Effect of ciprofloxacin antibiotic on the partial-nitritation process and bacterial community structure of a submerged biofilter.

A partial-nitritation bench-scale submerged biofilter was used for the treatment of synthetic wastewater containing a high concentration of ammonium in order to study the influence of the antibiotic ciprofloxacin on the partial-nitritation process and biodiversity of the bacterial community structure. The influence of ciprofloxacin was evaluated in four partial-nitritation bioreactors working in parallel, which received sterile synthetic wastewater amended with 350 ng/L of ciprofloxacin (Experiment 1), synthetic wastewater without ciprofloxacin (Experiment 2), synthetic wastewater amended with 100 ng/L of ciprofloxacin (Experiment 3) and synthetic wastewater amended with 350 ng/L of ciprofloxacin (Experiment 4). The concentration of 100 ng/L of antibiotics demonstrated that the partial-nitritation process, microbial biomass and bacterial structure generated by tag-pyrosequencing adapted progressively to the conditions in the bioreactor. However, high concentrations of ciprofloxacin (350 ng/L) induced a decay of the partial-nitritation process, while the total microbial biomass was increased. Within the same experiment, the bacterial community experienced sequential shifts with a clear reduction of the ammonium oxidation bacteria (AOB) and an evident increase of Commamonas sp., which have been previously reported to be ciprofloxacin-resistant. Our study suggests the need for careful monitoring of the concentration of antibiotics such as ciprofloxacin in partial-nitritation bioreactors, in order to choose and maintain the most appropriate conditions for the proper operation of the system.

Influence of sludge retention time and temperature on the sludge removal in a submerged membrane bioreactor: comparative study between pure oxygen and air to supply aerobic conditions.

Performance of a bench-scale wastewater treatment plant, which consisted of a membrane bioreactor, was monitored daily using pure oxygen and air to supply aerobic conditions with the aim of studying the increases of the aeration and sludge removal efficiencies and the effect of the temperature. The results showed the capacity of membrane bioreactor systems for removing organic matter. The alpha-factors of the aeration were determined for six different MLSS concentrations in order to understand the system working when pure oxygen and air were used to supply aerobic conditions in the system. Aeration efficiency was increased between 30.7 and 45.9% when pure oxygen was used in the operation conditions (a hydraulic retention time of 12 h and MLSS concentrations between 4,018 and 11,192 mg/L). Sludge removal efficiency increased incrementally, from 0.2 to 1.5% when pure oxygen was used at low sludge retention time and from 1.5% to 15.4% at medium sludge retention time when temperature conditions were lower than 20°C. Moreover, the difference between calculated and experimental sludge retention time was lesser when pure oxygen was used to provide aerobic conditions, so the influence of the temperature decreased when the pure oxygen was used. These results showed the convenience of using pure oxygen due to the improvement in the performance of the system.

Removal and degradation characteristics of quinolone antibiotics in laboratory-scale activated sludge reactors under aerobic, nitrifying and anoxic conditions.

This work describes the removal of 6 quinolone antibiotics from wastewaters under different redox conditions (aerobic, nitrifying and anoxic) through batch experiments in laboratory scale activated sludge reactors using mixed liquor from a membrane bioreactor pilot plant (MBR). The main removal pathways for antibiotics from wastewaters involved in each treatment are described. Mass balances indicated that sorption on sludge played a dominating role in the elimination of antibiotics. Sorption potential depended on the redox conditions, being lower in nitrifying (Kd, 414-876 L kg(-1)) and anoxic (Kd, 471-930 L kg(-1)) sludge in comparison with aerobic sludge (Kd, 534-1137 L kg(-1)). Kd was higher for piperazinylic quinolones. Redox conditions also influenced biodegradation, a secondary pathway, which followed first-order kinetics with degradation rates constants ranging from 1.8·10(-3) to 8.2·10(-3) h(-1). Biodegradation rates under anoxic conditions were negligible. The experimental results have also demonstrated much higher removal efficiency by biodegradation (36.2-60.0%) under nitrifying conditions in comparison with aerobic conditions (14.9-43.8%). The addition of allylthiourea, an ammonia monooxygenase inhibitor, inhibited nitrification completely and reduced significantly the biodegradation of target antibiotics (16.5-29.3%). The residual biodegradation in the presence of allylthiourea may be due to the activity of heterotrophs in the enriched nitrifier culture. The removal of the selected antibiotics under the studied redox conditions depended significantly on the bacteria composition of the sludge. These results suggest that despite the known persistence of this group of antibiotics it is possible to enhance their degradation using nitrifying conditions, which at adequate working conditions as high SRT, typical in MBR, become a promising alternative for improving quinolones removal from environment.

