Prevalence of antibiotic-resistant fecal bacteria in a river impacted by both an antibiotic production plant and urban treated discharges.

In this study, the abundance and spatial dynamics of antibiotic-resistant fecal bacteria (Escherichia coli, total coliforms and Enterococcus spp.) were determined in water and sediment samples from a river impacted by both antibiotic production plant (APP) and urban wastewater treatment plant (WWTP) discharges. Agar dilution and disk diffusion methods were also used for antimicrobial susceptibility testing. Two antimicrobial agents, cephalexin (25 µg/ml) and amoxicillin (50 µg/ml), were evaluated using the agar dilution method for E. coli, total coliforms (TC) and Enterococcus spp., whereas the degree of sensitivity or resistance of E. coli isolates to penicillin (10 U), ampicillin (10 µg), doxycycline (30 µg), tetracycline (30 µg), erythromycin (15 µg), azithromycin (15 µg) and streptomycin (10 µg) was performed using the disk diffusion method. Real-time PCR assays were used to determine the prevalence of three antibiotic-resistance genes (ARGs). The agar dilution method showed that most E. coli isolates and TC were resistant to amoxicillin, especially after receiving the APP discharges. Antibiotic resistances to amoxicillin and cephalexin were higher after the APP discharge point than after the WWTP effluent. The disk diffusion method revealed that 100% of bacterial isolates were resistant to penicillin and erythromycin. Multidrug-resistant bacteria were detected and showed a higher proportion at the WWTP discharge point than those in the APP. Highly multidrug-resistant bacteria (resistance to more than 4 antibiotics) were also detected, reaching mean values of 41.6% in water samples and 50.1% in sediments. The relative abundance of the blaTEM, blaCTX-M and blaSHV genes was higher in samples from the treatment plants than in those collected upstream from the discharges, especially for water samples collected at the APP discharge point. These results clearly demonstrate that both the APP and the WWTP contribute to the emergence and spread of antibiotic resistance in the environment.

Effect of climatic conditions, season and wastewater quality on contaminant removal efficiency of two experimental constructed wetlands in different regions of Spain.

The aim of this study was to examine the effects of climate, season and wastewater quality on contaminant removal efficiency of constructed wetlands implemented in Mediterranean and continental-Mediterranean climate region of Spain. To this end, two experimental horizontal subsurface flow constructed wetlands located in Barcelona and León (Spain) were compared. The two constructed wetland systems had the same experimental set-up. Each wetland had a surface area of 2.95 m(2), a water depth of 25 cm and a granular medium of D(60)=7.3 mm, and was planted with Phragmites australis. Both systems were designed in order to operate with a maximum organic loading rate of 6 g(DBO) m(-2) d(-1). Experimental systems operated with a hydraulic loading rate of 28.5 and 98 mm d(-1) in Barcelona and León, respectively. Total suspended solids, biochemical oxygen demand and ammonium mass removal efficiencies followed seasonal trends, with higher values in the summer (97.4% vs. 97.8%; 97.1% vs. 96.2%; 99.9% vs. 88.9%, in Barcelona and León systems, respectively) than in the winter (83.5% vs. 74.4%; 73.2% vs. 60.6%; 19% vs. no net removal for ammonium in Barcelona and León systems, respectively). During the cold season, biochemical oxygen demand and ammonium removal were significantly higher in Barcelona system than in León, as a result of higher temperature and redox potential in Barcelona. During the warm season, statistical differences were observed only for ammonium removal. Results showed that horizontal subsurface flow constructed wetland is a successful technology for both regions considered, even if winter seemed to be a critical period for ammonium removal in continental climate regions.

Comparison of interannual removal variation of various constructed wetland types.

