• Energy Research

The valorization of glycerol wastes through its conversion into high-added value products using biotechnological processes is very promising due to the high amount of glycerol generated from the bio-diesel industry. In this sense, it is important to develop rapid, reliable and, if possible, environmentally friendly analytical techniques for the monitoring of high-added value compounds. With this in mind, Fourier transform near infrared (FT-NIR) spectroscopy was explored for the assessment of biomass, glycerol, l-phenylalanine and 2-phenylethanol (2-PE) parameters during the biotechnological production of 2-PE. A total of 80 samples were collected and analyzed through FT-NIR spectroscopy. Partial least squares (PLS) regression method was selected for establishing quantification models. The obtained figures of merit, in terms of, coefficients of determination of prediction (R2P), range error-ratios (RER) and residual predictive deviation (RPD) yielded values higher than 0.85, 7.7 and 2.6, respectively, for all the parameters assessed. The obtained results reveal that all the developed PLS models can be considered as models with a good predictive accuracy and therefore attest the suitability of FT-NIR spectroscopy for the quantification of these parameters. Concluding, a new rapid, reliable, cost-effective, accurate, multi-parametric and environmentally friendly analytical technique exploring the potential of FT-NIR spectroscopy was successfully developed. The authors thank the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic fundings of UIDB/ BIO/04469/2020 and UIDB/QUI/50006/2020 units. Ricardo N.M.J. Páscoa and Cristina Quintelas thanks FCT for funding through program DL 57/2016 – Norma transitória. The authors also want to express their gratitude to Prio Energy – Prio Biocombustíveis, SA for providing the glycerol used in this experimental work. info:eu-repo/semantics/publishedVersion

Bacterial cellulose (BC), obtained by fermentation, is an innovative and promising material with a broad spectrum of potential applications. Despite the increasing efforts towards its industrialization, a deeper understanding of the environmental impact related to the BC production process is still required. This work aimed at quantifying the environmental, health, and resource depletion impacts related to a production of BC. An attributional life cycle assessment (LCA) was applied to a process design of production of BC, by static culture, following a cradle-to-gate approach. The LCA was modeled with GaBi Pro Software using the ReCiPe 2016 (H) methodology with environmental impact indicators at midpoint level. The functional unit was defined as 1 kg of BC (dry mass), in 138.8 kg of water. From the total used resources (38.9 ton/kg of BC), water is the main one (36.1 ton/kg of BC), most of which (98%) is returned to fresh waters after treatment. The production of raw materials consumed 17.8 ton of water/kg of BC, 13.8 ton/kg of BC of which was for the production of carton packaging, culture medium raw materials, and sodium hydroxide (for the washing of BC). The remaining consumed water was mainly for the fermentation (3.9 ton/kg) and downstream process (7.7 ton/kg). From the identified potential environmental impacts, the production of raw materials had the highest impact, mainly on “Climate change”, “Fossil depletion”, “Human toxicity, non-cancer”, and “Terrestrial toxicity”. The sodium dihydrogen phosphate production, used in the culture medium, showed the highest environmental impacts in “Human toxicity, non-cancer” and “Terrestrial ecotoxicity”, followed by corn syrup and carton production. The static culture fermentation and downstream process showed impact in “Climate change” and “Fossil depletion”. Per se, the BC production process had a small contribution to the consumption of resources and environmental impact of the BC global life cycle.

An aerobic granular bioreactor was operated for over 4months, treating a synthetic wastewater with a high ammonium content (100mgNL-1). The inoculum was collected from a bioreactor performing simultaneous partial nitrification and aromatic compounds biodegradation. From day-56 onwards, 2-fluorophenol (2-FP) (12.4mgL-1) was added to the feeding wastewater and the system was bioaugmented with a 2-FP degrading bacteria (Rhodococcus sp. FP1). By the end of operation, complete 2-FP biodegradation and partial nitrification were simultaneously achieved. Aerobic granules remained stable over time. During the 2-FP loading, a shift in the community structure occurred, coinciding with the improvement of 2-FP degradation. DGGE analysis did not allow to infer on the bioaugmented strain presence but pyrosequencing analysis detected Rhodococcus genus by the end of operation. Together with other potential phenolic-degraders within granules, these microorganisms were probably responsible for 2-FP degradation.