Biological and technical study of a partial-SHARON reactor at laboratory scale: effect of hydraulic retention time.

This study was on the technical and biological characteristics of a partial-SHARON submerged-filter bioreactor of 3 L. The main focus was the influence of the hydraulic retention time (HRT) on biofilms. For this purpose, we used molecular tools based on the partial 16S rRNA genes. The results showed that the HRT may affect the nitrification processes of a bioreactor using synthetic wastewater containing 600 mg/L of ammonia. It was found that an HRT of 0.5 day transformed 100 % of the ammonium into nitrite. However, when the HRT was decreased to 0.4 day, there was a significant reduction (35 %) in the quantity of ammonia transformed, which confirmed the complexity of the system operation. Moreover, a PCR-TGGE approach highlighted the differences observed. The results obtained showed that an HRT of 0.5 day reduced bacterial biodiversity in the biofilms, which were mainly formed by Nitrosomonas and Diaphorobacter. In contrast, an HRT of 0.4 day facilitated the formation of heterogeneous biofilms formed by nitrifying bacteria, such as Nitrosomonas sp., Nitrosospira sp., and Nitrosovibrio sp.).

Influence of filling ratio and carrier type on organic matter removal in a moving bed biofilm reactor with pretreatment of electrocoagulation in wastewater treatment.

At present, there is great concern about limited water resources and water quality, which require a more advanced technology. The Moving Bed Biofilm Reactor (MBBR) has been shown to be an efficient technology for removal of organic matter and nutrients in industrial and urban wastewater treatment. However, there are some pollutants which are more difficult to remove by biological processes, so this process can be improved with additional physical and chemical treatments such as electrocoagulation, which appears to be a promising technology in electrochemical treatments. In this research, urban wastewater was treated in an MBBR plant with an electrocoagulation pre-treatment. K1 from AnoxKaldnes and AQWISE ABC5 from Aqwise were the carriers studied under three different filling ratios (20, 35, and 50%). The experimental pilot plant had four bioreactors with 20 L of operation volume and a common feed tank with 100 L of operation volume. The movement of the carriers was generated by aeration and stirrer systems. Organic matter removal was studied by analysis of soluble chemical oxygen demand (sCOD). The maximum organic matter removal in this MBBR system was 65.8% ± 1.4% and 78.4% ± 0.1% for K1 and Aqwise ABC5 carriers, respectively. Moreover, the bacterial diversity of the biofilm was studied by temperature-gradient gel electrophoresis (TGGE) of PCR-amplified partial 16S rRNA genes. 20 prominent TGGE bands were successfully reamplified and sequenced, being the predominant population: ß-Proteobacteria, α-Proteobacteria, and Actinobacteria.

Treatment of effluents polluted by nitrogen with new biological technologies based on autotrophic nitrification-denitrification processes.

In recent years, various technologies have been developed for the removal of nitrogen from wastewater that is rich in nitrogen but poor in organic carbon, such as the effluents from anaerobic digesters and from certain industries. These technologies have resulted in several patents. The core of these technologies is some of the processes and patents described in this paper: Aerobic denitrification, Sharon, Anammox, OLAND, CANON, NOx process, DEMON. More specifically, one of the first innovative options described for removing nitrogen include partial nitrification under aerobic conditions (partial Sharon process) followed by autotrophic anaerobic oxidation (Anammox process). The partial Sharon-Anammox process can be performed under alternating oxic and anoxic conditions in the same bioreactor or in two steps in two separate bioreactors. This overview focuses on the technical and biological aspects of these new types of treatment system, and compares them to other technologies. Given the fact that nitrification is a sensitive process, special attention is paid to conditions such as temperature, dissolved oxygen, hydraulic retention time, free ammonia, nitrous acid concentration, and pH. A discussion of the pros and cons of such treatment systems is also included since autotrophic nitrogen removal has advantages as well as drawbacks. The paper concludes with a discussion of future research that could improve these systems by enhancing performance and reducing costs.

