Handbook of microalgal culture applied phycology and biotechnology edited by amos richmond, qiang hu

Handbook of Microalgal Culture Applied Phycology and Biotechnology Second Edition Edited by Amos Richmond, Ph.D., Prof Emeritus Ben Gurion University of the Negev at Sede-Boker, Israel The Blaustien Institutes for Desert Research Qiang Hu, Ph.D Professor, Laboratory for Algae Research and Biotechnology Co-Director, Arizona Center for Algae Technology and Innovation, Arizona State University, Arizona This edition first published 2013 C 2004, 2013 by John Wiley & Sons, Ltd Wiley-Blackwell is an imprint of John Wiley & Sons, formed by the merger of Wiley’s global Scientific, Technical and Medical business with Blackwell Publishing Registered office: John Wiley & Sons, Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK Editorial offices: 9600 Garsington Road, Oxford, OX4 2DQ, UK The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK 111 River Street, Hoboken, NJ 07030-5774, USA For details of our global editorial offices, for customer services and for information about how to apply for permission to reuse the copyright material in this book please see our website at www.wiley.com/wiley-blackwell The right of the author to be identified as the author of this work has been asserted in accordance with the UK Copyright, Designs and Patents Act 1988 All rights reserved No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by the UK Copyright, Designs and Patents Act 1988, without the prior permission of the publisher Designations used by companies to distinguish their products are often claimed as trademarks All brand names and product names used in this book are trade names, service marks, trademarks or registered trademarks of their respective owners The publisher is not associated with any product or vendor mentioned in this book This publication is designed to provide accurate and authoritative information in regard to the subject matter covered It is sold on the understanding that the publisher is not engaged in rendering professional services If professional advice or other expert assistance is required, the services of a competent professional should be sought Library of Congress Cataloging-in-Publication Data Handbook of microalgal culture : applied phycology and biotechnology / edited by Amos Richmond and Qiang Hu – Second edition pages cm Includes bibliographical references and index ISBN 978-0-470-67389-8 (hardback) – ISBN 978-1-118-56716-6 – ISBN 978-1-118-56717-3 (emobi) – ISBN 978-1-118-56718-0 (ePdf) – ISBN 978-1-118-56719-7 (ePub) Algae culture–Handbooks, manuals, etc Microalgae–Biotechnology–Handbooks, manuals, etc Algology–Handbooks, manuals, etc I Richmond, Amos, editor of compilation II Hu, Qiang, 1960- editor of compilation SH389.H37 2013 579.8–dc23 2013006646 A catalogue record for this book is available from the British Library Wiley also publishes its books in a variety of electronic formats Some content that appears in print may not be available in electronic books Cover images: C Amos Richmond and Qiang Hu Cover design by Steve Thompson Set in 9.5/12 pt Times by Aptara R Inc., New Delhi, India 2013 Contents List of Contributors Acknowledgments Introduction vi xiii xiv Part 1: The Microalgal Cell with Reference to Mass Cultures 1 The Microalgal Cell Robert A Andersen Photosynthesis in Microalgae Jiˇr´ı Masoj´ıdek, Giuseppe Torzillo, and Michal Kobl´ızˇek 21 Basic Culturing and Analytical Measurement Techniques Yuan-Kun Lee, Wei Chen, Hui Shen, Danxiang Han, Yantao Li, Howland D T Jones, Jerilyn A Timlin, and Qiang Hu 37 Strategies for Bioprospecting Microalgae for Potential Commercial Applications William Barclay and Kirk Apt 69 Maintenance of Microalgae in Culture Collections Jerry J Brand, Robert A Andersen, and David R Nobles Jr 80 Environmental Stress Physiology with Reference to Mass Cultures Giuseppe Torzillo and Avigad Vonshak 90 Environmental Effects on Cell Composition Qiang Hu 114 Inorganic Algal Nutrition Johan U Grobbelaar 123 Commercial Production of Microalgae via Fermentation William Barclay, Kirk Apt, and X Daniel Dong 134 Molecular Genetic Manipulation of Microalgae: Principles and Applications Roshan Prakash Shrestha, Farzad Haerizadeh, and Mark Hildebrand 146 10 Part 2: Mass Cultivation and Processing of Microalgae 169 11 Biological Principles of Mass Cultivation of Photoautotrophic Microalgae Amos Richmond 171 12 Theoretical Analysis of Culture Growth in Flat-Plate Bioreactors: The Essential Role of Timescales Y Zarmi, G Bel, and C Aflalo 205 iii iv Contents 13 Photobioreactors for Mass Production of Microalgae Graziella C Zittelli, Natascia Biondi, Liliana Rodolfi, and Mario R Tredici 225 14 Downstream Processing of Cell Mass and Products Emilio Molina Grima, Francisco Gabriel Aci´en Fern´andez, and Alfonso Robles Medina 267 15 First Principles of Techno-Economic Analysis of Algal Mass Culture C Meghan Downes and Qiang Hu 310 Part 3: Commercial Species of Industrial Production 327 16 Chlorella: Industrial Production of Cell Mass and Chemicals Jin Liu and Qiang Hu 329 17 Biology and Industrial Production of Arthrospira (Spirulina) Amha Belay 339 18 Dunaliella: Biology, Production, and Markets Michael A Borowitzka 359 19 Biology and Industrial Potential of Botryococcus braunii Makoto M Watanabe and Yuuhiko Tanabe 369 20 Biology and Commercial Aspects of Haematococcus pluvialis Danxiang Han, Yantao Li, and Qiang Hu 388 21 Novel Sulfated Polysaccharides of Red Microalgae: Basics and Applications Shoshana (Malis) Arad and Dorit van Moppes 406 22 Hydrogen Production by Chlamydomonas reinhardtii Giuseppe Torzillo and Michael Seibert 417 23 Biology and Biotechnology of Edible Nostoc Danxiang Han, Zhongyang Deng, Fan Lu, and Zhengyu Hu 433 24 IGV GmbH Experience Report, Industrial Production of Microalgae Under Controlled Conditions: Innovative Prospects O Pulz, J Broneske, and P Waldeck 445 25 Microalgae for Human and Animal Nutrition E Wolfgang Becker 461 26 Bioactive and Novel Chemicals from Microalgae R Cameron Coates, Emily Trentacoste, and William H Gerwick 504 27 High-value Recombinant Protein Production in Microalgae Daniel J Barrera and Stephen P Mayfield 532 28 Molecular and Cellular Mechanisms for Lipid Synthesis and Accumulation in Microalgae: Biotechnological Implications Yantao Li, Danxiang Han, Kangsup Yoon, Shunni Zhu, Milton Sommerfeld, and Qiang Hu 29 Biofuels from Microalgae Maria J Barbosa and Ren´e H Wijffels 545 566 Part 4: Water Pollution and Bioremediation by Microalgae 579 30 581 Eutrophication