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Influence of Diclofenac on Activated Sludge Bacterial Communities in Fed-Batch Reactors

Barbara Kraigher*orcid tiny and Ines Mandic-Mulecorcid tiny

University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Chair of Microbiology, Večna pot 111,

1000 Ljubljana, Slovenia


Article history:

Received: 24 June 2019

Accepted: 24 November 2020

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Key words:

activated sludge, pharmaceuticals, diclofenac, bacterial community T-RFLP, Pseudomonas


Research background. The occurrence and environmental toxicity of pharmaceuticals have recently attracted increasing attention. Diclofenac is a highly consumed non-steroidal anti-inflammatory drug, which is often detected in wastewaters, but investigations of its influence on bacteria are scarce. 

Experimental approach. We investigated the influence of this pharmaceutical on bacterial community in activated sludge exposed to increasing concentrations of diclofenac in fed-batch reactors over 41 days. Nitrification activity of the activated sludge was measured and changes in bacterial community structure were followed using culture-independent molecular method (terminal restriction fragment length polymorphism, T-RFLP) and by the cultivation approach. 

Results and conclusions. Nitrification activity was not detectably influenced by addition of diclofenac, while the main change of the bacterial community structure was detected only at the end of incubation (after 41 days) when diclofenac was added to artificial wastewater as the only carbon source. Changes in community composition due to enrichment were also observed using cultivation approach. However, taxonomic affiliation of isolates did not match taxons identified by T-RFLP community profiling. Isolates obtained from inoculum activated sludge belonged to five genera: Comamonas, Arthrobacter, Acinetobacter, Citrobacter and Aeromonas, known for their potential to degrade aromatic compounds. However, only Pseudomonas species were isolated after the last enrichment step on minimal agar plates with diclofenac added as the sole carbon source. 

Novelty and scientific contribution. Our results suggested that the selected recalcitrant and commonly detected pharmaceutical does not strongly influence the sensitive and important nitrification process of wastewater treatment neither. Moreover, the isolated strains obtained after enrichment procedure that were able to grow on minimal agar plates with diclofenac added as the only carbon source could serve as potential model bacteria to study bacterial diclofenac degradation.

*Corresponding author: +38613203410

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