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Characterization and Performance of Algal Biofilms for Wastewater Treatment and Industrial Applications Utah State University DigitalCommons@USU All Graduate Theses and Dissertations Graduate Studies 8-2015 Characterization and Performance of Algal Biofilms for Wastewater Treatment and Industrial Applications Maureen Kesaano Utah State University Follow this and additional works at: https://digitalcommons.usu.edu/etd Part of the Biological Engineering Commons Recommended Citation Kesaano, Maureen, "Characterization and Performance of Algal Biofilms for astewaterW Treatment and Industrial Applications" (2015). All Graduate Theses and Dissertations. 4445. https://digitalcommons.usu.edu/etd/4445 This Dissertation is brought to you for free and open access by the Graduate Studies at DigitalCommons@USU. It has been accepted for inclusion in All Graduate Theses and Dissertations by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. CHARACTERIZATION AND PERFORMANCE OF ALGAL BIOFILMS FOR WASTEWATER TREATMENT AND INDUSTRIAL APPLICATIONS by Maureen Kesaano A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biological Engineering Approved: _______________________ _____________________ Dr. Ronald C. Sims Dr. Byard Wood Major Professor Committee Member _______________________ ______________________ Dr. Charles D. Miller Dr. Jon Takemoto Committee Member Committee Member ________________________ ______________________ Mr. Issa Hamud Dr. Mark McLellan Committee Member Vice President for Research and Dean of the School of Graduate Studies UTAH STATE UNIVERSITY Logan, Utah 2015 ii Copyright © Maureen Kesaano 2015 All rights reserved iii ABSTRACT Characterization and Performance of Algal Biofilms for Wastewater Treatment and Industrial Applications by Maureen Kesaano, Doctor of Philosophy Utah State University, 2015 Major Professor: Dr. Ronald C. Sims Department: Biological Engineering Fundamental studies on algal biofilms grown for biological wastewater treatment and as a feedstock for biofuels production were carried out to determine: nutrient uptake capacity, biofilm growth rates, photosynthetic activity, lipid accumulation from nitrogen stress and bicarbonate addition, and algae-bacteria interactions. The algal biofilms were cultured on rotating algal biofilm reactors (RABRs) under varying conditions of light, nutrients, and species composition at bench and pilot scale. Oxygen microsensor-based measurements and microscopy were used to characterize spatial photosynthetic zones and biofilm structure, respectively. Nutrients were analyzed using ion chromatography and TOC/TN analyzer. Lipid composition and quantification were determined using gas chromatography flame ionization detection (GC-FID) and mass spectrophotometry (GC-MS). Drip flow reactors (modified to allow for phototrophic growth) were utilized to study algae-bacteria interactions under sterile conditions. iv The results showed that photosynthetic and respiration rates observed in the algal biofilms were influenced by biofilm composition, culturing conditions, orientation to the light, and nutrient availability. Nutrient removal capacity was highest during the exponential growth phase of the biofilm. A slight increase in lipid production was observed as a result of nitrogen stress and a combination of nitrogen stress with bicarbonate addition. Presence of bacteria positively influenced microalgae growth in the mixed cultures but the reverse was not true. In conclusion, spatial photosynthetic activity and biofilm structure were successfully characterized using the methods developed in this study. Algal biofilms can potentially be utilized for nutrient uptake in wastewater but their biofuels potential will depend on the amount of biomass produced because of the low lipid yields realized. Further research on algae-bacteria interactions using species native to the wastewater grown algal biofilms is recommended. (187 pages) v PUBLIC ABSTRACT Characterization and Performance of Algal Biofilms for Wastewater Treatment and Industrial Applications Maureen Kesaano This study was carried out on algal biofilms grown using rotating algal biofilm reactors (RABRs) with the aim of: i) characterizing their growth in terms of photosynthetic activity and morphology ii) evaluating their performance as a wastewater treatment option and a feedstock for biofuels production, and iii) examining the algal- bacteria interactions. A review of algal biofilm technologies currently employed in wastewater treatment processes was made to compare nutrient removal efficiencies, factors that influenced algal biofilm growth, and the different bioproducts generated from algal biomass. Consequently, research efforts were directed towards addressing pertinent issues identified in literature in order to