vermicomposting independently of the application rates of pig slurry. Sci Total Environ 385:252–261CrossRefGoogle Scholar
  2. APHA (1998) Standard methods for examination of water and wastewater. 17th ed. American Public Health Association, Washington, DCGoogle Scholar
  3. Arillo A, Melodia F (1991) Reduction of hexavalent chromium by the earthworm E. fetida (Savigny). Ecotoxicol Environ Saf 57:391–394Google Scholar
  4. Atiyeh RM, Arancon NQ, Edwards CA, Metzger JD (2001) The influence of earthworms Processed pig manure on the growth and productivity of marigold. Bioresour Technol 81:103–108CrossRefGoogle Scholar
  5. Bhat SA, Singh J, Vig AP (2013) Vermiremediation of dyeing sludge from textile mill with the help of exotic earthworm Eisenia fetida Savigny. Environ Sci Pollut Res 20:5975–5982CrossRefGoogle Scholar
  6. Bremner JM, Mulvaney CS (1982) Nitrogen total. In: Page AL, Miller RH, Keeney DR (eds) Methods of soil analysis. Am. Soc. Agronomy, Madison, WI, pp 575–624Google Scholar
  7. Deolalikar AV, Mitra A, Bhattacharyee S, Chakraborty S (2005) Effect of vermicomposting process on metal content of paper mill solid waste. J Environ Sci Eng 47:81–84Google Scholar
  8. Dixit GB, Nerle SK (1985) Cytotoxic effect of industrial effluents on Allium cepa L. Geophys J Roy Astron Soc 12:240–273Google Scholar
  9. Edwards CA, Dominguez J, Neuhauser EF (1998) Growth and reproduction of Perionyx excavatus (Perr.) (Megascolecidae) as factors in organic waste management. Biol Fertil Soils 27:155–161CrossRefGoogle Scholar
  10. Fernandes TCC, Mazzeo DEC, Marin-Morales MA (2007) Mechanism of micronuclei formation in polyploidizated cells of Allium cepa exposed to trifluralin herbicide. Pest Biochem Physiol 88:252–259CrossRefGoogle Scholar
  11. Ferrari B, Radetski CM, Veber AM, Ferard JF (1999) Ecotoxicological assessment of solid waste: a combined liquid and solid phase testing approach using a battery of bioassays and biomarkers. Environ Toxicol Chem 18:1195–1202Google Scholar
  12. Fischer E, Koszorus L (1992) Sublethal, accumulation capacities and elimination rates of As, Hg and Se in the manure worms Eisenia fetida. Pedobiologia 36:172–178Google Scholar
  13. Fiskesjo G (1985) The Allium test as a standard for environmental monitoring. Hereditas 102:99–112CrossRefGoogle Scholar
  14. Fiskesjo G (1988) The Allium test—an alternative in environmental studies: the relative toxicity of metal ions. Mutat Res 197:243–260CrossRefGoogle Scholar
  15. Hoshina MM (2002) Avaliação da possível contaminação das águas do Ribeirão Claro — município de Rio Claro, pertencente à bacia do rio Corumbataí, por meio de testes de mutagenicidade em Allium cepa, Trabalho de conclusão (Bacharel e Licenciatura - Ciências Biológicas). Universidade Estadual Paulista, Rio Claro/SP, p 52Google Scholar
  16. Jain K, Singh J, Chauhan LKS, Murthy RC, Gupta SK (2004) Modulation of flyash-induced genotoxicity in Vicia faba by vermicomposting. Ecotoxicol Environ Saf 59:89–94CrossRefGoogle Scholar
  17. Jiang W, Liu D (2000) Effects of Pb on root growth, cell division, and nucleolus of Zea mays L. Bull Environ Contam Toxicol 65:786–793CrossRefGoogle Scholar
  18. John MK (1970) Colorimetric determination of phosphorus in soil and plant material with ascorbic acid. Soil Sci 109:214–220CrossRefGoogle Scholar
  19. Joshi BD, Singh TP (1989) Cellular and chromosomal effect of tannery effluent in A. cepa meristem XVI Annual conference EMSI, Abst. 29Google Scholar
  20. Kaur A, Singh J, Vig AP, Dhaliwal SS, Rup PJ (2010) Cocomposting with and without Eisenia fetida for conversion of toxic paper mill sludge into soil conditioner. Bioresour Technol 101:8192–8198CrossRefGoogle Scholar
  21. Kaviraj B, Sharma S (2003) Municipal solid waste management through vermicomposting employing exotic and local species of earthworms. Bioresour Technol 90:169–173CrossRefGoogle Scholar
  22. Knasmuller S, Gotlmann E, Steinkellner H, Fomin A, Pickle C, Paschke A, God R, Kundi M (1998) Detection of genotoxic effects of heavy metal contaminated soils with plant bioassays. Mutat Res 420:37–38CrossRefGoogle Scholar