and Water Poisons Susan Blackburn Contents v 31 Water Purification: Algae in Wastewater Oxidation Ponds Asher Brenner and Aharon Abeliovich 595 32 Absorption and Adsorption of Heavy Metals by Microalgae Drora Kaplan 602 Part 5: Microalgae for Aquaculture 613 33 Microalgae for Aquaculture: The Current Global Situation and Future Trends Arnaud Muller-Feuga 615 34 Microalga for Aquaculture: Practical Implications Oded Zmora, Dan J Grosse, Ning Zou, and Tzachi M Samocha 628 35 Transgenic Marine Microalgae: A Value-Enhanced Fishmeal and Fish Oil Replacement Jonathan Gressel 653 36 Microalgae for Aquaculture: Nutritional Aspects E Wolfgang Becker 671 37 The Enhancement of Marine Productivity for Climate Stabilization and Food Security Ian S.F Jones and Daniel P Harrison 692 Index 705 List of Contributors Aharon Abeliovich† Department of Biotechnology Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel Shoshana (Malis) Arad Professor Department of Biotechnology Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel Email: arad@bgu.ac.il Phone: 972-8-6479069 Fax: 972-8-9479067 † Deceased C Aflalo Jacob Blaustein Institutes for Desert Research Ben-Gurion University of the Negev Midreshet Ben-Gurion 84990, Israel Email: aflaloc@bgu.ac.il Phone: 972-86596817 Fax: 972-86596802 Maria J Barbosa Research Manager Microalgae Food and Biobased Research Wageningen University and Research Center P.O Box 17, 6700 AA Wageningen, The Netherlands Email: maria.barbosa@wur.nl Phone: + 31 (0)317 480079 Fax:+ 31 (0)317 483011 Robert A Andersen Senior Research Scientist Friday Harbor Laboratories University of Washington Friday Harbor, WA 98250 William Barclay retired/former Chief Intellectual Property Officer DSM Nutritional Products – Boulder Research Center 4909 Nautilus Court North, Suite 208, Boulder, CO 80301 Director Emeritus Provasoli-Guillard National Center for Marine Algae and Microbes Bigelow Laboratory for Ocean Sciences P.O Box 380, East Boothbay, ME 04544, USA Email: raa48@uw.edu Phone: 001-906-370-1886 7356 Panorama Drive Boulder, CO 80303, USA Email: bbarclay@icloud.com Phone: 1-303-579-5943 Kirk Apt Chief Research Scientist DSM Nutritional Products 6480 Dobbin Road, Columbia, MD 21045, USA Email: kirk.apt@dsm.com Phone: 1-410-740-0081 Fax: 1-410-740-2985 Daniel J Barrera PhD Student University of California, San Diego 9500 Gilman Drive, La Jolla, CA 92093-0212, USA Email: dbarrera@ucsd.edu Phone: 858-869-3879 vi List of Contributors E Wolfgang Becker Medical Clinic, Department II Immunopathological Laboratory 72076 Tăubingen, Germany Email: ew.becker@gmx.de Phone: 0049-7472-21981 G Bel Department of Solar Energy and Environmental Physics Jacob Blaustein Institutes for Desert Research Ben-Gurion University of the Negev Midreshet Ben-Gurion 84990, Israel Email: bel@bgu.ac.il Phone: 972-86596845 Fax: 972-86596921 Amha Belay Sr Vice President & CTO Earthrise Nutritionals LLC 2151 Michelson Drive, Suite 258 Irvine, CA 92612, USA Email: abelay@earthrise.com Phone: 1-760-427-8462 Natascia Biondi Researcher Dipartimento di Scienze delle Produzioni Agroalimentari e dell’Ambiente Universit`a degli Studi di Firenze Piazzale delle Cascine 24, 50144 Firenze, Italy Email: natascia.biondi@libero.it Phone:+39-055-3288480 Fax: +39-055-3288272 Susan Blackburn Head, Australian National Algae Culture Collection CSIRO Marine and Atmospheric Research G.P.O Box 1538, Hobart, Tasmania 7001, Australia Email: susan.blackburn@csiro.au Phone: +61-3-6232-5307 Fax: +61-3-6232-5000 Michael A Borowitzka Professor Algae R&D Center Murdoch University Murdoch WA 6150, Australia Email: m.borowitzka@murdoch.edu.au Phone: +61-8-9360-2333 vii Jerry J Brand Professor Section of Molecular, Cell & Developmental Biology Email: jbrand@austin.utexas.edu Phone: 512-4711589 Fax: 512-2323402 Director Culture Collection of Algae (UTEX) University of Texas at Austin 205 W 24th Street, Austin, TX 78712, USA Email: jbrand@austin.utexas.edu Phone: (512) 4711589 Fax: (512) 2323402 Asher Brenner Professor Unit of Environmental Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel Email: brenner@bgu.ac.il Phone: 972-8-6479029 J Broneske IGV Biotech Department IGV GmbH Arthur-Scheunert-Allee 40-41, 14558 Nuthetal, Germany Email: j_broneske@igv-gmbh.de Phone: +49-33200-89151 Fax: +49-33200-89158 Wei Chen Associate Research Professor Laboratory for Algae Research and Biotechnology College of Technology and Innovation Arizona State University 7001 E Williams Field Road Mesa, AZ 85212, USA Email: wei.chen@asu.edu Phone: +1-480-727-5663 Fax: +1-480-727-1475 R Cameron Coates Center for Marine Biotechnology and Biomedicine PhD Candidate Scripps Institution of Oceanography University of California, San Diego 9500 Gilman Drive, La Jolla, CA 92093-0212, USA Email: rccoates@ucsd.edu Phone: 01-858-822-4366 Fax: 01-858-534-0576 viii List of Contributors Zhongyang Deng Associate Professor School of Biological Engineering Hubei University of Technology Wuhan, Hubei 430068, China Email: dengculture@yahoo.com Phone: +86-139-0591-4136 X Daniel Dong Sr Research Engineer IV DSM Nutritional Products 6480 Dobbin Rd, Columbia, MD 21045 Email: Daniel.dong@dsm.com Phone: 1-240-512-2643 Fax: 1-410-740-2985 C Meghan Downes Associate Professor Economics, International Business, and Applied Statistics Department New Mexico State University Las Cruces, NM 88003-8001, USA Email: cdownes@nmsu.edu Phone: 575-202-5181 Francisco Gabriel Aci´en Fern´andez Associate Professor of Chemical Engineering Chemical Engineering Department University of Almer´ıa 04120 Almeria, Spain Email: facien@ual.es Phone: +34-950015443 Fax: +34-950015484 William H Gerwick Distinguished Professor Center for Marine Biotechnology and Biomedicine Scripps Institution of Oceanography Skaggs School of Pharmacy and Pharmaceutical Sciences University of California, San Diego 9500 Gilman Drive, La Jolla, CA 92093-0212, USA Email: wgerwick@ucsd.edu Phone: 01-858-534-0578 Fax: 01-858-534-0576 Jonathan Gressel Professor Emeritus Department of Plant Sciences Weizmann Institute of Science Rehovot 76100, Israel Email: jonathan.gressel@weizmann.ac.il Phone: +972-8-9343481 Fax: +972-8-9344181 Daniel J Grosse President TerrAqua Environmental Science and Policy, LLC 3754 Jenifer Street, NW Washington, DC 20015 Email: dgrosse@terraqua.org Phone: 202-258-9700 Fax: 202-244-4667 and Adjunct Associate Professor University of Maryland University College Graduate Program in Environmental Management Adelphi, MD 20873, USA Email: Dan.Grosse@faculty.umuc.edu Phone: 202-258-9700 Fax:202-244-4667 Emilio