optimize these systems for wastewater treatment and bioproducts production. Successful growth of algal biofilms in municipal wastewater and subsequent removal of nutrients from the wastewater was demonstrated. Photosynthetic and respiration rates observed with depth of the biofilm were influenced by the biofilm composition (single vs. mixed species), culturing conditions (laboratory vs. outdoor), orientation to the light, nitrogen availability (N-replete vs. N-deplete), and dissolved inorganic carbon availability (presence or absence of bicarbonate). Slight enhancement in lipid production was also observed as a result of nitrogen stress and bicarbonate addition. vi However, the accumulated lipids were not as much as expected or as reported in suspended cultures. Presence of bacteria positively influenced microalgae growth in the mixed cultures but the reverse was not true. In conclusion, photosynthetic activity and biofilm structure were characterized with methods developed for the algal biofilms in this study. For now, productivity of the algal biofilms needs to be maximized in order to fully utilize its potential as a biofuel feedstock and nutrient removal option. Further research on algae-bacteria interactions using species native to the wastewater grown algal biofilms is recommended. vii DEDICATIONS This dissertation is dedicated to my mom, Dr. Margaret Saimo-Kahwa. I am indebted to your support and encouragement through this journey. viii ACKNOWLEDGMENTS I would like to thank my advisor, Dr. Ronald C. Sims, whose support and advice has been invaluable; my committee members, Dr. Charles Miller, Mr. Issa Hamud, Dr. Jon Takemoto, and Dr. Byard Wood, for their assistance throughout this process; my family and friends for their patience and unwavering support; and the almighty God for life and good health. Special thanks to the Utah State University Sustainable Waste to Bioproducts (SWBEC) algal research group, the staff at Montana State University Center for Biofilm Engineering (CBE), especially Ms. Betsey Pitts and Anne Willis; Montana State University algal research group members, Katie Davis, Luke Halverson, Tisza Bell, and Todd Pedersen; my co-authors, Karen Moll, Bethany Jensen, Ryan Putman, Drs. Robert Gardner, Hans Bernstein, Ellen Lauchnor, Keith Cooksey, Ross Carlson, Robin Gerlach, and Brent Peyton. I am also grateful for the funding support from: (1) the Logan City Environmental Department, (2) the Utah Water Research Laboratory, (3) the Huntsman Environmental Research Center, (4) the Sustainable Waste to Bioproducts Engineering Center, and (5) the Utah Science Technology and Research Initiative (USTAR) program. Maureen Kesaano ix CONTENTS Page ABSTRACT……………………………………………………………………...………iii PUBLIC ABSTRACT…………………………………………………………………….v DEDICTATIONS………………………………………………………………………..vii ACKNOWLEDGMENTS……………………………………………………………....viii LIST OF TABLES………………………………………………………..………….....xiii LIST OF FIGURES………………………………………………………………...…...xiv CHAPTER 1. INTRODUCTION…..……..……..…………………………………………...1 Introduction………………………………..…………………………………..1 Photosynthesis and photorespiration in microalgae cultures……..…..……….3 Influence of bicarbonate on algal biofilm growth, nutrient uptake, and lipid accumulation……………………………………..……………………..4 Algae-bacteria interactions in mixed cultures………..………………………..6 General overview and summary of the dissertation…………..……………….6 Justification of the study…………………………..…………………………..7 Refereed publications………………………………………………………….7 References……………..………………………………………………………8 2. ALGAL BIOFILM-BASED TECHNOLOGY FOR WASTEWATER TREATMENT- A REVIEW…………………………………………………12 Abstract……………………………………………..………………………..12 Introduction……………………..……………………………………...…….13 Attached growth/fixed film technologies in wastewater treatment……...…..14 Algal biofilm-based nutrient removal option………………………..……….15 Generation of algal biomass for bioproducts production...……………..……19 Characterization of algal biofilm growth conditions.………………..…...….23 Light….…………………………………………………………………..24 Temperature…………………………………………………..………….26 Nutrients……………………………………...…………………………..28 Substratum………………………………………….……………………30 Extracellular polymeric substances (EPS)……………………….………32 x Species interaction…………………………………….…………………33 Interaction of environmental and biological factors…………………………34 Recommendations…………..………………………………………………..35 Conclusions………………………………………..…………………………36 References…………………………………………..………………………..36 3. DISSOLVED INORGANIC CARBON-ENHANCED GROWTH, NUTRIENT UPTAKE, AND LIPID ACCUMULATION IN WASTEWATER-GROWN MICROALGAL BIOFILMS……………..……47 Abstract……..…………………………………………..……………………47 Introduction……………………………..……………………...…………….48 Materials and methods…………………………………..………….………..50 Microalgal biofilm culturing and sampling……………………………...50 Water quality monitoring…….………………………………………..…52
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