Environmental Science and Pollution Research

, Volume 23, Issue 9, pp 9215–9226 | Cite as

Citric acid- and Tween® 80-assisted phytoremediation of a co-contaminated soil: alfalfa (Medicago sativa L.) performance and remediation potential

  • A. C. Agnello
  • D. Huguenot
  • E. D. van Hullebusch
  • G. Esposito
Research Article


A pot experiment was designed to assess the phytoremediation potential of alfalfa (Medicago sativa L.) in a co-contaminated (i.e., heavy metals and petroleum hydrocarbons) soil and the influence of citric acid and Tween® 80 (polyethylene glycol sorbitan monooleate), applied individually and combined together, for their possible use in chemically assisted phytoremediation. The results showed that alfalfa plants could tolerate and grow in a co-contaminated soil. Over a 90-day experimental time, shoot and root biomass increased and negligible plant mortality occurred. Heavy metals were uptaken by alfalfa to a limited extent, mostly by plant roots, and their concentration in plant tissues were in the following order: Zn > Cu > Pb. Microbial population (alkane-degrading microorganisms) and activity (lipase enzyme) were enhanced in the presence of alfalfa with rhizosphere effects of 9.1 and 1.5, respectively, after 90 days. Soil amendments did not significantly enhance plant metal concentration or total uptake. In contrast, the combination of citric acid and Tween® 80 significantly improved alkane-degrading microorganisms (2.4-fold increase) and lipase activity (5.3-fold increase) in the rhizosphere of amended plants, after 30 days of experiment. This evidence supports a favorable response of alfalfa in terms of tolerance to a co-contaminated soil and improvement of rhizosphere microbial number and activity, additionally enhanced by the joint application of citric acid and Tween® 80, which could be promising for future phytoremediation applications.


Soil remediation Heavy metals Hydrocarbons Alfalfa (Medicago sativa L.) Citric acid Tween® 80 



The authors would like to thank the financial support provided by French Île-de-France region and European Commission (Erasmus Mundus Joint Doctorate Programme ETeCoS3: Environmental Technologies for Contaminated Solids, Soils and Sediments, under the grant agreement FPA n°2010-0009). A.C. Agnello acknowledges the Mediterranean Office for Youth (MOY), which granted a 10-month mobility fellowship in the frame of the MOY labeled programs n°2010/038. Finally, the authors appreciate the contribution of Iwona M’Kenzie Hall, who provided English assistance during the preparation of the manuscript.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • A. C. Agnello
    • 1
    • 2
  • D. Huguenot
    • 1
  • E. D. van Hullebusch
    • 1
  • G. Esposito
    • 2
  1. 1.Université Paris-Est, Laboratoire Géomatériaux et Environnement (EA 4508), UPEMMarne-la-ValléeFrance
  2. 2.Dipartimento di Ingegneria Civile e MeccanicaUniversità degli Studi di Cassino e del Lazio MeridionaleCassinoItaly

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