Ciprofloxacin degradation in anaerobic sulfate-reducing bacteria (SRB) sludge system: Mechanism and pathways.

Ciprofloxacin (CIP), a fluoroquinolone antibiotic, removal was examined for the first time, in an anaerobic sulfate-reducing bacteria (SRB) sludge system. About 28.0% of CIP was biodegraded by SRB sludge when the influent CIP concentration was 5000 μg/L. Some SRB genera with high tolerance to CIP (i.e. Desulfobacter), were enriched at CIP concentration of 5000 μg/L. The changes in antibiotic resistance genes (ARGs) of SRB sludge coupled with CIP biodegradation intermediates were used to understand the mechanism of CIP biodegradation for the first time. The percentage of efflux pump genes associated with ARGs increased, while the percentage of fluoroquinolone resistance genes that inhibit the DNA copy of bacteria decreased during prolonged exposure to CIP. It implies that some intracellular CIP was extruded into extracellular environment of microbial cells via efflux pump genes to reduce fluoroquinolone resistance genes accumulation caused by exposure to CIP. Additionally, the degradation products and the possible pathways of CIP biodegradation were also examined using the new method developed in this study. The results suggest that CIP was biodegraded intracellularly via desethylation reaction in piperazinyl ring and hydroxylation reaction catalyzed by cytochrome P450 enzymes. This study provides an insight into the mechanism and pathways of CIP biodegradation by SRB sludge, and opens-up a new opportunity for the treatment of CIP-containing wastewater using sulfur-mediated biological process.

Jia Y ，Khanal SK ，Shu H ，Zhang H ，Chen GH ，Lu H ... -  《-》

Stress-responses of activated sludge and anaerobic sulfate-reducing bacteria sludge under long-term ciprofloxacin exposure.

The activated sludge (AS) and sulfate-reducing bacteria (SRB) sludge systems were continuously operated for 200 days in laboratory to investigate the stress-responses of these two sludge systems under ciprofloxacin (CIP) exposure. It was found that CIP was effectively removed by SRB sludge via adsorption and biodegradation, but little biodegradation in AS system. The CIP biodegradation by SRB sludge made the SRB sludge system more sustainable and tolerant to long-term CIP exposure than AS system with significant (p < 0.05) CIP desorption and decrease of CIP removal. CIP shaped the microbial communities in AS and SRB sludge, and significantly (p < 0.05) inhibited the family Nitrosomonadaceae (ammonia-oxidizing bacteria (AOB)) and genus Nitrospira (nitrite-oxidizing bacteria (NOB)/complete ammonia oxidizer(comammox)) and the nitrogen removal in AS system. Moreover, CIP posed the increase of genus Zoogloea-like organisms and the non-filamentous bulking of AS, e.g. 313 ± 12 mL/g of sludge volume index (SVI) at phase V (influent CIP = 5000 μg/L). The genus Desulfobacter was enriched in SRB sludge system under long-term CIP exposure, and stimulated chemical oxygen demand (COD) removal and sulfate reduction. The increase of genera Zoogloea, Acinetobacter and Flavobacterium in AS, and Zoogloea and Acinetobacter in SRB sludge systems under CIP exposure promoted extracellular polymeric substances (EPS) production and CIP adsorption for self-protection of microbes against CIP toxicity. The functional groups of N-H, O-H, C-O-C and C=O in EPS of AS and SRB sludge provided adsorption sites for CIP and impeded CIP impact on microbial cells. The findings of this study provide an insight into the stress-responses of AS and SRB sludge under long-term CIP exposure, and exhibit the great potential of treating CIP-laden wastewater by SRB sludge system.

Zhang H ，Song S ，Jia Y ，Wu D ，Lu H ... -  《-》

Insights into pharmaceuticals removal in an anaerobic sulfate-reducing bacteria sludge system.

