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Controlling Anaerobic Digestion to Produce Targeted Compounds

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Controlling Anaerobic Digestion to Produce Targeted Compounds Controlling anaerobic digestion to produce targeted compounds Miriam Peces Gomez Bachelor in Chemical Engineering Master in Environmental Engineering A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2017 School of Civil Engineering Centre for Solid Waste Bioprocessing Abstract Anaerobic digestion is a mix-culture microbial-mediated process that has primarily been applied to produce methane and stabilise organic matter. However, the intermediates of the digestion process (volatile fatty acids, alcohols, and hydrogen) have applications as commodity chemicals or as precursors to a range of biobased products. However, one of the main challenges to broaden the application of anaerobic digestion is the difficulty associated with robustly controlling mixed-culture products, such that a suite of products can be repeatedly produced. Therefore, understanding how the change in microbial populations or loss in microbial functionality influence the behaviour of the rest of the community can prove to be a powerful tool for manipulating and controlling processes towards a desired commodity. The impact of the starting inoculum on long-term anaerobic digestion performance, metabolic activity rates and microbial community composition remains unclear. To understand the impact of starting inoculum, active microbial communities from four different full-scale anaerobic digesters were each used to inoculate four continuous anaerobic digesters. Thereafter, the digesters were -1 -1 operated identically at 15 days solid retention time, an organic loading rate of 1 g COD Lr d (75:25 - cellulose:casein), and 37 ºC for 295 days. The digesters performance converged and stabilised in 80 days, while activity rates and microbial communities converged and stabilised after 145 days of operation. After 295 days, 52% of all identified OTUs were common to all digesters, and this core community accounted for 72% of the total microbial community relative abundance defined by various bacterial taxa (Bacteroidales, Ruminoccocaceae, Kosmotoga and Treponema) and archaeal taxa (Methanosaeta, Candidatus Methanoregula and Methanospirillum). This indicates that deterministic factors (process operational conditions) were a stronger driver in controlling the ultimate microbial composition in a digester rather than the initial microbial community composition. Moreover, Pearson correlation coefficients revealed several significant associations between bacterial taxa found in the digesters and activity rate profiles. For instance, the presence of Armatimonadetes was positively correlated with higher cellulolytic rates and bacteria belonging to genus Synthophobacter and Clostridum or families Veillonellaceae and candidate BA008 (phylum Bacteroidetes) were correlated to higher butyrate and propionate degradation rates. Overall, it seems plausible that process operational conditions can be used to tune microbial composition and functionality in an anaerobic digester. To explore the extent that the anaerobic digestion process can be manipulated by a sole selection pressure, the solid retention time was isolated as a pressure parameter. Without interruption, the same four continuous anaerobic digesters were subjected to a sequential decrease in solid retention time -1 -1 from 15 to 8 to 4 to 2 days while maintaining the organic loading rate at 1 g COD Lr d , the same i substrate composition ratio (75:25 - cellulose:casein) and the same temperature (37 ºC). Each solid retention time was operated until steady state was achieved. Results showed that acetoclastic methanogenesis carried out by Methanosaeta remained active down to 2 day solid retention time and only minor accumulation of volatile fatty acids was achieved (less than 3.5% of influent COD). Therefore, solid retention time as an individual selection pressure was not an effective parameter to shift the anaerobic digestion product profile. However, lowering solid retention times induced a shift in metabolic activity rates, where ethanol degradation gained dominance over butyrate and propionate degradation. Solid retention time also influenced the microbial dynamics of the digesters, driving changes at family or genus level, although the most noticeable finding was the formation of biofilms containing a high abundance of Methanosaeta at the lowest solid retention time. This suggests that the different microbial communities in all four digesters developed similar survival strategies under non-favourable methanogenic conditions. To contextualise and prove the applicability of imposed conditions to steer the process, a combination of temperature, retention time and oxygen availability were selected to control