Effect of ibuprofen on extracellular polymeric substances (EPS) production and composition, and assessment of microbial structure by quantitative image analysis.

A Sequencing Batch Reactor (SBR) with activated sludge was operated with synthetic wastewater containing ibuprofen (IBU) to investigate the biomass stress-responses under long-term IBU exposure. There were 3 different phases: phase I as the control without IBU for 56 days, phase II (40 days), and phase III (60 days) containing IBU at 10 and 5 mg L-1 each. The overall performance of the SBR as well as the extracellular polymeric substances (EPS) in terms of polysaccharides, proteins, and humic acid substances were estimated. Morphological parameters of microbial aggregates in the presence of IBU (phase II and phase III) were assessed by quantitative image analysis (QIA). Removal efficiencies of chemical oxygen demand (COD) and ammonium (NH4+) were significantly reduced by IBU. Loosely bound EPS (LB-EPS) decreased during phase II and phase III, and tightly bound EPS (TB-EPS) was slightly higher in phase II than phase I. TB-EPS proteins were greater in phase II, perhaps to protect microbial cells from IBU exposure. These findings provided insight into both activated sludge stress-responses and EPS composition under long-term IBU exposure. Spearman correlation showed that EPS and morphological parameters significantly affected sludge settleability and flocculation. QIA also proved to be a powerful technique in investigating dysfunctions in activated sludge under IBU exposure.

Assessment of an aerobic granular sludge system in the presence of pharmaceutically active compounds by quantitative image analysis and chemometric techniques.

In this study, a sequencing batch reactor (SBR) with aerobic granular sludge (AGS) was operated with synthetic wastewater containing environmental relevant concentrations of 17ß-estradiol (E2), 17α-ethinylestradiol (EE2) and sulfamethoxazole (SMX). Despite the presence of the studied PhAC, the granular fraction clearly predominated (TSSgran/TSS ranging from 0.82 to 0.98) throughout the monitoring period, presenting aggregates with high organic fraction (VSS/TSS above 0.83) and good settling characteristics (SVI5 ranging from 15 to 39 mL/gTSS). A principal component analysis (PCA) with quantitative image analysis (QIA) based data allowed to distinguish the different operational periods, namely with mature granules (CONT), and the E2, EE2, and SMX feeding periods. It further revealed a positive relationship between the biomass density, sludge settling ability, overall and granular biomass contents, granulation properties, granular biomass fraction and large granules fraction and size. Moreover, a discriminant analysis (DA) allowed to successfully discriminate not only the different operational periods, mainly by using the floccular apparent density, granular stratification and contents data, but also the PhAC presence in samples. The filamentous bacteria contents, sludge settling properties, settling properties stability and granular stratification, structure and contents parameters were found to be crucial for that purpose.

Economic analysis and environmental impact assessment of three different fermentation processes for fructooligosaccharides production.

Three different fermentation processes for the production of fructooligosaccharides (FOS) were evaluated and compared in terms of economic aspects and environmental impact. The processes included: submerged fermentation of sucrose solution by Aspergillus japonicus using free cells or using the cells immobilized in corn cobs, and solid-state fermentation (SSF) using coffee silverskin as support material and nutrient source. The scale-up was designed using data obtained at laboratory scale and considering an annual productivity goal of 200 t. SSF was the most attractive process in both economic and environmental aspects since it is able to generate FOS with higher annual productivity (232.6 t) and purity (98.6%) than the other processes; reaches the highest annual profit (6.55 M€); presents the lowest payback time (2.27 years); and is more favourable environmentally causing a lower carbon footprint (0.728 kg/kg, expressed in mass of CO2 equivalent per mass of FOS) and the lowest wastewater generation.

Automatic identification of activated sludge disturbances and assessment of operational parameters.

Activated sludge systems are prone to be affected by changes in operating conditions leading to problems such as pinpoint flocs formation, filamentous bulking, dispersed growth, and viscous bulking. These problems are often related with the floc structure and filamentous bacteria contents. In this work, a lab-scale activated sludge system was operated sequentially obtaining filamentous bulking, pinpoint floc formation, viscous bulking and normal conditions. Image processing and analysis techniques were used to characterize the contents and structure of aggregated biomass and the contents of filamentous bacteria. Further principal component and decision trees analyses permitted the identification of different conditions from the collected morphological data. Furthermore, a partial least squares analysis allowed to estimate the sludge volume index and suspended solids key parameters. The obtained results show the potential of image analysis procedures, associated with chemometric techniques, in activated sludge systems monitoring.

Assessment of physiological conditions in E. coli fermentations by epifluorescent microscopy and image analysis.

The development of monitoring methods for assessing the physiological state of microorganisms during recombinant fermentation processes has been encouraged by the need to evaluate the influence of processing conditions in recombinant protein production. In this work, a technique based on microscopy and image analysis was developed that allows the simultaneous quantification of parameters associated with viability and fluorescent protein production in recombinant Escherichia coli fermentations. Images obtained from light microscopy with phase contrast are used to assess the total number of cells in a given sample and, from epifluorescence microscopy, both protein producing and injured cells are evaluated using two different fluorochromes: propidium iodide and enhanced yellow fluorescent protein. This technique revealed the existence of different cell populations in the recombinant E. coli fermentation broth that were evaluated along four batch fermentations, complementing information obtained with standard techniques to study the effects of the temperature and induction time in recombinant protein production processes.