16 2 The influence of biogas flow rate (0, 0.3, 0.6 and 1.2 m 3 m -2 h -1 ) on the elemental and 17 macromolecular composition of the algal-bacterial biomass produced from biogas upgrading in a 18 180 L photobioreactor interconnected to a 2.5 L external bubbled absorption column was 19 investigated using diluted anaerobically digested vinasse as cultivation medium. The influence of 20 the external liquid recirculation/biogas ratio (0.5 < L/G < 67) on the removal of CO2 and H2S, 21and on the concentrations of O2 and N2 in the upgraded biogas was also evaluated. A L/G ratio of 22 10 was considered optimum to support CO2 and H2S removals of 80% and 100%, respectively, at 23 all biogas flow rates tested. Biomass productivity increased at increasing biogas flow rate, with a 24 maximum of 12±1 g m -2 d -1 at 1.2 m 3 m -2 h -1 , while the C, N and P biomass content remained 25 constant at 49±2%, 9±0% and 1±0%, respectively, over the 175 days of experimentation.
The microbial population from a reactor using methane as electron donor for denitrification under microaerophilic conditions was analyzed. High numbers of aerobic methanotrophic bacteria (3 10(7) cells/ml) and high numbers of acetate-utilizing denitrifying bacteria (2 10(7) cells/ml) were detected, but only very low numbers of methanol-degrading denitrifying bacteria (4 10(4) cells/ml) were counted. Two abundant acetate-degrading denitrifiers were isolated which, based on 16S rRNA analysis, were closely related to Mesorhizobium plurifarium (98.4% sequence similarity) and a Stenotrophomonas sp. (99.1% sequence similarity). A methanol-degrading denitrifying bacterium isolated from the bioreactor morphologically resembled Hyphomicrobium sp. and was moderately related to H. vulgare (93.5% sequence similarity). The initial characterization of the most abundant methanotrophic bacterium indicated that it belongs to class II of the methanotrophs. "In vivo" 13C-NMR with concentrated cell suspensions showed that this methanotroph produced acetate under oxygen limitation. The microbial composition of reactor material together with the NMR experiments suggest that in the reactor methanotrophs excrete acetate, which serves as the direct electron donor for denitrification.
A wide range of pharmaceuticals and personal care products (PPCPs) are present in the environment, and many of their adverse effects are unknown. The environmental risk assessment of 26 PPCPs of relevant consumption and occurrence in the aquatic environment in Spain was accomplished in this research. Based on the ecotoxicity values obtained by bioluminescence and respirometry assays and by predictions using the US EPA ecological structure-activity relationship (ECOSAR™), the compounds were classified following the Globally Harmonized System of Classification and Labelling of Chemicals. According to the criteria of the European Medicines Agency, the real risk of impact of these compounds in wastewater treatment plants (WWTPs) and in the aquatic environment was predicted. In at least two ecotoxicity tests, 65.4 % of the PPCPs under study showed high toxicity or were harmful to aquatic organisms. The global order of the species' sensitivity to the PPCPs considered was as follows: Vibrio fischeri (5 min) > Vibrio fischeri (15 min) > algae > crustaceans > fish > biomass of WWTP. Acetaminophen, ciprofloxacin, clarithromycin, clofibrate, ibuprofen, omeprazole, triclosan, parabens and 1,4-benzoquinone showed some type of risk for the aquatic environments and/or for the activated sludge of WWTPs. Development of acute and chronic ecotoxicity data, the determination of predicted and measured environmental concentrations of PPCPs, the inclusion of metabolites and transformation products and the evaluation of mixtures of these compounds will allow further improvements of the results of the ERAs and, finally, to efficiently identify the compounds that could affect the environment.
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