Long-term exposure of activated sludge in chemostats leads to changes in microbial communities composition and enhanced biodegradation of 4-chloroaniline and N-methylpiperazine

Chemosphere. 2020 Mar;242:125102. doi: 10.1016/j.chemosphere.2019.125102. Epub 2019 Oct 22.


Exposure history and adaptation of the inoculum to chemicals have been shown to influence the outcome of ready biodegradability tests. However, there is a lack of information about the mechanisms involved in microbial adaptation and the implication thereof for the tests. In the present study, we investigated the impact of a long-term exposure to N-methylpiperazine (NMP) and 4-chloroaniline (4CA) of an activated sludge microbial community using chemostat systems. The objective was to characterize the influence of adaptation to the chemicals on an enhanced biodegradation testing, following the OECD 310 guideline. Cultures were used to inoculate the enhanced biodegradability tests, in batch, before and after exposure to each chemical independently in chemostat culture. Composition and diversity of the microbial communities were characterised by 16s rRNA gene amplicon sequencing. Using freshly sampled activated sludge, NMP was not degraded within the 28 d frame of the test while 4CA was completely eliminated. However, after one month of exposure, the community exposed to NMP was adapted and could completely degrade it. This result was in complete contrast with that from the culture exposed for 3 months to 4CA. Long term incubation in the chemostat system led to a progressive loss of the initial biodegradation capacity of the community, as a consequence of the loss of key degrading microorganisms. This study highlights the potential of chemostat systems to induce adaptation to a specific chemical, ultimately resulting in its biodegradation. At the same time, one should be critical of these observations as the dynamics of a microbial community are difficult to maintain in chemostat, as the loss of 4CA biodegradation capacity demonstrates.

Keywords: 4-chloroaniline; Biodegradability testing; Continuous culture; Microbial community; N-methylpiperazine; Organic pollutants.

MeSH terms

  • Aniline Compounds / metabolism*
  • Biodegradation, Environmental*
  • Microbiota / drug effects*
  • Piperazine / metabolism*
  • RNA, Ribosomal, 16S
  • Sewage / microbiology*
  • Time Factors


  • Aniline Compounds
  • RNA, Ribosomal, 16S
  • Sewage
  • Piperazine
  • 4-chloroaniline