Advanced methods for the elimination of microorganisms in industrial treatments: potential applicability to wastewater reuse.

Because of the growing need to eliminate undesirable microorganisms in different industrial treatments, mainly in the food and agricultural sector and the pharmaceutical industry, a number of increasingly effective systems for disinfection to eliminate microorganisms have been devised. This article analyzes different methods to eliminate and/or significantly reduce the number of microorganisms in industrial contexts and in environmental engineering. Although, in the past, thermal treatments had been used most frequently for microbial elimination, the method is costly and has the disadvantage of modifying the organoleptic and/or physicochemical properties of the food products. For this reason, new technologies rapidly are being developed, such as high-intensity pulsed electric fields, high-pressure systems, ultrasounds, and irradiation, which effectively eliminate microorganisms without deteriorating the properties of the product. These emerging technologies are potentially applicable in the field of environmental engineering.

Performance of bench-scale membrane bioreactor under real work conditions using pure oxygen: viscosity and oxygen transfer analysis.

Pure oxygen to supply the aerobic condition was used in the performance of a bench-scale submerged membrane bioreactor (MBR). The pilot plant was located in the wastewater treatment plant of the city of Granada (Spain) and the experimental work was divided into two stages (Unsteady state and steady state conditions). Operation parameters (MLSS, MLVSS and dissolved oxygen concentration) and physical characteristics (temperature, conductivity, pH, COD and BOD(5)) were daily monitored. The results showed the capacity of the MBR systems to remove organic material under a hydraulic retention time of 18.46 h and sludge retention time of 18.6 days. Therefore, Viscosity of the sludge and alphakLa-factor of the aeration, were determinate in the steady stage condition to understand the behavior of the system when pure oxygen has been used to supply the aerobic conditions of the MBR system showed an alpha-factor of 0.238 when the viscosity of the system was 4.04 Cp.

Structure of archaeal communities in membrane-bioreactor and submerged-biofilter wastewater treatment plants.

A cultivation independent approach (PCR-TGGE) was used to evaluate the occurrence of Archaea in four wastewater treatments based on technologies other than activated sludge, and to comparatively analyze their community structure. TGGE fingerprints (based on partial archaeal 16S-rRNA amplicons) were obtained from sludge samples taken from a pilot-scale aerated MBR fed with urban wastewater and operated under two different sets of conditions (MBR1 and MBR2 treatments), and also from biofilms sampled from two pilot-scale submerged biofilters (SBs) consisting of one aerated and one anoxic column each, fed with urban (USB treatment) or industrial (ISB treatment) wastewater, respectively. Analysis of TGGE fingerprints revealed clear and significant differences of the community structure of Archaea between the wastewater treatments studied, primarily according to wastewater origin and the type of technology. Thirty-two different band classes were detected among the 23 sludge and biofilm samples analyzed, from which five were selected as dominant or distinctive of the four treatments studied. Sixteen predominant TGGE bands were identified, revealing that all of them were related to methanogenic Archaea. Neither other Euryarchaeota groups nor Crenarchaeota members were identified amongst the 16S-rRNA fragments sequenced from separated TGGE bands.

Microbial enzymatic activities in a pilot-scale MBR experimental plant under different working conditions.

Phosphatases, glucosidase, protease, esterase and dehydrogenase activities in a MBR (membrane bioreactor) system equipped with ultrafiltration membranes for the treatment of real urban wastewater were measured at different volatile suspended solid (VSS) concentrations, total suspended solid (TSS) concentrations, hydraulic retention times (HRT), temperatures and inflow rates. The results showed the capacity of the MBR system to remove COD and BOD(5) at TSS between 7200 and 13,300 mg/L; HRT values of 8.05 and 15.27 h; inflow rates of 14.67 and 27.81 L/h; and temperatures between 4 and 27 degrees C. The enzymatic activities are influenced by increases in VSS and TSS concentrations. These results suggest that the ability to get adapted to environmental changes of the bacterial populations and their microbial enzymatic activities is essential to understand the biological processes that occur in MBR systems and crucial for proper urban wastewater treatment when using MBR technologies.

Microbial community structure and dynamics in a pilot-scale submerged membrane bioreactor aerobically treating domestic wastewater under real operation conditions.