Molina Grima Professor of Chemical Engineering Chemical Engineering Department University of Almer´ıa 04120 Almeria, Spain Email: emolina@ual.es Phone: +34-950015032 Fax: +34-950015484 Johan U Grobbelaar Professor Emeritus Department of Plant Sciences University of the Free State Bloemfontein 9300, South Africa Email: grobbeju@ufs.ac.za Phone: +27-51-4012263 Farzad Haerizadeh Metabolic Systems Lead Synthetic Biology R&D Life Technologies Corporation 5791 Van Allen Way Carlsbad, CA 92008, USA Email: farzad.haerizadeh@lifetech.com Phone: 760-476-6156 Danxiang Han Assistant Research Professor Laboratory for Algae Research and Biotechnology College of Technology and Innovation Arizona State University 7001 E Williams Field Road Mesa, AZ 85212, USA Email: Danxiang.han@asu.edu Phone: +1-480-727-5661 Fax: +1-480-727-1475 List of Contributors Daniel P Harrison Research Engineer Ocean Nourishment Foundation P.O Box 363, Glebe 2037 NSW, Australia Email: daniel.harrison@onf-ocean.org Phone: +61-409-398-901 Mark Hildebrand Research Scientist Scripps Institution of Oceanography University of California San Diego, CA 92037, USA Email: mhildebrand@ucsd.edu Phone: 858-822-0167 Fax: 858-534-7313 Qiang Hu Professor and Co-Director Laboratory for Algae Research and Biotechnology College of Technology and Innovation Arizona State University 7001 E Williams Field Road Mesa, AZ 85212, USA Email: huqiang@asu.edu Phone: +1-480-727-1784 Fax: +1-480-727-1475 Zhengyu Hu Professor Institute of Hydrobiology Chinese Academy of Sciences South Donghu Road, Wuhan, Hubei 430072, China Email: huzy@ihb.ac.cn Phone: +86-138-0864-8218 Fax: +86-27-6878-0016 Howland D.T Jones Senior Scientist Biosciences Sandia National Laboratories P.O Box 5800 Albuquerque, NM 87185, USA Email: hdjones@sandia.gov Phone: +1-505-284-1842 Fax: +1-505-284-3775 Ian S.F Jones Professor Ocean Technology Group School of Geosciences University of Sydney Madsen Building F09, 2006 NSW, Australia Email: ian.jones@sydney.edu.au Phone: +61-2-9351-4585 ix Drora Kaplan Professor Emeritus, Department of Environmental Hydrology and Microbiology Zuckerberg Institute for Water Research Jacob Blaustein Institutes for Desert Research Ben-Gurion University of the Negev Sede Boqer Campus, Midreshet Ben-Gurion 84990, Israel Email: droraka@bgu.ac.il Phone: 972-8-6596835 Fax: 972-8-6596909 Michal Kobl´ızˇ ek Senior Scientist Department of Phototrophic Microorganisms Institute of Microbiology Academy of Sciences Opatovick´y ml´yn, CZ-37981 Tˇreboˇn, Czech Republic Email: koblizek@alga.cz Phone: +420-384-340432; Fax: +420-384-340415 Yuan-Kun Lee Associate Professor Department of Microbiology Yong Loo Lin School of Medicine National University of Singapore Block MD4, Science Drive 2, Singapore 117597 Email: micleeyk@nus.edu.sg Phone: +65-65163284 Fax: +65-67766872 Yantao Li Assistant Research Professor Laboratory for Algae Research and Biotechnology College of Technology and Innovation Arizona State University 7001 E Williams Field Road Mesa, AZ 85212, USA Email: yantao.li@asu.edu Phone: +1-480-727-5662 Fax: +1-480-727-1475 Jin Liu Faculty Research Associate Laboratory for Algae Research and Biotechnology College of Technology and Innovation Arizona State University 7001 E Williams Field Road Mesa, AZ 85212, USA Email: gjinliu@gmail.com Phone: +1-480-727-1410 Fax: +1-480-727-1475 Index Note: Page numbers in italics refer to Figures; those in bold to Tables abalone aquaculture 641–42, 688–9 absorbance colorimetry for total carbohydrates 49–50 measurement for chlorophyll estimation 41–2 see also optical density absorption coefficient (radiation intensity in reactors) 189, 209 of heavy metals, tolerance mechanisms 604, 604, 605 optical cross section calculation 92, 94 of photon energy, mathematical models 209–12 accelerated solvent extraction (ASE), lipids 55, 57 accessory pigments, function 25 acclimation see light acclimation Accordion unit PBR design 237 acetyl-CoA carboxylase (ACCase) 524, 545–6, 558 acidification, ocean 694, 700 acidophilic microaglae, collection strategies 71–2 activated sludge process 597 acyl transferase enzymes (lipid biosynthesis) 547–9, 549–50, 550–1 adsorption of cationic polymers, in flocculation 269 expanded bed, extraction method 292 of heavy metals by algal exopolymers 604, 604–5 aflatoxin-B1 505, 506 agar media 83 aggregation ability, in ‘sticky’ strains 73 for algal harvesting 268–71 caused by contaminants 635 Agrobacterium tumefaciens, as transformation agent 150–1, 536 algae bioprospecting for commercial potential 70 defining features diversity in culture collections 81 education and training resources 89 range of nutritional types 123–4, 124 Algae Growth System (AGS) flexible film PBRs 239–41, 240 Algae Tunnel hybrid PBR system 243, 244 AlgaePARC (Wageningen) pilot facility 571, 572 algal blooms frequency and dangers 482, 582, 582–3 in green water polyculture 617–18 from incubation of samples 38 management 674 natural, harvesting exploitation 227 vertical movement in water column 587 see also harmful algal blooms alginates, for immobilized culture systems 48–9 alkaline cell lysis 282 Allen’s culture medium, composition 126, 128, 131 allergic reactions, inhibition by Spirulina 492 amino acids 468–71, 470, 508, 662–3, 676 ammonia, in wastewater 598 amnesic shellfish poisoning (ASP) 586 amoeboid algae 5–6, 6, 16 anaerobic ponds (waste treatment) 597 animal feed algae and conventional feeds compared 486–8 algal concentrates for aquaculture 634-5, 641, 685 algal diets for toxicity testing 484–5 colourant additives 480, 481, 684 enzyme and mineral additives 664–5 fishmeal benefits and demand 653–4, 654, 662 uses of Chlorella 334, 632–3 annual cycles see seasonal cycles annular column photobioreactors 229–30, 230 antennae, light-harvesting in cyanobacteria, response to light levels 96 energy transfer 26, 28 pigments 25, 26 small size and light acclimation 104, 567 truncation in C reinhardtii mutants 421 anti-inflammatory activity 438, 484, 510–11 antibiotics for culture purification 38 effects on Porphyridium growth 409, 410 resistance, selection markers 149, 657 transgenic antibacterial algae used as alternative 159, 665 anticancer activity antitumour products 39, 437, 438, 484, 489 carcinogen sequestration, chlorophyllin 505 β-carotene, range of action 506 functional genomic investigation 515–16 microalgal cytotoxins 511, 511–12 antimicrobial peptides 659, 665 antisense RNA gene knockdown 154 antiviral agents 412, 437–8, 491–2, 508, 508 aquaculture algal feed compared with fishmeal 54–5, 662, 666 clear