the fermentation patterns of primary sludge followed by anaerobic digestion to recover biogas, as part of a bio-refinery concept. Primary sludge pre-fermentation was carried out at different temperatures (20, 37, 55, 70ºC), treatment times (12, 24, 48, 72h), and oxygen availability (semi-aerobic, anaerobic). pH was not controlled. The anaerobic biodegradability after pre-fermentation was evaluated using biochemical methane potential tests. The results showed that fermentation at 20 and 37 ºC was optimal for volatile fatty acids production with acetate and propionate being major products. Anaerobic fermentation at 37, 55 and 70 ºC resulted in higher solubilisation yield at the expense of reduced methane production by 20%, while semi-aerobic fermentation allowed both volatile fatty acids recovery and improved methane potential. Replication experiments using a different batch of primary sludge showed that the main trends could be reproduced exemplifying that fermentation and anaerobic digestion products can be controlled by operational decisions. ii Declaration by author This thesis is composed of my original work, and contains no material previously published or written by another person except where due reference has been made in the text. I have clearly stated the contribution by others to jointly-authored works that I have included in my thesis. I have clearly stated the contribution of others to my thesis as a whole, including statistical assistance, survey design, data analysis, significant technical procedures, professional editorial advice, and any other original research work used or reported in my thesis. The content of my thesis is the result of work I have carried out since the commencement of my research higher degree candidature and does not include a substantial part of work that has been submitted to qualify for the award of any other degree or diploma in any university or other tertiary institution. I have clearly stated which parts of my thesis, if any, have been submitted to qualify for another award. I acknowledge that an electronic copy of my thesis must be lodged with the University Library and, subject to the policy and procedures of The University of Queensland, the thesis be made available for research and study in accordance with the Copyright Act 1968 unless a period of embargo has been approved by the Dean of the Graduate School. I acknowledge that copyright of all material contained in my thesis resides with the copyright holder(s) of that material. Where appropriate I have obtained copyright permission from the copyright holder to reproduce material in this thesis. iii Publications during candidature Peer-reviewed journal papers Puyol, D., Batstone, D.J., Hülsen, T., Astals, S., Peces, M., Krömer, J.O. 2017. Resource recovery from wastewater by biological technologies: Opportunities, challenges, and prospects. Frontiers in Microbiology, 7, article 2106. Peces, M., Astals, S., Clarke, W.P. and Jensen, P.D., 2016. Semi-aerobic fermentation as a novel pre-treatment to obtain VFA and increase methane yield from primary sludge. Bioresource technology, 200, pp.631-638. Nolla-Ardèvol, V., Peces, M., Strous, M., Tegetmeyer, H.E., 2015. Metagenome from a Spirulina digesting biogas reactor: Analysis via binning of contigs and classification of short reads. BMC Microbiology, 15-1, article 15. Peces, M., Astals, S., Mata-Alvarez, J., 2015. Effect of moisture on pretreatment efficiency for anaerobic digestion of lignocellulosic substrates. Waste Management, 46, pp.189-196. Conference papers Peces, M., Jensen, P.D., Astals, S., and Clarke, W.P. 2016. Do different inocula converge given the same operational conditions in long-term anaerobic digestion? In proceedings: XII DAAL - Taller y Simposio Latino Americano en Digestión Anaerobia. Cusco, Peru. (Conference oral presentation) Peces, M., Jensen P.D., Clarke, W.P. and Astals, S., 2015. Semi-aerobic pre-fermentation conditions to recover VFA and improve methane potential. In proceedings: 14th World Congress on Anaerobic Digestion. Viña del Mar, Chile. (Conference oral presentation) iv Publications included in this thesis Peces, M., Astals, S., Clarke, W.P. and Jensen, P.D., 2016. Semi-aerobic fermentation as a novel pre-treatment to obtain VFA and increase methane yield from primary sludge. Bioresource technology, 200, pp.631-638. This paper has been modified an incorporated as Chapter 7. Contributor Statement of contribution Miriam Peces (Candidate) Designed experiments (30%) Conducted experiments (80%) Data analysis (90%) Wrote the paper (60%) Sergi Astals Designed experiments (70%) Conducted experiments (20%) Data analysis (10%) Wrote the paper (30%) Critically reviewed and edited the paper (40%) William P. Clarke Wrote paper (5%) Critically reviewed and edited the paper (20%) Paul D. Jensen Wrote paper (5%) Critically
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