In this study, we examined eight typical and widely detected pharmaceuticals (PhAs) removal in an anaerobic sulfate-reducing bacteria (SRB) sludge system (five antibiotics: sulfadiazine (SD), sulfamethoxazole (SMX), trimethoprim (TMP), ciprofloxacin (CIP) and enoxacin (ENO), and three nonsteroidal anti-inflammatory drugs (NSAIDs): ibuprofen (IBU), ketoprofen (KET) and diclofenac (DIC)). The results showed that the SRB sludge had the higher removal efficacy (20 to 90%) for antibiotics (SD, SMX, TMP, CIP and ENO) than NSAIDs (<20%) via adsorption and biodegradation under different operating conditions. Based on a series of batch studies, fluoroquinolone antibiotics (CIP and ENO) were instantly (<15 min) removed (∼98%) via adsorption on SRB sludge with adsorption coefficient (Kd) as high as 25.3 ± 1.8 L/g-suspended solids (SS). And thermodynamics results indicated that the adsorption of CIP and ENO on SRB sludge was spontaneous (Gibbs free energy change (ΔG°) <0 kJ/mol), exothermic (enthalpy change (ΔH°) <0 kJ/mol), and the adsorption process involved both physisorption and chemisorption (absolute value of ΔH° = 20 to 80 kJ/mol). Three widely prescribed antibiotics (SMX, TMP and CIP) were further investigated for their possible biodegradation pathways along with functional enzymes involved through a series of batch experiments. The biotransformation intermediates indicated that biotransformations of SMX and CIP in SRB sludge system could be initiated from the cleavage of isoxazole and piperazinyl rings catalyzed by sulfite reductase (SR) and cytochrome P450 (CYP450) enzymes, respectively. TMP was likely biotransformed via O-demethylation and N-acetylation coupled with hydroxylation reactions with CYP450 enzymes as the main functional enzymes. This study provided new insight into PhAs removal in SRB sludge system, and has significant potential of implementing sulfur-mediated biological process for the treatment of PhAs containing wastewater.

Jia Y ，Zhang H ，Khanal SK ，Yin L ，Lu H ... -  《-》

Understanding the Role of Extracellular Polymeric Substances on Ciprofloxacin Adsorption in Aerobic Sludge, Anaerobic Sludge, and Sulfate-Reducing Bacteria Sludge Systems.

Zhang H ，Jia Y ，Khanal SK ，Lu H ，Fang H ，Zhao Q ... -  《-》

Ciprofloxacin-degrading Paraclostridium sp. isolated from sulfate-reducing bacteria-enriched sludge: Optimization and mechanism.

Ciprofloxacin (CIP), one of the most widely used fluoroquinolone antibiotics, is frequently detected in the effluents of wastewater treatment plants and aquatic environments. In this study, a CIP-degrading bacterial strain was isolated from the sulfate reducing bacteria (SRB)-enriched sludge, identified as Paraclostridium sp. (i.e., strain S2). The effects of critical operational parameters on CIP removal by the strain S2 were systematically studied and these parameters were optimized via response surface methodology to maximize CIP removal. Furthermore, the pathway and kinetics of CIP removal were investigated by varying the initial CIP concentrations (from 0.1 to 20 mg/L). The CIP removal was characterized by rapid sorption followed by biotransformation with a specific biotransformation rate of 1975.7 ± 109.1 µg/g-cell dry weight/h at an initial CIP concentration of 20 mg/L. Based on the main transformation products, several biotransformation pathways have been proposed including piperazine ring cleavage, OH/F substitution, decarboxylation, and hydroxylation as the major transformation reactions catalyzed by cytochrome P450 and dehydrogenases. Acute toxicity assessment apparently shows that CIP biotransformation by strain S2 resulted in the formation of less toxic intermediates. To the best of our knowledge, this is the very first study in which a key functional microbe, Paraclostridium sp., highly effective in CIP biotransformation, was isolated from SRB-enriched sludge. The findings of this study could facilitate in developing appropriate bioaugmentation strategy, and in designing and operating an SRB-based engineered process for treating CIP-laden wastewater.

Fang H ，Oberoi AS ，He Z ，Khanal SK ，Lu H ... -  《-》