water, microalgal requirements 618–22, 620 costs of algal mass cultivation systems 622, 623, 631 global production statistics 616, 616–17, 617 Handbook of Microalgal Culture: Applied Phycology and Biotechnology, Second Edition Edited by Amos Richmond and Qiang Hu C 2013 John Wiley & Sons, Ltd Published 2013 by Blackwell Publishing Ltd 705 706 aquaculture (Continued ) green water culture techniques 616, 617–18, 635–6 integrated (sustainable) schemes 645 market value enhancement of produce 622–3 risk of algal bloom stimulation 589, 672 shellfish, toxin levels 588–9 species used in feed 628–9, 629, 673, 674 use of dry formulated feeds 621, 622, 623 use of microalgal biofuel residues 226, 257 Aquatic Species Program (US) 74 aqueous extraction processing (AEP) 290–1 aqueous two-phase systems (ATPS) 290 areal productivity and cell density 174 compared with alternative metrics 255, 313 and mixing 178 and optical path length 181, 181, 181–2 in reactor efficiency assessment 195–6, 196 Artemia (brine shrimp) as by-product of Dunaliella culture 365 growing methods for aquaculture feed 644, 674 specific aquaculture uses 672 Arthrospira (Spirulina) animal feed uses 486, 487–8 aquaculture uses 687–9 carotenoid spectrum 478, 479 ecology in African soda lakes 342–3 economic analysis of raceway pond production 321–4, 323–4 fatty acid composition 473, 474, 475 industrial mass culture production 346–54, 348, 353–4 morphology and ultrastructure 340–2, 341 physiology 343–6 product composition and quality 352, 352–3, 353 taxonomic status and research 339–40, 463 toxicology studies 484, 486 traditional food uses 461–2 vitamin B12 bioavailability 475–6 Zarrouck’s culture medium 126, 128, 345, 349 ascorbic acid (vitamin C) early commercial fermentation production 137 enrichment in rotifers from algal feed 674 levels in algal species 680 asexual reproduction autospore production cycle (Chlorella) 330 Index implications for biotechnology 8, 159, 655 vegetative growth cycle (Haematococcus pluvialis) 389–90 ash content 41, 313 assessment economic factors 197, 297, 310–11 life cycle (LCA), for production impacts 382, 427, 573–4 reactor productivity and efficiency 195–7, 197, 256–7 see also techno-economic analysis astaxanthin biosynthesis effect of stress factors 118 enzymes and pathways 391–3, 392 physiological role 395, 395 nutritional benefits for fish and shrimp 684 structure and uses 388–9, 480, 507, 683–4 asthma, effects of β-carotene 491 ATP synthesis, enzyme complex 26, 28–9 attached microalgae isolation of strains from rocks 71 photobioreactor designs 246–7, 254, 257 attenuation coefficient 189, 209 auto-flocculation 271, 272 autoinhibitors 124, 183–5, 184 automated cell counting see flow cytometry autotrophy anoxygenic 21 categories, in algae 123 nutrient requirements 125 auxotrophy 124, 477 average cell travel time 187–8 average radiation intensity 189–90, 192, 192, 221 avermectin insecticides 659 axenic cultures collection of suitable strains 71–2 design of PBRs 247–8 maintenance in culture collections 83, 86, 88 purification techniques 38 azaspiracid (AZA) 587 backup cultures (in collections) 86 bacteria as bioflocculation agents 270 contamination of algal products 496, 635 fecal coliforms 598 photosynthetic 21 separation of microalgae from 38, 85–6 barcoding 88 batch culture growth rate phases 43–5, 44 productivity, compared with continuous flow 47, 316, 641 for rotifer production 643–4 BCA (bicinchoninic acid) protein assay 52 bead mills, for cell disruption 283 benthic microalgae for abalone aquaculture 642 culture media 83 photobioreactor designs 246–7 BG-11 culture medium, composition 126, 128, 131, 375 Bigogno analysis method (lipids) 54 bio-rational collection–screening process 75–7, 76, 139 bioactivity compounds from Nostoc species 437–8 lipid fatty acids 473, 475 research on novel red algal products 411–12 screening by bioassays 39, 513–14, 514 studies on animals and humans 484–6 biochemical composition see chemical composition analysis biochemical oxygen demand (BOD) 597, 599, 645 biochemical products diversity, from algal sources 11–12, 507–8, 512 extraction and purification methods 284–6, 288–93, 303 proteins and metabolic engineering 156–9, 157–8 screening of microalgae for 39, 72, 513–14, 514 shelf life 351 biocoils (helical photobioreactors) 234, 632 biodegradable plastics 524, 524–5 biodiesel biorefinery plant, concept design 571–3, 573 collection of potential algal strains from seawater 71 compared with petroleum product 369, 599 expense of waste treatment 600 heterotrophic microalgal production 139–41, 142 market value and demand 566, 599–600 production processes 293–7, 294 biodiversity extent and scope in algae 12 in microalgal culture collections 81 bioflocculants 270, 271 biofuels advantages of using microalgae 418, 428, 462, 565–6 biosynthetic manipulation of lipid metabolism 523–4 carbon feedstocks 140, 141 coproducts from production process 226, 504, 512 Index microalgal, economic viability 226, 298, 303, 369, 566 octane/cetane rating 522–3, 523 research and development needed 566–74 research on potential of Chlorella 334–5 see also biodiesel biogenic toxins 482–3 biolistic (particle bombardment) transformation 149–50, 151, 394, 536 Biological Algae Growth System (BAGS) hybrid PBR 242 biological value (BV), proteins 467–8, 469 biomass for biosorption of heavy metals 606–7 energy content 33 fractionation, in biorefinery 571–3 loss at night 198–9 measurement of production 31, 39–43, 45–7, 313–16 nonliving algae as products 606, 635, 685 produced from fermentation 135, 136 production costs, biodiesel case study 297, 297–8, 298–9 toxicological testing in animals 484–5 see also productivity bioprospecting multivariate approaches 73–7, 76 univariate strategies 70–3 biorefineries 571–3, 573 bioremediation algal proteins, scope for metabolic engineering 156, 157 applications for Chlorella cultures 335 capacity of microalgae for 131, 605–7 plant-based, algal, and mechanical systems 595–6 see also wastewater treatment biosensors, microalgal 603 biotechnology see genetic engineering biphasic culture media 83 bivalves see molluscs Blackman model, light response curve 92, 93, 98, 98 Bligh–Dyer analysis method (lipids) 54, 288, 291 blue-green algae see cyanobacteria BODIPY (neutral lipids dye) 63 Bold’s Basal culture medium composition 126, 128, 131 use and modification for Haematococcus culture 396, 397 Botryococcus braunii culture media 374–5, 375 effects of culture conditions 375–8, 377 hydrocarbon biosynthetic pathways 521 race A 372, 373 race B 372–4, 374 mass