  23. Masood F, Malik A (2013) Mutagenicity and genotoxicity assessment of industrial wastewaters. Envi Sci Pollut Res 20:7386–7397CrossRefGoogle Scholar
  24. Nagao M (1978) Environmental mutagens and carcinogens. Annu Rev Gent 12:117–159CrossRefGoogle Scholar
  25. Nelson DW, Sommers LE (1996) Total carbon and organic carbon and organic matter. In: Page AL, Miller RH, Keeney DR (eds) Method of soil analysis. Am. Soc. of Agronomy, Madison, WI, pp 539–579Google Scholar
  26. Neuhauser EF, Hartenstein R, Kaplan DL (1980) Growth of the earthworm Eisenia foetida in relation to population density and food rationing. OIKOS 35:93–98CrossRefGoogle Scholar
  27. Odeigah PGC, Nurudeen O, Amund OO (1997) Genotoxicity of oil field wastewater in Nigeria. Hereditas 126:161–167CrossRefGoogle Scholar
  28. Plaza C, Nogales R, Senesi N, Benitez E, Polo A (2007) Organic matter humification by vermicomposting of cattle manure alone and mixed with two phase olive pomace. Bioresour Technol 9:5085–5089Google Scholar
  29. Prakash M, Karmegam N (2010) Vermistabilization of pressmud using Perionyx ceylanensis. Mich Bioresour Technol 101:8464–8468CrossRefGoogle Scholar
  30. Pramanik P, Ghosh GK, Ghosal PK, Banik P (2007) Changes in organic –C, N, P and K and enzyme activities in vermicompost of biodegradable organic wastes under limiting and microbial inoculants. Bioresour Technol 98:2485–2494CrossRefGoogle Scholar
  31. Qian XW (2004) Mutagenic effects of chromium trioxide on root tips of Vicia faba. J Zhejiang Univ Sci 5:1570–1576CrossRefGoogle Scholar
  32. Rank J, Nielsen MH (1997) Allium cepa anaphase–telophase root tip chromosome aberration assay on N-methyl-N-nitrosourea, maleic hydrazide, sodium azide, and ethyl methanesulfonate. Mutat Res 390:121–127CrossRefGoogle Scholar
  33. Russel PJ (2002) Chromosomal mutation B. Cummings. Genetics, Pearson Education Inc, San Francisco, pp 595–621Google Scholar
  34. Sangwan P, Kaushik CP, Garg VK (2010) Vermicomposting of sugar industry waste (pressmud) mixed with cow dung employing an epigeic earthworm Eisenia foetida. Waste Manag Res 28:71–75CrossRefGoogle Scholar
  35. Sen B, Chandra TS (2007) Chemolytic and solid-state spectroscopic evaluation of organic matter transformation during vermicomposting of sugar industry wastes. Bioresour Technol 98:1680–1683CrossRefGoogle Scholar
  36. Sharma S, Nagpal A, Vig AP (2012) Genoprotective potential of Brassica juncea (L.) Czern. Against mercury-induced genotoxicity in Allium cepa L. Turk J Biol 36:622–629Google Scholar
  37. Singh J, Kaur A, Vig AP, Rup PJ (2010) Role of Eisenia fetida in rapid recycling of nutrients from bio sludge of beverage industry. Ecotoxicol Environ Saf 73:430–435CrossRefGoogle Scholar
  38. Subramanian S, Sivarajan M, Saravanapriya S (2010) Chemical changes during vermicomposting of sago industry solid wastes. J Hazard Mater 179:318–322CrossRefGoogle Scholar
  39. Tripathi G, Bhardwaj P (2004) Comparative studies on biomass production, life cycles and composting efficiency of Eisenia foetida (Savigny) and Lampito mauritii (Kinberg). Bioresour Technol 92:275–278CrossRefGoogle Scholar
  40. Vig AP, Singh J, Wani S, Dhaliwal SS (2011) Vermicomposting of tannery sludge mixed with cattle dung into valuable manure using earthworm Eisenia fetida (Savigny). Bioresour Technol 102:7941–7945CrossRefGoogle Scholar
  41. Yadav A, Garg VK (2011) Recycling of organic wastes by employing Eisenia fetida. Bioresour Technol 102:2874–2880CrossRefGoogle Scholar
  42. Zeyer J, Ranganathan LS, Chandra TS (2004) Pressmud as biofertilizer for improving soil fertility and pulse crop productivity. ISCB — Indo-Swiss collaboration in Biotech. A Report. Portfolia. First Phase (1999–2004)Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Sartaj Ahmad Bhat
    • 1
  • Jaswinder Singh
    • 2
  • Adarsh Pal Vig
    • 1
    Email author
  1. 1.Department of Botanical and Environmental SciencesGuru Nanak Dev UniversityAmritsarIndia
  2. 2.Department of ZoologyKhalsa College AmritsarPunjabIndia

Personalised recommendations