Environmental Science and Pollution Research

, Volume 22, Issue 11, pp 8031–8043 | Cite as

Influence of the natural growth environment on the sensitivity of phototrophic biofilm to herbicide

  • A. PauleEmail author
  • A. Lamy
  • V. Roubeix
  • F. Delmas
  • J. L. Rols
Crop protection: environment, human health, and biodiversity


Ecotoxicological experiments were performed in laboratory-scale microcosms to investigate community-level structural responses of river phototrophic biofilms from different environments to herbicide exposure. Biofilms were initially cultivated on artificial supports placed in situ for 4 weeks at two sites, site M, located in an agricultural watershed basin and site S, located in a forested watershed basin. The biofilms were subsequently transferred to microcosms and, after an acclimatisation phase of 7 days were exposed to alachlor at 10 and 30 μg L−1 for 23 days. Alachlor effects were assessed by a combination of structural parameters, including biomass (ash-free dry mass and chlorophyll a), molecular fingerprinting of the bacterial community (polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE)) and diatom species composition. Alachlor impacted the chlorophyll a and ash-free dry mass levels of phototrophic biofilms previously cultivated at site S. The structural responses of bacterial and diatom communities were difficult to distinguish from changes linked to the microcosm incubation period. Phototrophic biofilms from site S exposed at 30 μg L−1 alachlor were characterised by an increase of Achnanthidium minutissimum (K-z.) Czarnecki abundance, as well as a higher proportion of abnormal frustules. Thus, phototrophic biofilms with different histories, exhibited different responses to alachlor exposure demonstrating the importance of growth environment. These observations also confirm the problem of distinguishing changes induced by the stress of pesticide toxicity from temporal evolution of the community in the microcosm.


Alachlor Phototrophic biofilms Bacterial community Diatoms Growth environment Biological succession Microbial ecotoxicology 



This work was funded by the French National Programme EC2CO-Environmental Microbiology—and by the Midi-Pyrénées Council Programme of the Pyrenean working community. We thank J. Ferriol and D. Dalger for assistance with the DGGE and water chemistry analysis, respectively. We also thank the ‘Association des Agriculteurs d’Auradé’ for the access to site M.

Supplementary material

11356_2014_3429_MOESM1_ESM.doc (1 mb)
Fig. S1 Photographs of the two study sites M and S chosen for in situ production of natural phototrophic biofilm. (DOC 4929 kb)
11356_2014_3429_MOESM2_ESM.doc (3.6 mb)
Fig. S2 Different views of artificial substrates (polyethylene plates, 50 × 100 × 5 mm) used for in situ production and collection of natural phototrophic biofilms (site M on the photography). (DOC 3864 kb)
11356_2014_3429_MOESM3_ESM.doc (104 kb)
Fig. S3 Schema of one microcosm subdivided into three compartments by vertically glass slides. Water recirculation is realised with a submerged pump. Seven polyethylene plates are positioned in the main compartment. (DOC 103 kb)
11356_2014_3429_MOESM4_ESM.doc (167 kb)
Fig. S4 A normal diatom frustule (left) versus an abnormal form (right) of Achnanthidium minutissimum observed in the 30 μg L−1 alachlor-contaminated microcosms at 23 days of incubation during the experiment S. Scale bar, 10 μm. (DOC 167 kb)
11356_2014_3429_MOESM5_ESM.doc (40 kb)
Table S1 Values of the pesticides levels obtained from the previous studies (Debenest 2007, Paule et al. 2009) of the two study sites M and S during the springs of the years 2005, 2006 and 2007. (DOC 3883 kb)
11356_2014_3429_MOESM6_ESM.doc (30 kb)
Table S2 Values of the nitrate (NO3 ) levels obtained from the previous studies (Debenest 2007, Paule et al. 2009) of the two study sites M and S during the springs of the years 2005 and 2007. (DOC 3870 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • A. Paule
    • 1
    • 2
    Email author
  • A. Lamy
    • 1
    • 2
  • V. Roubeix
    • 3
  • F. Delmas
    • 3
  • J. L. Rols
    • 1
    • 2
  1. 1.University de Toulouse, UPS, INP, EcoLab (Laboratoire d’écologie fonctionnelle et environnement)ToulouseFrance
  2. 2.CNRS, EcoLabToulouseFrance
  3. 3.IRSTEA, UR Ecosystèmes Aquatiques et Changements Globaux (EABX)CestasFrance

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