culture approaches 378–80, 380 natural occurrence and hydrocarbon production 369–70, 370 oil production, business evaluation 380–4, 381, 382, 383 phylogeny and taxonomy 370–2, 371 Bradford assay methods (protein) 52–3 breeding, challenges in microalgae 159–60, 655 brevetoxins 507, 509, 509–10, 586 broth media 82–3 Brownian motion 207 buffer systems 125–6, 345 business analysis see techno-economic analysis calcium alginate beads 48–9 Calvin–Benson cycle 29, 29–30, 378 canthaxanthin 684 capital costs (CAPEX) biodiesel case study 298, 299, 301 in techno-economic analysis 317, 318, 319–20, 321 carbohydrates accumulation under nitrogen depletion 116 algal storage product types 10, 11–12, 551–2 amount related to cell density 177 digestibility 471, 487, 662 production in high light conditions 115 quantitative measurement 49–51 see also sulfated polysaccharides carbon dioxide see CO2 carbon fixation Chlorella as model organism for study 462 collection of suitable strains 73–4 measurement techniques 31 and pH rise 125–6 reaction pathways 29, 29–30, 522, 522 removal of ability (transgenic mitigation) 661 sequestration by marine algae 693, 698 Carbon Recycling Facility (CRF) film membrane PBR 247 CARET (β-carotene and retinol efficacy trial), lung cancer 491 β-carotene as astaxanthin precursor 391 commercial producers and market value 365 Dunaliella as commercial source 359, 364, 478 health benefit assessment 364–5, 479, 491, 506 isomers in natural and synthetic products 361, 364, 480 toxicity testing 485 707 carotenogenesis biosynthetic pathways 391–3, 392 chemical changes under stress 391, 478 effects of environmental conditions 118, 361, 361, 363 genetic regulation 393–4 ultrastructure changes in H pluvialis 390, 390–1 carotenoids accumulation and storage in Dunaliella 360, 361, 478 algal components and products 478–81, 479, 506 chemical structure and roles 24, 25, 478, 506 composition analysis, aquaculture algae 681, 681–85, 682 composition used for Nostoc classification 436 diversity in heterokont clades 15 high-production strains, collection and screening 71, 481 market value 507, 683 products from Chlorella spp 334 carp, freshwater aquaculture 688 cascade culture systems 228, 228–9 cell concentration (density) cell number counting methods 39–40 control in aquaculture hatcheries 638, 643 and control of biochemical composition 118–19 effect on light penetration depth 176, 177 effects on cell ultrastructure and composition 177 mutual shading and light–dark cycles 173, 207 optimum (OCD), for maximal yield 173–6, 175, 199, 208–9 quorum sensing (sudden culture decline) 658 related to average radiation intensity 189–90 cell recycled culture system 48 cell walls in Arthrospira (Spirulina) 341 of diatoms, fine structure 7–8, disruption methods, for chemical extraction 50, 58, 466 diversity in algae 11 heavy metal exclusion mechanisms 604, 604 polysaccharides digestibility 662, 664 in Nostoc 437–8 in red microalgae 407–8, 408–9 secondary development in for H pluvialis 390, 390–1, 394 thickness and ease of extraction 571 708 cells cycles in favourable and stress conditions 389–90 disruption methods 281–4, 282, 303, 573 division 11 mammalian, serum-free culture 492, 532 motion, in reactor turbulent flow 187–8 random motion 207–8, 209, 220 structure in microalgae 8–11, washing technique 38 centrifugal recovery from culture media 274–6, 277–8, 287 chelating agents in algal dells, for heavy metals 604 in animal feed additives 664 in culture media 127 chemical cell disruption methods 282, 466 chemical composition analysis algal species used in aquaculture 674, 675 amino acids 468–71, 470, 676 ash and moisture content 41, 463 carbohydrates 49–51, 471 chlorophyll estimation 41–2 comparison of algal taxonomic groups 677, 679 effect of environmental conditions 114–16, 331 fatty acid methyl esters (FAME) 61–2 lipids 53–61, 62–4, 471–5, 676–80 pigments 477–82 proteins 51–3, 463–8, 674–6 range, algae compared with other foods 463, 464–5 total organic carbon (TOC) 42, 114 toxins 482–3 vitamins 475–7, 476, 680–81 chemical score (CS), proteins 469 chemostat culture systems 47, 399 chitosan (flocculation agent) 270, 270, 271 Chlamydomonas reinhardtii chloroplasts, recombinant protein production 533 cultivation in photobioreactors 422–6, 425 culture contamination risks 428 effects of sulfur deprivation 422, 423 hydrogen photoproduction 417–18, 419 mutant strains 419, 421 nuclear transformation stability 657 chlorarachniophytes 14, 16 Chlorella animal/human testing for bioactivity 484, 485, 490 autotrophic mass cultivation 331, 333 carotenoid spectrum 478, 479 composition and physiology 330–1, 332 contamination of Spirulina cultures 200 fermentation production systems 137, 139–41, 331, 333 Index harvesting and drying 333–4 history of commercial cultivation 136, 331, 454 nutritional quality for rotifer feeding 632–3, 677 potential products and applications 329, 334–6 Sorokin–Krauss culture medium 126, 128 species characteristics 330 Chlorella growth factor (CGF) 334 chlorellin, antibiotic activity 183, 489 Chlorophyceae, taxonomy 370–1 chlorophyll fluorescence technique parameters calculated from measurements 33, 93–4 photochemical principles 31–2, 32 for in situ culture performance monitoring 198 used to monitor photoinhibition 99–100, 102 chlorophyllin 505–6, 506 chlorophylls amount, related to cell density 177 and astaxanthin synthesis 391 chemical structure and roles 23–5, 24, 505, 506 health effects of consumption 477-8, 505–6 interference in protein analysis 52, 53 quantity determination 41–2 chloroplasts envelope, formation of lipid bodies 554, 554–5, 555 fatty acid elongation in 372, 546 for recombinant protein production 533 ultrastructure 8–10, see also thylakoids cholesterol reduction, bioactive algal products 437, 438, 473, 490, 493 chromatography elution methods for product extraction 292, 292–3 lipids analysis methods 55–8, 57, 60–2 as step in new product discovery 513, 514 chytrids (fungal contaminants) 201, 399–400 ciguatera fish poisoning (CFP) 587 circulation pumps 448 clams, aquaculture 640–41, 686 climate stabilization 694 CO2 (carbon dioxide) atmospheric increase, anthropogenic 692 effect on cell wall growth, red algae 408 emissions EU greenhouse gas targets 565 industrial scrubbing systems 70–1, 73–4, 335, 694 life cycle assessment, production plants 382, 384 flow feed equipment 448 solubility limitation in high-salinity brines 360–1 supercritical, in extraction of products 291–2, 293 supply control in closed PBR design 457–9 supply for aquatic algal production 125–6, 