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- Strategies to enhance the removal of FluoroquinolonesPublication . Amorim, Catarina L.; Maia, Alexandra S.; Moreira, Irina S.; van Loosdrecht, Mark C.M.; Tiritan, Maria E.; Castro, Paula M.L.Fluoroquinolones (FQs) are broad-spectrum antibiotics that play an important role in the treatment of serious bacterial infections. Currently, several FQs are available but ciprofloxacin (CPF), ofloxacin (OFL) and norfloxacin (NOR) are amongst the most worldwide prescribed antibiotics. Antibiotics can reach wastewater treatment plants (WWTP) from different routes. Thus removal of these contaminants during the biotreatment process is of major importance in order to avoid their release to other environmental matrices. Granular sludge sequencing batch reactors (SBR) constitute a novel biofilm technology for wastewater treatment extremely promising for the treatment of effluents containing toxic compounds. Therefore, in this study a granular sludge SBR, established with activated sludge from a WWTP, was operated for the treatment of an aqueous stream containing FQs. No evidence of FQ biodegradation followed by HPLC with Fluorescence Detection was observed but FQs adsorbed to the aerobic granular sludge, being gradually released into the medium after withdrawal of the FQs in the inlet stream. In a previous study, Labrys portucalensis F11 demonstrated to be able to degrade FQs, namely OFL, NOR and CPF, when supplied individually or as a mixture, in the presence of an easy degradable carbon source. Different removal extents were obtained for the tested concentrations (ranging from 0.8 to 30 μM), but overall the uptake capacity of strain F11 for individual FQs decreased with increasing the initial FQ concentration. When supplied with a mixture FQs, strain F11 concomitantly removed each target antibiotic but a decrease on the biodegradability of FQs was observed which could be explained by competition mechanisms. The ability of Labrys portucalensis F11 to grow using the readily available carbon source while maintain its ability to degrade FQs reinforce the potential of this strain in bioaugmentation processes. As the indigenous microbial communities in biotreatment processes rarely are able to remove such contaminants, using this promising FQ-degrading strain, bioaugmentation strategies such as inoculation of the degrading strain, as a suspension or immobilized on carrier material, or using a plasmid donor strain carrying the degradative genes, could be assessed to improve FQ removal. Acknowledgments: C.L. Amorim, A.S. Maia and I.S. Moreira wish to acknowledge the research grants from Fundação para a Ciência e Tecnologia (FCT), Portugal (Ref. SFRH/BD/47109/2008, SFRH/BD/86939/2012 and SFRH/BPD/87251/2012, respectively) and Fundo Social Europeu (Programa Operacional Potencial Humano (POPH), Quadro de Referência Estratégico Nacional (QREN))). This work was supported by FCT through the projects PTDC/EBB-EBI/111699/2009 and PEst-OE/EQB/LA0016/2011.
- Aerobic granules in a sequencing batch bioreactor under fluoroquinolone shock loadingsPublication . Amorim, Catarina L.; Maia, Alexandra S.; Mesquita, Raquel B. R.; Rangel, António O. S. S.; Tiritan, Maria E.; Castro, Paula M. L.The growing occurrence of human and veterinary pharmaceuticals in the environment is causing increasing concern. Fluoroquinolones (FQs) are broad-spectrum antibiotics that play an important role in the treatment of serious bacterial infections. Antibiotics can reach wastewater treatment plants (WWTP) from different routes. Domestic effluents are considered the major contributor but effluents from pharmaceutical industries and hospitals are also of great concern. Granular sequencing batch reactors (SBR) constitute a novel biofilm technology for wastewater treatment extremely promising for the treatment of effluents containing toxic compounds. Aerobic granular sludge has several advantages over activated sludge, such as excellent settling properties, high biomass retention, ability to deal with high organic loading rates and to perform simultaneously diverse biological processes, such as Chemical Oxygen Demand (COD), N and P removal. This study focused on the effect of intermittent and alternating feeding of different FQs, namely Ofloxacin (OFL), Norfloxacin (NOR) and Ciprofloxacin (CPF), on bioreactor performance and diversity of the microbial population. Activated sludge from a municipal WWTP was used as the inoculum for the start-up of the SBR. The aerobic granules grew under aerobic conditions and after ca. 3 months of reactor operation stable granules were observed. The FQs affected the granular sludge in terms of morphology, causing a decrease in granule size. The granules started to disintegrate and an increase in the levels of solids in the effluent after exposure to FQs occurred due to wash-out of unstable granules, concomitant with a decrease in the SBR bed volume. The effect of the target fluorinated pharmaceuticals on the main biological processes occurring in the granular sludge SBR, such as nitrification and phosphate removal, was evaluated. Ammonium and nitrite were practically not detected in the treated effluent (maximum concentration of 0.03 and 0.01 mM for NH4+-N and NO2--N, respectively) indicating that neither ammonia oxidizing bacteria (AOB) nor nitrite oxidizing bacteria (NOB) were inhibited by the presence of the FQs, whereas phosphate removal was affected. The phosphate released into the bulk liquid by the phosphate accumulating organisms (PAO) during the anaerobic feeding period was not completely removed and the levels of phosphate in the bioreactor effluent increased. The organic removal, measured by COD, was not markedly affected by FQ shock loads. Changes in the bacterial community from aerobic granules related to FQs shock loadings were examined using denaturing gradient electrophoresis (DGGE) of 16S rRNA. The clustering analysis suggested that samples clustered according to the temporal factor. The gradual succession observed in the bacterial assemblage composition was related with the exposure to FQs. Also, the microbial population present in the aerobic granules was also investigated by culture-dependent methods. Several bacterial isolates belonging to α- and ɣ-branch of the Proteobacteria phylum were retrieved from the granules. After ca. 2 months without FQs exposure, the SBR bed volume was recovered and the solid content at the bioreactor effluent returned to normal levels.