In the last years, Salmonella has been extensively studied not only due to its importance as a pathogen, but also as a host to produce pharmaceutical compounds. However, the full exploitation of Salmonella as a platform for bioproduct delivery has been hampered by the lack of information about its metabolism. Genome-scale metabolic models can be valuable tools to delineate metabolic engineering strategies as long as they closely represent the actual metabolism of the target organism. In the present study, a 13C-MFA approach was applied to map the fluxes at the central carbon pathways of S. typhimurium LT2 growing at glucose-limited chemostat cultures. The experiments were carried out in a 2L bioreactor, using defined medium enriched with 20% 13C-labeled glucose. Metabolic flux distributions in central carbon pathways of S. typhimurium LT2 were estimated using OpenFLUX2 based on the labeling pattern of biomass protein hydrolysates together with biomass composition. The results suggested that pentose phosphate is used to catabolize glucose, with minor fluxes through glycolysis. In silico simulations, using Optflux and pFBA as simulation method, allowed to study the performance of the genome-scale metabolic model. In general, the accuracy of in silico simulations was improved by the superimposition of estimated intracellular fluxes to the existing genome-scale metabolic model, showing a better fitting to the experimental extracellular fluxes, whereas the intracellular fluxes of pentose phosphate and anaplerotic reactions were poorly described.

The use of quantitative image analysis techniques, together with physiological information might be used to monitor and detect operational problems in advance to reactor performance failure. Industrial organic solvents, such as White Spirit, are potentially harmful to granular sludge. In preliminary batch assays, 33 mg L(-1) of solvent caused 50% relative biomass activity loss. In an expanded granular sludge blanket reactor fed with 40 mg L(-1) of solvent, during 222h, the reactor performance seemed to be unaffected, presenting COD removal efficiency consistently >95%. However, in the last days of exposure, the biogas production and the methane content were inhibited. Afterwards, already during recovery phase, the COD removal efficiency decreased to 33%, probably because the reactor was underloaded and the biomass became saturated in solvent only at this stage. In the first hours of exposure the specific acetoclastic and the specific hydrogenotrophic methanogenic activities decreased 29% and 21%, respectively. The % of aggregates projected area with equivalent diameter (D(eq))>1mm decreased from 81% to 53%. The mean D(eq) of the aggregates > or =0.2mm decreased, as well as the settling velocity, showing that the granules experienced fragmentation phenomenon caused by the solvent shock load. The ratio between total filaments length and total aggregates projected area (LfA) increased 2 days before effluent volatile suspended solids, suggesting that LfA could be an early-warning indicator of washout events.

Fishery wastewater treatment can be compromised due to seasonal production. The use of sequencing batch reactors is not completely successful, despite flexibility being one of the principal advantages. Most research on activated sludge is performed using synthetic wastewater to ensure a stable and constant feed. The current work compared biomass morphology and settling ability using image analysis of synthetic and real fishery wastewaters, with and without NaCl addition.The final effluent presented higher turbidity for both wastewaters after NaCl addition, and lower SVI values. For synthetic wastewater, NaCl addition led to the total aggregates' area (TA) increase from 1.46 to 2.09 mm(2)/μL, alongside the growth of intermediate aggregates into larger aggregates. The addition of NaCl to the fishery wastewater led to a decrease of the TA from 4.43 to 1.72 mm(2)/μL. The biomass composition decreased in larger and intermediate structures, opposite to the smaller aggregates' area percentage increase.NaCl addition to synthetic wastewater incited flocculation increasing sludge settling ability. A slight aggregate disruption was responsible for a turbidity increase. A strong deflocculation was identified in fishery wastewater with NaCl from the decrease of intermediate and large aggregates. This contrasted with pinpoint flocs release, which increased the turbidity levels. CONCLUSIONS, RECOMMENDATIONS, AND PERSPECTIVES: It could be established that synthetic wastewater biomass flocculation and fishery wastewater biomass deflocculation, observed during 0.5% NaCl experiments, were related to sludge settling and effluent turbidity changes. Furthermore, the biomass changes obtained with synthetic wastewater cannot be extrapolated to fishery wastewater.

AbstractImage analysis techniques are applied to monitor the morphological changes in granular sludge present in an expanded granular sludge blanket (EGSB) reactor fed with oleic acid. Deterioration of granular sludge was monitored along the trial period by measuring the percentage of aggregates smaller than 1 mm (in terms of Feret diameter) either in terms of projected area or in terms of number of aggregates. A good correlation was obtained between these values and the percentage of aggregates smaller than 1 mm were physically sorted and quantified by the volatile suspended solid content. The ratio of total filaments length to cross‐sectional area of aggregates defined as LfA, was applied to quantify the dispersion level of the granular sludge, which increased until day 141 and remained almost invariant afterwards. LfA was sensitive to the sludge deterioration process and was able to indicate, with the anticipation of about 1 month, the most significant biomass washout episode that occurred in the trial period. A mechanism of filaments' release, detachment and selective washout was proposed to explain the action of LfA from this viewpoint. The equivalent diameter of the bottom aggregates larger than 1 mm increased with the increase on the amount of long chain fatty acids associated with the biomass by mechanisms of adsorption, precipitation, or entrapment. After a threshold value of about 200 mg COD‐LCFA gVSS (COD = chemical oxygen demand; LCFA = long chain fatty acids; VSS = volatile suspended solids), a migration of granular sludge from the bottom to a top‐floating layer was evident. © 2004 Wiley Periodicals, Inc.