439, 568 coagulation 268–9, 276–7 see also flocculation coccoid algae 5–6, Codex Alimentarius Commission (CAC) 353, 495 codon usage, transgenic algae 154–5, 536–7, 537 collection techniques broad-based strategies 70–1 habitat-specific 71–2, 74 sources of microalgae 37, 72–3 strategies for strains with multiple traits 73–7 collections (microalgal species/strains) 80–1, 88–9, 125, 462, 584 colonial algae cell counting 39 growth forms 4, 5–6, 6–7 Nostoc macrocolonies 434, 435, 436 column chromatography, lipids analysis/purification 56–8, 57, 293 commercial applications goods and services from culture collections 80–1, 584 history of fermentation technologies 136–7 range of successful microalga technologies 69, 225–6 producers for aquaculture feed supply 633-4, 633, 634 using fermentation 137–9, 138 world Spirulina production 346–7 trends in technological development 456 contamination avoidance in culture collections 86 control in PBR hydrogen production 428 of environment by transgenics 496 fungal parasites 399–400, 659 invading species, competition for limiting nutrients 130 monitoring and control in outdoor culture 200–1, 350, 635 resistance by transgene engineering 658–60 continuous flow culture systems methods 45, 45–8, 293–6 for rotifer production 644 convective hot air drying 280 conversion factors, units 313, 314–15, 316 copper toxicity, alleviation in fish 688 Index cost-effectiveness see economic considerations crabs, aquaculture 637 crop protection see contamination crustaceans see crabs, aquaculture; shrimps cryopreservation 86–7 Crypthecodinium fermentation, for DHA 137–8, 138, 141–2 cryptophytes 15–16 cultures, microalgal conditions and containers for culture collections 83–4, 85 cultivation systems 43–9, 252–4 isolation and purification 37–8 media types and components 82–3, 124–5, 126, 128 see also maintenance of cultures; mass culture cyanobacteria in culture collections 81 culture purification with antibiotics 38 effects on ancient atmosphere 12–13, 521 genetic engineering suitability 658 iron-stress-induced proteins (isiA) 117 lipid metabolism 471, 518, 521 photosynthetic pigments 26, 96 physiology 13, 344, 345, 346 secondary metabolite therapeutic products 508, 510–11, 512 species-diagnostic molecular markers 434–6 ultrastructure 8, 9, 21, 341–2 cyanophycin granules 341–2 cyanotoxins 586–7 cyclostat culture systems 47, 48 cytochrome b6 /f complex 27, 27–8 dairy wastewater 599 dark reactions (of photosynthesis) 22, 22, 29, 29–30 dark volume residence time 188–9, 190, 207, 215 data sources, for economic analysis 318–19, 319 dehydration effects on protein quality 467, 468 methods, algal biomass 278–81, 301, 303, 467, 467 methods for Arthrospira (Spirulina) 351 tolerance of dessication, Nostoc spp 438, 439 used in Chlorella mass cultivation 334 density gradient centrifugation 38 desert areas environmental conditions and photoinhibition 102–3 suitability for Arthrospira (Spirulina) culture 347 water resources assessment 574 detoxification (heavy metals) applied remediation methods 605 biological mechanisms 604, 604–5 DGAT (diacylglycerol acyltransferase) 549–50 DHA (docosahexaenoic acid) fermentation production with microalgae 137–9, 138, 140 food and animal feed markets 139, 318, 473 polyketide synthase (PKS) synthetic pathway 136, 518 diarrhetic shellfish poisoning (DSP) 586 diatoms as aquaculture feed 638, 640, 642 biofilm polycultures 247 early mass cultivation attempts 462 morphological diversity 7, 7–8, silicon requirement 128 use of gene expression control elements 147 dietary supplements see nutritional supplements digestibility, whole algae 642, 654, 662 digestibility coefficient (DC), proteins 468, 469 ‘digestion’ period, photosynthetic cycle 220, 222 dih´e (traditional Spirulina food) 346, 461–2 dilution (light), reactor design for by turbulent mixing 180–1 by vertical/inclined arrangement 195, 426, 567, 567 dilution rate (D) 46, 46 dinoflagellates characteristics 14, 15 ion channel modulating products 509, 509–10 mass cultivation challenges 248 toxic blooms 583 discounted cash flow (DCF) analysis 383, 383 dispersed air flotation 273, 276, 276, 333–4 disposable panel culture chambers 237 dissolved organic matter (DOM) 350, 351 dissolved oxygen (DO) diurnal cycle 191, 622 measurement and monitoring 198 during sulfur starvation 422, 423 diurnal cycles dinoflagellate vertical migration 587, 588 photon flux density 190 photosynthetic activity 99–100, 100 related to cell density and photoinhibition 190–3, 191 related to hydrogen production 424, 425 DNA barcoding 88 docosahexaenoic acid see DHA 709 doubling time (td ) 42, 47, 396, 441 downstream processing biomass yield losses 313 cell disruption 281–4 dehydration of biomass 278–81 effects on nutritional quality 465–7 multiple methods and pretreatment 400 product extraction 284–6, 288–92 product purification 292–3 DOXP pathway 391, 392 draft oven drying 280 drinking water quality, algal toxins 589 dry weight measurement 40, 40–1, 313 drying see dehydration Dunaliella animal feeding studies, for toxicity tests 478, 485 Ben-Amotz & Avron’s culture medium 126, 128 bioactivity of alga and products 364–5 commercial producers and by-products 365 D salina production technology 361–4, 362 distribution and characteristics 359–60, 360 salt tolerance and physiology 360–1, 361 ecology in African soda lakes 342–3 algae/bacteria in waste stabilization ponds 600 biogeochemical cycles, oceans 693–4, 698 coastal pools with variable salinity 362 ecosystem effects of harmful algae 582, 585–6, 587 marine productivity and food webs 694–5, 701 metals, aquatic ecosystem interactions 602–3, 603 nonsterile outdoor reactors, as ecological niche 200–1 occurrence and productivity of algae 16–17 risks of transgenic spills 661–62 roles of secondary metabolites 507 economic considerations carbon trading financial resources 694, 695, 699 causes of high production costs, Nostoc 441 costs of ocean nitrogen fertilization 699, 701 energy consumption 381, 382, 567–8 equipment costs 70, 317, 318 funding for public service collections 88 importance in assessing viability 310–11, 324–5 710 economic considerations (Continued ) in reactor efficiency assessment 197, 256–7, 459 and trophic independence, in aquaculture 623 see also techno-economic analysis effluent, industrial, as growth medium 379 Elbingerode (Germany) industrial facility 455, 456 electrocoagulation 276, 276–7 electromagnetic radiation, spectrum 22–3, 23 electron transport rate (ETR), calculation 94 electroporation, for genetic transformation 150, 150, 151, 536 