- Performance of aerobic granular sludge in a sequencing batch bioreactor exposed to ofloxacin, norfloxacin and ciprofloxacinPublication . Amorim, Catarina L.; Maia, Alexandra S.; Mesquita, Raquel B. R.; Rangel, António O.S.S.; Loosdrecht, Mark C.M. van; Tiritan, Maria Elizabeth; Castro, Paula M.L.A granular sludge sequencing batch reactor (SBR) was operated for 340 days for treating a synthetic wastewater containing fluoroquinolones (FQs), namely ofloxacin, norfloxacin and ciprofloxacin. The SBR was intermittently fed with FQs, at concentrations of 9 and 32 mM. No evidence of FQ biodegradation was observed but the pharmaceutical compounds adsorbed to the aerobic granular sludge, being gradually released into the medium in successive cycles after stopping the FQ feeding. Overall COD removal was not affected during the shock loadings. Activity of ammonia oxidizing bacteria and nitrite oxidizing bacteria did not seemto be inhibited by the presence of FQs (maximum of 0.03 and 0.01 mM for ammonium and nitrite in the effluent, respectively). However, during the FQs feeding, nitrate accumulation up to 1.7 mM was observed at the effluent suggesting that denitrificationwas inhibited. The activity of phosphate accumulating organismswas affected, as indicated by the decrease of P removal capacity during the aerobic phase. Exposure to the FQs also promoted disintegration of the granules leading to an increase of the effluent solid content, nevertheless the solid content at the bioreactor effluent returned to normal levelswithin ca. 1month after removing the FQs in the feed allowing recovery of the bedvolume. Denaturing gradient gel electrophoresis revealed a dynamic bacterial community with gradual changes due to FQs exposure. Bacterial isolates retrieved from the granules predominantly belonged to a- and g-branch of the Proteobacteria phylum.
- Aerobic granular sludge sequencing batch reactor performance under fluorinated pharmaceuticals shock loadingsPublication . Moreira, Irina S.; Amorim, Catarina L.; Ribeiro, Ana R.; Maia, Alexandra S.; Mesquita, Raquel B. R.; Calheiros, Cristina S. C.; Rangel, António O. S. S.; Loosdrecht, Mark C. M.; Tiritan, Maria E.; Castro, Paula M. L.The widespread usage of pharmaceuticals is of increasing concern. Aerobic granular sludge sequencing batch reactors (AGS-SBR) constitute a promising technology for the treatment of wastewaters, however how the removal of carbon and nutrients can be affected by such micropollutants is largely unknown. This study evaluates the impact of different fluorinated drugs (ofloxacin, norfloxacin, ciprofloxacin and fluoxetine) on the performance of an AGS-SBR. During 468 days, a sequence of intermittent, alternating and/or continuous shock loads of pharmaceuticals were apllied to an AGS-SBR and the effects on the main biological processes were evaluated. Here we report on the effect of fluoroquinolones on reactor performnce. The organic removal, measured by COD, was not markedly affected by pharmaceuticals shock loads. Ammonium and nitrite were practically not detected in the bioreactor effluent indicating that the presence of the pharmaceuticals did not inhibit nitrification, whereas accumulation of nitrate in the effluent was observed, indicating that denitrification was affected. Phosphate removal was affected to some extent. There was no evidence of biodegradation whereas adsorption of the target pharmaceuticals to the AGS was observed, which were gradually released into the medium after withdrawal from the inlet stream.