In this study, a 1200L outdoor pilot scale microalgal photobioreactor (PBR) was used for toilet wastewater (WW) treatment and evaluate its ability to remove pharmaceutically active compounds (PhACs). The PBR was operated at two different hydraulic retention times (HRTs), which were 8 and 12days, during Period I (September-October) and Period II (October-December), respectively. Algal biomass concentrations varied by operating period because of seasonal changes. Nutrients (ammonia, nitrogen and total phosphorous) and chemical oxygen demand (COD) were monitored and efficiently removed in both periods (>80%), attaining the legislation limits. At the theoretical hydraulic steady state in both periods, pharmaceutical removal reached high levels (>48%). Two harvesting techniques were applied to the PBR microalgae effluent. Gravity sedimentation was efficient for biomass removal (>99% in 7min) in Period I when large particles, flocs and aggregates were present. In contrast, a longer sedimentation time was required when biomass was mainly composed of single cells (88% clarification in a 24h in Period II). The second harvesting technique investigated was the co-pelletization of algal biomass with the ligninolytic fungus Trametes versicolor, attaining >98% clarification for Period II biomass once pellets were formed. The novel technology of co-pelletization enabled the complete harvesting of single algae cells from the liquid medium in a sustainable way, which benefits the subsequent use of both biomass and the clarified effluent.

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Please cite this article as: Passos, F., Hom-Diaz, A., Blanquez, P., Vicent, T., Ferrer, I., Improving biogas production from microalgae by enzymatic pretreatment, Bioresource Technology (2015), doi: http://dx.doi.org/10. 1016/ j.biortech.2015.08.084 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Improving biogas production from microalgae by enzymatic pretreatment AbstractIn this study, enzymatic pretreatment of microalgal biomass was investigated under different conditions and evaluated using biochemical methane potential (BMP) tests. Cellulase, glucohydrolase and an enzyme mix composed of cellulase, glucohydrolase and xylanase were selected based on the microalgae cell wall composition (cellulose, hemicellulose, pectin and glycoprotein). All of them increased organic matter solubilisation, obtaining high values already after 6 hours of pretreatment with an enzyme dose of 1% for cellulase and the enzyme mix. BMP tests with pretreated microalgae showed a methane yield increase of 8 and 15% for cellulase and the enzyme mix, respectively. Prospective research should evaluate enzymatic pretreatments in continuous anaerobic reactors so as to estimate the energy balance and economic cost of the process.

The evaluation of wastewater treatment capabilities in terms of removal of water pollutants is crucial when assessing water mitigation issues. Not only the monitoring of target pollutants becomes a critical point, but also the transformation products (TPs) generated. Since these TPs are very often unknown compounds, their study in both wastewater and natural environment is currently recognized as a tedious task and challenging research field. In this study, a novel automated suspect screening methodology was developed for a comprehensive assessment of the TPs generated from nine antibiotics during microalgae water treatment. Three macrolides (azithromycin, erythromycin, clarithromycin), three fluoroquinolones (ofloxacin, ciprofloxacin, norfloxacin) and three additional antibiotics (trimethoprim, pipemidic acid, sulfapyridine) were selected as target pollutants. The analysis of samples was carried out by direct injection in an on-line turbulent flow liquid chromatography-high resolution mass spectrometry (TFC-LC-LTQ-Orbitrap-MS/MS) system, followed by automatic data processing for compound identification. The screening methodology allowed the identification of 40 tentative TPs from a list of software predicted intermediates created automatically. Once known and unknown TPs were identified, degradation pathways were suggested considering the different mechanisms involved on their formation (biotic and abiotic). Results reveal microalgae ability for macrolide biotransformation, but not for other antibiotics such as for fluoroquinolones. Finally, the intermediates detected were included into an in-house library and applied to the identification of tentative TPs in real toilet wastewater treated in a microalgae based photobioreactor (PBR). The overall approach allowed a comprehensive overview of the performance of microalgae water treatment in a fast and reliable manner: it represents a useful tool for the rapid screening of wide range of compounds, reducing time invested in data analysis and providing reliable structural identification.

Coupling microalgae production to wastewater treatment can reduce the costs of microalgae production for non-food bioproducts and energy consumption for wastewater treatment. Furthermore, microalgae anaerobic digestion can be enhanced by applying pretreatment techniques. The aim of this study is to improve the biogas production from microalgal biomass grown in urban wastewater treatment systems by applying an enzymatic pretreatment with crude fungal broth and commercial laccase. To this end, the fungus Trametes versicolor was cultured, and the enzymatic activity of the culture broth analysed by measuring laccase concentration. The results showed that both the fungal broth and commercial laccase pretreatment (100 U L- 1) over an exposure time of 20 min increased the methane yield in batch tests. Indeed, the fungal broth pretreatment increased the methane yield by 74%, while commercial laccase increased the methane yield by 20% as compared to non-pretreated microalgal biomass. In this manner, laccase addition enhanced microalgal biomass anaerobic biodegradability, and addition of T. versicolor broth further improved the results. This fact may be attributed to the presence of other molecules excreted by the fungus.Peer ReviewedPostprint (author's final draft

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The use of microalgal consortia for urban wastewater treatment is an increasing trend, as it allows simultaneous nutrient removal and biomass production. Emerging contaminants proposed for the list of priority substances such as the hormone 17β-estradiol are commonly found in urban wastewater, and their removal using algal monocultures has been accomplished. Due to the inherent potential of algae-based systems, this study aimed to assess the capability of native photobioreactor biomass to remove 17β-estradiol under indoor and outdoor conditions. At the same time, the microbial community changes in regular and bioaugmented operations with Scenedesmus were assessed. The results show that almost complete removal (>93.75%) of the hormone 17β-estradiol can be attained in the system under favourable seasonal conditions, although these conditions greatly influence biomass concentrations and microbial diversity. Even under the harsh conditions of low temperatures and solar irradiation, the established consortium removed more than 50% of the pollutant in 24 h. While species from genus Chlorella were stable during the entire operation, the microbial diversity analysis revealed that assorted and evenly distributed populations stimulate the removal rates. Bioaugmentation assays proved that the input of additional biomass results in higher overall removal and decreases the yield per mg of biomass.