endoplasmic reticulum 10, 390, 546–7, 553–4 endosymbiosis, events in algal evolution 12, 13, 14, 16 energy requirements European strategic plans 565 input/output balance in biofuels operations 567–8, 574 for metabolic maintenance 43, 46, 46 world demand and resources 417, 462–3, 504 enrichment methods and media 37–8, 72 sequential/simultaneous, for multiple characteristics 73–5 ensemble-averaged kinetics photosynthetic models 210–12 entrapment, for immobilized cultures 48–9 environmental conditions changes and stress responses 90, 107 cycles, for outdoor algal cultures 91 effects on growth of Haematococcus pluvialis 396 effects on microalgal chemical composition 114–16, 361 required for Arthrospira (Spirulina) culture 347 synergistic effects of multiple factors 97–106, 118, 131 enzyme supplements, animal feed 664–5 EPA (eicosapentaenoic acid) 473 equipment costs dependence on scale of activity 453, 453 as element of CAPEX in analysis 317, 318 microalgal biodiesel production 298, 300, 300 ESI (electrospray ionization)–MS 58, 59, 60 essential amino acid index (EAA) 469, 471 ethanol, microalgal production systems 242–4, 243 euglenoids 16 Index eukaryotic algae evolutionary origins 13 genetic diversity in collections 81 lipid bodies 553–6, 554 phylogenetic super groups 13–14 eutrophication anthropogenic causes 582 control by integrated aquaculture systems 645 definition 581 ecosystem impacts 584, 589–90 risk minimization, ocean nourishment 700 evolution development of major algal groups 13–16 earliest origins of algae 12–13 of physiological processes 21 speed, in microorganisms 81 expanded bed adsorption 292 exponential growth phase 43–4, 44, 631 extraction see isolation extremophilic microalgae collection 71–2 tolerance of hypersaline habitats 359, 360–1 exudates (excreted organic compounds) 124, 183–4, 604 eyespots (stigmata) 11 f/2 medium, aquaculture hatchery algae 630-31 factor cancel method (FCM) 313, 316 facultative ponds (waste treatment) 597 FAME (fatty acid methyl esters) analysis 61–2 transesterification and recovery for biodiesel 296, 296–7, 522 fatty acid synthase (FAS) 546 fatty acids antibacterial activity 674 bioconversion to hydrocarbons 518–21, 520 biosynthetic pathways 545–7, 548 chain length and saturation 546–7 composition (profile) analysis 61–2, 436, 472, 473, 474 content, effect of salinity 376 crosstalk with astaxanthin synthesis 394 oxygen needs in biosynthesis 135–6 in degradation 599 profile in aquaculture feed species 677, 678 very long-chain (VLCFAs), from Botryococcus 372, 373 fed-batch culture system 48, 49 fence-like (‘biofence’) PBR design 233–4, 234 Fenton reaction 117, 396 fermentation available technological capacity 134–5 early commercial production of microalgae 136–7 media and microalgal metabolic pathways 135–6 successful microalgal products 137–9, 138, 333 fermentors bubble-column, for microalgal oil production 137, 138 compared with outdoor cultivation systems 135, 445 corrosion risk for marine microalgae 139 for PUFA-enriched mariculture algae 634 ferredoxin (Fd) 418, 419 fertilization, oceans 694, 695–9, 697 fibromyalgia syndrome 489–90 filamentous algae morphology 5–6, 6–7, 340–1 use of trichomes in mass cultivation 351, 440 filter feeding, aquatic animals 628, 645, 686–7 filtration for culture purification 38 harvest efficiency and culture damage 351 methods in cell harvesting 273–4, 276, 287 financial aspects of production see economic considerations fish algal colouring additives 334, 389, 684 carotenoid metabolism 682 dangers to, from harmful algal blooms 582, 586 drug and growth regulator delivery 665 farming and Arthrospira (Spirulina) feed 687–9 oils, omega-3 fatty acid content 517, 518, 654, 676 production estimates, with ocean fertilization 695–6 trophic level 621, 623 fisheries hatchery production, marine finfish 622, 636–7 use of microalgae for rotifer feed 620, 621–22 world production trends 616, 616, 618 fishmeal: production, demand, and economic value 653–4, 654 flagellates astaxanthin synthesis in motile stages 399 flagella, ultrastructure and motion 10–11 loss of flagella, for transgenic mitigation 661–62 morphological diversity 4, flashing light regimes 213, 213–15, 216, 222 Index flat panel airlift (FPA) PBR design 235, 236 flat plate bioreactors compared with tubular design 294, 333 geometry and design 206, 207, 234–6, 235 mesh ultra-thin layer (MUTL) design 456–7, 457, 458, 459 optical path length 181–2, 207, 208, 410 performance assessment 294, 295 thin, compared with pond/raceway bioreactors 205–7 tilt angle 194, 194 flexible film panel photobioreactors 236–41, 238, 240, 253, 457 floating photobioreactor systems 244–6, 245, 254, 624–5, 625 floating pond PBRs 246 flocculation auto-flocculation, pH dependence 271, 272 definition and theoretical principles 268–9 energy advantage for algal harvesting 570–1 methods and uses in Chlorella mass culture 333 using bioflocculants 270 using metal salts and polyelectrolytes 269–70, 270, 271 flotation, for algal separation 271–3 flow cytometry (FCM) 39–40, 74, 160, 540 fluorescence of chlorophyll, for monitoring photosynthesis 31–2, 32, 33 labelling, for selection/diagnosis markers 160, 482, 507, 540 spectroscopy/microscopy for lipids detection 62–4, 63 fluorescent lamps, for culture collections 83–4 folate, high-production strains, isolation from oligotrophic pools 71 Folch analysis method (lipids) 53–4, 58 food approval of new products for consumption 463 colouring additives 334, 479, 683 ingredients from microalgae heterotrophic production 139, 141, 142 potential for Spirulina 354, 462 traditional uses of algae 346, 433–4, 437, 461–2 food security as driver of new agrochemicals market 513 forecasts of food scarcity 462, 624, 693, 694 supplements to combat nutritional deficiencies 354 four-steps absorption photosynthetic model 211, 211–12, 216–17 freeze-drying (lyophilization) 280–1, 467, 634, 686 freshwater microalgae diversity of types in culture collections 81 extensive pond aquaculture 645 in saline/alkaline lakes (Africa) 342–3 strain isolation from oligotrophic ponds 71 fungi, parasitic contaminants 201, 399–400, 659 gas chromatography (GC) analysis 61–2 gas vesicles (microalgal cells) 342, 587 gene silencing 154 genes eukaryote, structure and expression 146–7, 147 mRNA control elements in microalgae 147–9, 148 targets for metabolic engineering 156 upregulation in stress response 393–4 genetic engineering breeding and selection as alternative 159–60 enhancing heterotrophic capabilities 142 enhancing value/quality as feed meal 662–5 extremophilic algae