A pilot scale submerged ultra-filtration membrane bioreactor (MBR) was used for the aerobic treatment of domestic wastewater over 9 months of year 2006 (28th March to 21st December). The MBR was installed at a municipal wastewater facility (EMASAGRA, Granada, Spain) and was fed with real wastewater. The experimental work was divided in 4 stages run under different sets of operation conditions. Operation parameters (total and volatile suspended solids, dissolved oxygen concentration) and environmental variables (temperature, pH, COD and BOD(5) of influent water) were daily monitored. In all the experiments conducted, the MBR generated an effluent of optimal quality complying with the requirements of the European Law (91/271/CEE 1991). A cultivation-independent approach (polymerase chain reaction-temperature gradient gel electrophoresis, PCR-TGGE) was used to analyze changes in the structure of the bacterial communities in the sludge. Cluster analysis of TGGE profiles demonstrated significant differences in community structure related to variations of the operation parameters and environmental factors. Canonical correspondence analysis (CCA) suggested that temperature, hydraulic retention time and concentration of volatile suspended solids were the factors mostly influencing community structure. 23 prominent TGGE bands were successfully reamplified and sequenced, allowing gaining insight into the identities of predominantly present bacterial populations in the sludge. Retrieved partial 16S-rRNA gene sequences were mostly related to the alpha-Proteobacteria, beta-Proteobacteria and gamma-Proteobacteria classes. The community established in the MBR in each of the four stages of operation significantly differed in species composition and the sludge generated displayed dissimilar rates of mineralization, but these differences did not influence the performance of the bioreactor (quality of the permeate). These data indicate that the flexibility of the bacterial community in the sludge and its ability to get adapted to environmental changes play an important role for the stable performance of MBRs.

Effect of the concentration of suspended solids on the enzymatic activities and biodiversity of a submerged membrane bioreactor for aerobic treatment of domestic wastewater.

A pilot-scale submerged membrane bioreactor was used for the treatment of domestic wastewater in order to study the influence of the variations in the concentration of volatile suspended solids (VSS) on the enzymatic activities (acid and alkaline phosphatases, glucosidase, protease, esterase, and dehydrogenase) and biodiversity of the bacterial community in the sludge. The influence of VSS concentration was evaluated in two separated experiments, which were carried out in two different seasons of the year (experiment 1 through spring-summer and experiment 2 through autumn-winter). Cluster analysis of the temperature gradient gel electrophoresis (TGGE) profiles demonstrated that the community composition was significantly different in both experiments. Within the same experiment, the bacterial community experienced sequential shifts as the biomass accumulated, as shown by the evolution of the population profiles through time as VSS concentration increased. All enzymatic activities studied were significantly lower during experiment 2, except for glucosidase. Concentrations of VSS over 8 g/l induced a strong descent of all enzymatic activities, which overlapped with a significant modification of the community composition. Sequences of the major TGGE bands were identified as representatives of the Alpha-proteobacteria, filamentous bacteria (Thiotrix), and nitrite oxidizers (Nitrospira). Some sequences which were poorly related to any validated bacterial taxon were obtained.

Biological nitrogen and phenol removal from saline industrial wastewater by submerged fixed-film reactor.

In this study a biological nitrogen removal process using a submerged fixed-film reactor was applied to treat industrial wastewater with phenol (1g/l), a high nitrogen concentration (0.4 g N/l) and high salinity (30 g/l). The process consisted of a pre-denitrification system with a down-flow-up-flow biofilter (two columns, each with an effective volume of 21 l) packed with clayey schists from recycled construction material. The efficiency of the system for reducing COD, phenol concentration and total nitrogen was tested under different running conditions such as influent flow (10, 12 and 15 l/d), air loading (6.8 and 13.6m(3)/m(2)h) and effluent recirculation (300%, 400%, and 600%). The system demonstrated a high capacity for reducing COD concentration (95.75+/-0.72%), independently of running conditions. The aerobic column eliminated most of the phenol in the influent. Nitrogen removal took place mainly in the anoxic column, and was conditioned by the air loading in the aerated column, owing to the dependence of nitrification on the supply of oxygen. However, this process was not able to achieve a nitrogen oxidation superior to 63%, in spite of a sufficient supply of oxygen and the diluting effect of high recirculation (600%) on the phenol concentration in the influent. In spite of the limitations observed in the process of nitrification, results for the removal of total nitrogen were as high as 83%, owing to a combination of different processes for nitrogen removal.

