Vancomycin sorption on activated sludge Gram+ bacteria rather than on EPS; 3D Confocal Laser Scanning Microscopy time-lapse imaging.

Antibiotics-bacteria interactions depend on antibiotic concentration at the scale of bacteria. This study investigates how vancomycin penetrates into activated sludge flocs and can be sorbed on the bacteria and extracellular polymeric substances (EPS). The 3D structure of flocs was imaged using EPS autofluorescence. The green fluorescent BODIPY® FL vancomycin was introduced in a microscopic chamber containing activated sludge and penetration of vancomycin into the flocs by diffusion was observed using time-lapse microscopy. The penetration depended on the floc structure, as long and large pores could go through the whole flocs making preferential path. The antibiotic concentration into the flocs was also found to depend on the sorption rate. BODIPY® FL vancomycin was found to bind preferentially into Gram+ bacteria than on EPS. The vancomycin adsorption constant on bacteria according to the linear adsorption model, Kdbacteria was estimated to be 5 times higher (SD 2.6) than the adsorption constant on EPS KdEPS. These results suggest that antibiotic removal by sorption into wastewater treatment plants could change according to the amount of bacteria in the sludge. Moreover, antibiotic concentration at the scale of bacteria could be significantly higher than the concentration in the bulk solution and this should be taken into account when studying antibiotic activity or biodegradation.

A protocol for the use of computational fluid dynamics as a supportive tool for wastewater treatment plant modelling.

To date, computational fluid dynamics (CFD) models have been primarily used for evaluation of hydraulic problems at wastewater treatment plants (WWTPs). A potentially more powerful use, however, is to simulate integrated physical, chemical and/or biological processes involved in WWTP unit processes on a spatial scale and to use the gathered knowledge to accelerate improvement in plant models for everyday use, that is, design and optimized operation. Evolving improvements in computer speed and memory and improved software for implementing CFD, as well as for integrated processes, has allowed for broader usage of this tool for understanding, troubleshooting, and optimal design of WWTP unit processes. This paper proposes a protocol for an alternative use of CFD in process modelling, as a way to gain insight into complex systems leading to improved modelling approaches used in combination with the IWA activated sludge models and other kinetic models.

Spectrophotometric characterization of human impacted surface waters in the Moselle watershed.

In order to characterize the pollution discharged into the Moselle River and some of its tributaries, spectroscopic techniques, namely UV-vis spectroscopy and synchronous fluorescence spectroscopy, have been combined. UV-visible spectra were analysed using the maximum of the second derivative at 225 nm (related to nitrates), the SUVA254 and E2/E3 indices (related to the nature of organic matter). Synchronous fluorescence spectra (delta lambda = 50 nm) presented different shapes depending upon the type of pollution. The pollution results from anthropogenic activities: untreated domestic sewage due to misconnections in a periurban river, effluent from urban WWTPS, agricultural runoff (nitrates) in several streams, discharge from a paper mill (humic-like substances due to wood processing) and from steel mills (PAHs).

Activated sludge pilot plant: comparison between experimental and predicted concentration profiles using three different modelling approaches.

This paper presents an experimental and numerical study of an activated sludge channel pilot plant. Concentration profiles of oxygen, COD, NO(3) and NH(4) have been measured for several operating conditions. These profiles have been compared to the simulated ones with three different modelling approaches, namely a systemic approach, CFD and compartmental modelling. For these three approaches, the kinetics model was the ASM-1 model (Henze et al., 2001). The three approaches allowed a reasonable simulation of all the concentration profiles except for ammonium for which the simulations results were far from the experimental ones. The analysis of the results showed that the role of the kinetics model is of primary importance for the prediction of activated sludge reactors performance. The fact that existing kinetics parameters in the literature have been determined by parametric optimisation using a systemic model limits the reliability of the prediction of local concentrations and of the local design of activated sludge reactors.

Arsenic mass balance in a paper mill and impact of the arsenic release from the WWTP effluent on the Moselle River.

Rivers used for drinking water production might be subject to anthropogenic pollution discharge upstream of the intake point. This problem was investigated in the case of the Moselle River, used for water production in Nancy (350,000 inhabitants) and which might be impacted by industrial activities 60 km upstream. The arsenic flux of a pulp and paper mill discharging in the Moselle River at this location has been more specifically investigated. The main sources of arsenic in that mill seemed to be the recovered papers and the gravel pit water used as feed water. The arsenic input related to wood and bark was limited. The main arsenic outputs from the plant were the paper produced on site and the deinking sludge. The arsenic concentration in the effluent of the wastewater treatment plant (WWTP) was not correlated to the one in the gravel pit water, but may depend on the operating conditions of the WWTP or the changes in processes of the mill. The impact of this anthropogenic source of arsenic on the Moselle River was slightly larger in summer, when the flowrate was lower. Globally the impact of the paper mill on the Moselle River water quality was limited in terms of arsenic.

Adverse effects of erythromycin on the structure and chemistry of activated sludge.

This study examines the effects of erythromycin on activated sludge from two French urban wastewater treatment plants (WWTPs). Wastewater spiked with 10 mg/L erythromycin inhibited the specific evolution rate of chemical oxygen demand (COD) by 79% (standard deviation 34%) and the specific N-NH4+ evolution rate by 41% (standard deviation 25%). A temporary increase in COD and tryptophan-like fluorescence, as well as a decrease in suspended solids, were observed in reactors with wastewater containing erythromycin. The destruction of activated sludge flocs was monitored by automated image analysis. The effect of erythromycin on nitrification was variable depending on the sludge origin. Erythromycin inhibited the specific nitrification rate in sludge from one WWTP, but increased the nitrification rate at the other facility.

Acute sensitivity of activated sludge bacteria to erythromycin.

The presence of antibiotics in water resources has been disturbing news for the stakeholders who are responsible for public health and the drinking water supply. In many cases, biological wastewater treatment plants are the final opportunity in the water cycle to trap these substances. The sensitivity of activated sludge bacteria to erythromycin, a macrolide widely used in human medicine was investigated in batch toxicity tests using a concentration range of 1-300 mg L(-1). Erythromycin, a protein synthesis inhibitor, has been found to significantly inhibit ammonification, nitritation and nitratation at concentrations higher than 20 mg L(-1). The degree of inhibition increased with greater concentrations of the antibiotic. Exposure to erythromycin also clearly affected heterotrophs, particularly filamentous bacteria, causing floc disintegration and breakage of filaments. Cell lysis was observed with the concomitant release of organic nitrogen (intracellular proteins) and soluble COD. Although erythromycin exhibits properties of a surfactant, this characteristic alone cannot explain the damage to heterotrophs: the effects from erythromycin were greater than those of Tween 80, a commonly used surfactant. Floc disruption can lead to the release of isolated bacteria, and possibly antibiotic resistance genes, into the environment.

Wastewater fingerprinting by UV-visible and synchronous fluorescence spectroscopy.

Municipal wastewater samples have been collected in three different types of community and fingerprinted by optical methods combining UV-visible spectrometry, synchronous fluorescence spectrometry and turbidity. Correlations, whose slope depends on the sampling location, were obtained between absorbance at 254 nm and the synchronous fluorescence intensity of peaks P1 (I366/316), P2 (I430/380) and P3 (I520/470). The corresponding correlation coefficients are larger than 0.75. Although related to urine as ammonia, the fluorescence intensity of P1 does not exhibit a strong correlation with this substance (correlation coefficient of approximately 0.6). All the measured parameters exhibit diurnal variation patterns related to human activities.