Toxicity assessment of pharmaceutical compounds on mixed culture from activated sludge using respirometric technique: The role of microbial community structure

Abstract

Micropollutants of emerging concern such as pharmaceuticals can significantly affect the performance of secondary biological processes in wastewater treatment plants. The present study is aimed to evaluate the toxicity and inhibition of three pharmaceutical compounds (caffeine, sulfamethoxazole and carbamazepine) on two cultures of microbial consortia enriched from wastewater aerobic activated sludge. One of them was acclimated to pharmaceuticals and the other was non-acclimated as control bioassay. The toxic and inhibitory effects on these cultures were assessed by respirometric tests through the oxygen uptake rate as an indicator of their capacity to degrade a readily available carbon source. Higher values of toxicity and inhibition of pharmaceutical compounds were observed for the control culture as compared to the acclimated one. Sulfamethoxazole and carbamazepine exhibited higher toxicity and inhibition effects than caffeine in both acclimated and control cultures. The microbial diversity of the two cultures was also studied. The composition of microbial community of acclimated and control cultures, was determined by targeting the 16S ribosomal RNA gene. It was observed that Proteobacteria was the most abundant phylum, with Gammaproteobacteria dominating both cultures. Control culture was dominated by Gammaproteobacteria and mostly by the genera Pseudomonas and Sodalis, which belong to common families present in wastewater. Results suggested that the acclimated culture to the three pharmaceuticals was mostly comprised of the extremely multiresistant genera Escherichia-Shigella (38%) of Gammaproteobacteria, resulting to higher resistance as compared to the control culture (Escherichia-Shigella, 7%). Finally, the microbial structure of the microorganisms present in a real bioreactor, which was initially seeded with the acclimated culture and fed in a continuous mode with the selected pharmaceuticals, was also analyzed. The continuous loading of pharmaceuticals in the bioreactor affected its microbial diversity, leading to the dominance of Betaproteobacteria and to the resistant genus Rhizobium of Alphaproteobacteria.

Keywords: Caffeine; Carbamazepine; Inhibition; Multiresistant species; Sulfamethoxazole; Toxicity.