Seven mesocosm-scale (1m(2)) constructed wetlands (CWs) of different configurations were operated outdoors for thirty-nine months under the same conditions to assess their ability to remove organic matter and nutrients from urban wastewaters. CWs differed in some design parameters, namely the presence of plants, the species chosen (i.e., Typha angustifolia or Phragmites australis), the flow configuration (i.e., surface flow or subsurface flow) and the presence/absence of a gravel bed. It was observed that, in general, removal efficiencies decreased with the aging of the system and that seasonality had a great influence on CWs. A comparison was made in order to figure out which kind of CW was more efficient for the removal of every pollutant in the long term. Planted systems were clearly better than unplanted systems even in winter. Efficiency differences among CWs were not extremely great, especially after a few years. However, some types of CWs were more adequate for the removal of certain pollutants. The effect of the aging on the main parameters involved in pollutant removal in CWs (temperature, pH, conductivity, dissolved oxygen concentration and redox potential) was assessed. The efficiency of CWs should not be evaluated based on short monitoring periods (1-2 years) after the start-up of the systems, but on longer periods.

Temporal evolution in PPCP removal from urban wastewater by constructed wetlands of different configuration: a medium-term study.

Pharmaceuticals and personal care products (PPCPs) are widely distributed in urban wastewaters and can be removed to some extent by constructed wetlands (CWs). The medium-term (3-5 years) behaviour of these systems regarding PPCP removal is still unknown. Seven mesocosm-scale (1 m(2)) CWs of different configurations were operated outdoors for 39 months under the same conditions to assess their PPCP removal ability and temporal evolution. CWs differed in some design parameters, namely plant presence, species chosen (Typha angustifolia vs Phragmites australis), flow configuration and presence/absence of gravel bed (floating macrophytes surface flow, FM-SF; free-water surface flow, FW-SF; free-water subsurface flow, FW-SSF; or conventional horizontal subsurface flow, SSF). PPCP efficiencies decreased throughout time and performance differences among CWs disappeared with the systems aging. This could be due to a homogenization process in the systems caused by detrimental factors like saturation, clogging and shading. Winter efficiencies were lower than summer ones for salicylic acid, caffeine, methyl dihydrojasmonate, galaxolide and tonalide, and seasonal biological activities seem key factors to explain this fact. Maximal removal efficiencies were achieved in an unplanted-FW-SSF for ketoprofen (47-81%), naproxen (58-81%) and salicylic acid (76-98%); in an unplanted-SSF for caffeine (65-99%); in a Phragmites-FM-SF for ibuprofen (49-96%) and diclofenac (16-68%); in a Typha-FM-SF for carbamazepine (35-71%); and in a Typha-FW-SSF for methyl dihydrojasmonate (71-96%), galaxolide (67-82%) and tonalide (55-74%). Photodegradation could be involved in ketoprofen, naproxen, ibuprofen and diclofenac removal. Carbamazepine and diclofenac were moderately removed by the most efficient CWs studied. Carbamazepine might be eliminated by vegetal uptake.

Removal of antibiotics from urban wastewater by constructed wetland optimization.

Seven mesocosm-scale constructed wetlands (CWs), differing in their design characteristics, were set up in the open air to assess their efficiency to remove antibiotics from urban raw wastewater. A conventional wastewater treatment plant (WWTP) was simultaneously monitored. The experiment took place in autumn. An analytical methodology including HPLC-MS/MS was developed to measure antibiotic concentrations in the soluble water fraction, in the suspended solids fraction and in the WWTP sludge. Considering the soluble water fraction, the only easily eliminated antibiotics in the WWTP were doxycycline (61±38%) and sulfamethoxazole (60±26%). All the studied types of CWs were efficient for the removal of sulfamethoxazole (59±30-87±41%), as found in the WWTP, and, in addition, they removed trimethoprim (65±21-96±29%). The elimination of other antibiotics in CWs was limited by the specific system-configuration: amoxicillin (45±15%) was only eliminated by a free-water (FW) subsurface flow (SSF) CW planted with Typha angustifolia; doxycycline was removed in FW systems planted with T. angustifolia (65±34-75±40%), in a Phragmites australis-floating macrophytes system (62±31%) and in conventional horizontal SSF-systems (71±39%); clarithromycin was partially eliminated by an unplanted FW-SSF system (50±18%); erythromycin could only be removed by a P. australis-horizontal SSF system (64±30%); and ampicillin was eliminated by a T. angustifolia-floating macrophytes system (29±4%). Lincomycin was not removed by any of the systems (WWTP or CWs). The presence or absence of plants, the vegetal species (T. angustifolia or P. australis), the flow type and the CW design characteristics regulated the specific removal mechanisms. Therefore, CWs are not an overall solution to remove antibiotics from urban wastewater during cold seasons. However, more studies are needed to assess their ability in warmer periods and to determine the behaviour of full-scale systems.