Treatment of landfill leachate with aerated and non-aerated submerged biofilters.

A submerged biological filter packed with clayey schists from brick industrial waste as support material was used to treat landfill leachate. The pilot-scale plant consisted of two separated reactors, running in aerated and non-aerated conditions respectively. The leachate was taken from a landfill in Granada (southeast Spain) and was considered as partially stabilized with average chemical oxygen demand (COD) concentrations of 18,683.3 mg COD/L. The efficiency of the system at reducing COD, biological oxygen demand (BOD(5)), total suspended solids (TSS), total volatile solids (TVS), total solids (TS), cations and anions concentrations was tested comparing aerating and non-aerating conditions under several volumic organic loadings (15.7, 25.1 and 31.1 kg COD/m(3)d), a hydraulic loading of 0.7 m(3)/m(2)d and an internal recirculation of 250%. Results obtained showed that the aerated reactor was more effective than the non-aerated reactor in the removal of COD, BOD5 and TS, with maximum efficiencies of 66.7%, 91.2% and 21.7%, respectively. The volumic organic loading of 25.1 COD/m(3)d obtained best results. However, for the removal of TSS and TVS the non-aerated reactor showed better results (42%). The behaviour of different cations (Na(+), K(+), and Mg(2+)) and anions (F(-), Cl(-), Br(-)) was also monitored and a reduction in their concentration in the effluent was detected, in the order of 61% for Ca(2+) and 100% for F(-).

Analysis of community composition of biofilms in a submerged filter system for the removal of ammonia and phenol from industrial wastewater.

The bacterial diversity of a submerged filter, used for the removal of ammonia and phenol from an industrial wastewater with high salinity, was studied by a cultivation-independent approach based on PCR/TGGE (temperature-gradient gel electrophoresis). The wastewater treatment plant (laboratory scale) combined the nitrification and denitrification processes and consisted of two separated columns (one anoxic and one aerated) connected through a valve. The spatial diversity of bacterial communities in the plant biofilms was analysed by taking samples at four different heights in the system. TGGE profiles of PCR-amplified sequences of the 16 S rRNA gene (V3-hypervariable region) showed significant variations of the bacterial diversity, mainly depending on the concentration of O(2) along the system. Several bands separated by TGGE were reamplified and sequenced, in order to explore the composition of the microbial communities in the biofilms. Most of the sequenced bands (10 out of 13) were closely related to the 16 S rRNA gene of marine alpha-proteobacteria, mainly grouping in the periphery of the genus Roseobacter. Other sequences were related to those of gamma-proteobacteria, the nitrite oxidizer Nitrospira marina and anaerobic phenol-degrading bacteria of the Desulfobacteraceae.

Coagulation-flocculation pretreatment of a partially stabilized leachate from a sanitary landfill site at Alhendin (Granada, Southern Spain).

Leachate recirculation, biological, and physicochemical treatment processes are used to treat leachate landfill, but all treatment technologies seem to need a combination of two or more methods to obtain an effluent with suitable properties to eliminate environmental problems. In spite of the considerable literature focused on the coagulation-flocculation applied to landfill leachate before biological treatments, most investigations are developed to study different operating variables as leachate characteristics, type, and concentrations of chemical products applied. Two ferric coagulants (ferric chloride and Ferriclar), two aluminum coagulants (PAX-18 and SAL-8.2), an organic coagulant and flocculant (Chemifloc PA-15), several anionic flocculant (Chemifloc A05.L, A10.L, A20L, and A30.L), and a non-ionic flocculant (Chemifloc N01) were used in jar-test experiments to determine the best conditions for the removal of chemical oxygen demand (COD) and total solids (TS) from a partially stabilized leachate collected at a sanitary landfill site at Alhendín (Granada, Southern Spain). The organic coagulant-flocculant PA-15 showed highest (17%) COD and TS removal efficiencies at a concentration of 2 mL/L. All combinations of coagulants and flocculants enhanced the COD removal efficiency. The best results (more than 25% COD removal) were obtained with a combination of ferric or aluminum-based salts in combination with high molecular weight flocculants.

Influence of inocula over start up of a denitrifying submerged filter applied to nitrate contaminated groundwater treatment.