Statistical modelling of organic matter and emerging pollutants removal in constructed wetlands.

Multiple regression models, clustering tree diagrams, regression trees (CHAID) and redundancy analysis (RDA) were applied to the study of the removal of organic matter and pharmaceuticals and personal care products (PPCPs) from urban wastewater by means of constructed wetlands (CWs). These four statistical analyses pointed out the importance of physico-chemical parameters, plant presence and chemical structure in the elimination of most pollutants. Temperature, pH values, dissolved oxygen concentration, redox potential and conductivity were related to the removal of the studied substances. Plant presence (Typha angustifolia and Phragmites australis) enhanced the removal of organic matter and some PPCPs. Multiple regression equations and CHAID trees provided numerical estimations of pollutant removal efficiencies in CWs. These models were validated and they could be a useful and interesting tool for the quick estimation of removal efficiencies in already working CWs and for the design of new systems which must fulfil certain quality requirements.

Optimization of performance assessment and design characteristics in constructed wetlands for the removal of organic matter.

Some of the most used constructed wetland (CW) configurations [conventional and modified free-water (FW) flow, surface flow, conventional horizontal subsurface flow (SSF) and soilless systems with floating macrophytes (FM)] were assessed in order to compare their efficiencies for the removal of organic pollutants [COD, filtered COD (FCOD), BOD and total suspended solids (TSS)] from urban sewage under the same climatic and wastewater conditions. The removal performance was calculated using three approaches: effluent concentrations, areal removed loads and mass removal. Results were very different depending on the approach, which indicates that the way to present CW efficiency should be considered carefully. All CW-configurations obtained BOD effluent concentrations below 25 mg L(-1) in summer, with a FW-CW with effluent leaving through the bottom of the tank being the only one maintaining low BOD effluent concentrations even in winter and under high organic loading conditions. In this kind of CW, the presence of plants favoured pollutant removal. SSF-CWs were the most efficient for the removal of COD. FM systems can be as efficient as some gravel bed CWs. Typhaangustifolia worked better than Phragmitesaustralis, at least when the systems were at the beginning of their operation period.

Comprehensive assessment of the design configuration of constructed wetlands for the removal of pharmaceuticals and personal care products from urban wastewaters.

Seven mesocosm-scale constructed wetlands (CWs) of different configurations were operated outdoors for nine months to assess their ability to remove pharmaceuticals and personal care products (PPCPs) from urban wastewaters. CWs differed in some design parameters, namely the presence of plants, the species chosen (i.e., Typha angustifolia vs Phragmites australis), flow configuration (i.e., surface flow vs subsurface flow) and the presence of a gravel bed. A nearby conventional activated-sludge wastewater treatment plant (WWTP) fed with the same sewage was simultaneously monitored for comparison. The PPCPs ketoprofen, naproxen, ibuprofen, diclofenac, salicylic acid, carbamazepine, caffeine, galaxolide, tonalide and methyl dihydrojasmonate were monitored. The presence of plants favoured the removal of some PPCPs. The performance of the mesocosm studied was compound-dependant, soilless CWs showing the highest removal efficiency for ketoprofen, ibuprofen and carbamazepine, while free-water CWs with effluent leaving through the bottom of the tank performed well for the degradation of ketoprofen, salicylic acid, galaxolide and tonalide. Finally, subsurface horizontal flow CWs were efficient for the removal of caffeine. Significant linear correlations were observed between the removal of some PPCPs and temperature or redox potential. Hence, microbiological pathways appear to be the most probable degradation route for PPCPs in the CWs studied.