Pure culture of denitrifying bacteria isolated from heterogeneous biofilm was applied as inocula to a submerged filter for removing nitrogen from contaminated groundwater. Five highly denitrifying bacteria were used, comparing their attachment, nitrogen-removal ability and final water quality in the start up of the system. Our experiments showed that inocula selection is the crucial step when the submerged filter is applied to obtain drinking water, since selected strains vary in their ability to colonise support material and to remove nitrogen, and in their effect on treated water quality during start up phase. Hidrogenophaga pseudoflava strain proved to be the most suitable inoculum out of the five tested, under the experimental conditions.

Wastewater treatment using fibrist and saprist peat: a comparative study.

Two different peat types (fibrist and saprist) were compared to examine their respective wastewater treatment capacity in peat beds under different hydraulic loadings (1.8, 1.2, 0.9, and 0.6 m3/m2 d) and pollutant loadings (dependent on influent quality). The comparative study was carried out on a pilot scale using urban wastewater which had undergone preliminary treatment. The systems were observed to retain a higher quantity of suspended solids when the load was higher, regardless of the hydraulic loading and the type of peat employed. System performance with respect to BOD was similar to that observed for COD, and it was noted that the capacity to retain organic matter decreased when the hydraulic loading or pollutant loading was increased. This effect was observed in both peats but to different degrees, being lower in the case of saprist peat. The results indicate that it would be necessary to work with low hydraulic loadings in order to comply with legislative requirements on effluent. Peat beds are therefore suitable for implementation in small population centers, where the system's efficiency partly depends on the type of peat employed. In view of the system's performance with respect to suspended solids, this technology may be considered a good primary treatment, and could be usefully combined with other processes aimed at eliminating dissolved organic matter.

Inoculation of a submerged filter for biological denitrification of nitrate polluted groundwater: a comparative study.

Activated sludge from a wastewater treatment plant and pure culture of Hydrogenophaga pseudoflava were utilized for the development of a denitrifying biofilm in a submerged filter in order to remove nitrate from polluted groundwater. Nitrate removal efficiency, nitrite accumulation, turbidity, COD and faecal indicators persistence in the treated water were determined at different superficial hydraulic loading (10, 20 and 30 m(3)/m(2) d) and superficial nitrate loading rates (1, 2, 3, 6 and 9 Kg NO(3)(-)/m(2) d) in the submerged filter. The application of H. pseudoflava as inocula allowed better results in terms of system stability, higher superficial hydraulic loading and superficial nitrate loading rates (30 m(3)/m(2)d and 9 kg NO(3)(-) /m(2) d, respectively). These values improve those obtained when the system was inoculated with activated sludge. In addition, the pure microbial inocula improved design parameters and running of the process due to its biofilm homogeneity, obtaining treated water with better characteristics to its final use as drinking water than that obtained with an activated sludge inocula.

Influence of hydraulic loading and air flowrate on urban wastewater nitrogen removal with a submerged fixed-film reactor.

Nutrient disposal to sensitive areas, particularly nitrogen and phosphorus from wastewater treatment plants, provokes eutrophication reducing water quality. Fixed film technology is widely used for the removal of organic matter and nitrogen by the biological process of nitrification-denitrification. This paper studies a nitrification and post-denitrification lab-scale plant with a downflow aerobic submerged filter for removal of organic matter and nitrification, followed by an anoxic upflow biofilter for denitrification. Recycled construction material (clay shists) was employed as support material and methanol was used as carbon source. After 2 weeks of acclimation in which nitrification reached steady-state conditions, different hydraulic loadings (0.35-1.59 m(3)/m(2)h) and air flowrates (7.78-43.5 m(3)/m(2)h) were applied for 1 year. The highest hydraulic loading which complied with the EU regulation on nitrogen disposal was 0.71 m(3)/m(2)h (1.6 h). Hydraulic retention time (HRT), which corresponded to a nitrogen removal of 0.64 kg N/m(3) per day operating at an air flowrate of 25.6 m(3)/m(2)h. Concerning to organic matter removal efficiency, the aerobic reactor accepted a maximum chemical oxygen demand (COD) volumetric loading of 16.0 kg COD/m(3) per day with a 75% COD removal efficiency. For all the tests carried out, suspended solids (SS) concentration in the outlet water was less than 35 mg/l.