as gene sources 72, 658 gene stacking 659, 660–61 gene targeting and recombination 151–4, 153 manipulation of algal metabolism 155–9, 157–8, 394, 569–70 potential benefits 354, 655 prevention of GMO environmental release 247–8, 661–62 problems in algal transgenics 154–5, 656–7 red microalgae 409–10, 412 for resistance to contamination 658–60 risks of use, and regulations 496, 699-700 transformation in chloroplasts 155 transformation methods 149–51, 150, 533, 536 genomes cryptic endosymbiosis 12, 14 functional investigation of bioactivity 515–16 sequencing for identification 87–8, 160, 515 for new biosynthetic product potential 156, 514–15, 570 transcription and translation 146–7, 147 GGGT (galactolipid:galactolipid galactosyltransferase) 550 GLA (γ -linolenic acid) 473, 475, 488 711 glass beads transformation method 150, 151, 533, 536 glaucophyte algae 13, 14 glycerolipids in lipid body membranes 552–3 metabolic pathways in Botryococcus braunii 372, 373 quantitative analysis using MS 58–61, 59, 62 see also triacylglycerols GMOs (genetically modified organisms) see genetic engineering Golgi bodies 10, 408–9 good manufacturing practices (GMPs) 352–3, 495 GPAT (glycerol-3-phosphate acyltransferase) 547–9 GRAS (Generally Recognised As Safe) status 159, 353, 495, 532 gravitational separation methods 271–3 gray water see wastewater treatment grazers (algal culture contaminants) 201, 635 green algae 13, 14 green photic volume 176 Green Wall Panel (GWP) reactors 237–8, 238 green water aquaculture benefits 635–6, 674 in fish rearing 622, 645 inorganic substitutes for algae 636 principles and world distribution 616, 617–18 shrimp hatcheries 620 greenhouse gases 694, 701 Grower HarvesterTM PBR design 247 growth hormone, transgenic 665 growth parameters biomass determination 40–2 cell count methods 39–40 in continuous flow cultures 45–7, 46 effect of cell concentration 174 importance of monitoring 198 rate and yield measurements 42–3 growth phases, in batch culture 43–5, 44 growth rates, influencing factors accumulation of inhibitory substances 183–5 irradiance 91, 97, 171–3 nutrient concentrations 128–31, 129 oxygen concentration 102–3 temperature 100–1, 116 turbulence 125, 180 Haematococcus pluvialis biosynthesis of astaxanthin, pathways 391–4, 392 contamination by fungal parasites 399–400 712 Haematococcus pluvialis (Continued ) genetic improvement prospects 394 mass cultivation 396–9, 398, 480, 683 morphology and reproduction 389, 389–90 optimum growth and productivity conditions 396, 683 stress response physiology of protective mechanism 395, 395 pigment and structural changes 390, 390–1 haemolytic poisoning 587 half-saturation constants (nutrient uptake) 129–30 ‘Hanging Gardens’ vertical PBR design 236 Hansen solubility parameters 285–6 haptophytes 15 harmful algal blooms (HAB) algal species involved 584, 585–6, 672 increase in frequency 582, 582–3 prediction and control 589–90 harvesting cell aggregation by coagulation/flocculation 268–71, 270, 271, 272 centrifugal recovery 274–6, 287 draining of PBRs 452, 453 efficiency 351, 600 gravitational separation 271–3 method selection criteria 277–8, 301, 350–1, 656 non-chemical aggregation methods 276–7 pressure/vacuum filtration 273–4, 276, 287 regime, effect on cell density and productivity 199, 199 technical challenges 70, 267–8, 364 haslenes 521 hatcheries (farmed aquatic animals) algal culturing technologies 630–32, 630-33, 646 marine finfish production 618, 622, 636–7 types, requirements for microalgae 616–17 headspace volume effect on hydrogen photoproduction 424–6 in hybrid PBRs 242, 243, 244 health benefits of β-carotene 364–5 claimed benefits of Spirulina 353, 354, 475, 489, 490 effects of chlorophyll consumption 477-8, 505–6 hazards, algal toxins 482–3 pathogen destruction in waste stabilization ponds 596–7, 598 Index regulations for product quality and safety 353, 664 safety of operators in toxic algae culture 249 toxicological studies on algae as food 485–6 uses of antioxidants 388 heat tolerance 660–61 heavy metals collection of algal strains for bioremediation 73, 75, 606 concentrations in algae 483, 483 definition 602 detoxification methods 605–7 microalgal resistance mechanisms 604, 604–5 helical photobioreactors (biocoils) 234, 632 herbicide resistance, transgenic 658–9 heterokont algae, major clades 14–15 heterologous elements in genetic manipulation 147–8, 155, 156, 524 heterotrophic production advantages and commercial success 134, 257, 445, 686 heterotrophic–phototrophic sequential culture 119 potential new commercial products 139–43 see also fermentation heterotrophy capacity for, in microalgae 135–6, 136, 346, 399 collection of heterotrophic microalgae 72 definition and types 123–4, 124 HIV infection 491–2, 508 homogenizers, for cell disruption 282–3 homologous recombination 151–2, 153, 540 HPLC (high-performance liquid chromatography), lipids analysis 60–1 human nutrition see food hybrid photobioreactor design systems 241–4, 243, 253, 256 hydrocarbons biosynthetic pathways 519–21, 520 in Botryococcus braunii 372–4, 373, 374, 521 end uses and algal sources 369, 370, 522–3, 523 removal from waste in oxidation ponds 599 hydrogen production algal metabolic processes 418–20, 419 efficiency and economic feasibility 428 net energy ratio assessment 427 theoretical, energy capture and losses 420, 420–1 as energy resource, alternative options 417–18 PBR design 231, 423–7, 425 sulfur-starvation protocol 422, 423 hydrogenase enzymes 419–20, 421 identification techniques 87–8, 340, 434–6 IGV Biotech/GmbH basic design of photobioreactor systems 448, 452, 452–3 case studies of industrial facilities 453–5, 454, 455, 456 recent innovations in design 456–9, 457, 458 scale of designed products 448, 451, 451, 453 technological development and patents timeline 449–50 illuminated surface productivity (ISP) 255, 313 illumination laboratory and industrial scale sources 452 light concentration and distribution devices 249–51, 250 light-emitting diode (LED) sources 251 optical fibres 447–8 sources and regimes for culture collections 83–4 two-side, in flat-plate bioreactors 208, 215–16, 217–18, 218 immobilized cultures for aqueous heavy metal removal 73, 606 support materials 48–9 Immulina 492 immunomodulatory algal products 489, 510, 510–11 implied warranty for fitness (products) 496 inclined (cascade) system cultivation 228, 228–9, 333 inducible gene control elements 148–9 infant formula, DHA as supplement 137–8, 318 influenza, algal antiviral agents 508, 508 inhibition of growth, in ultra-high-density cultures 182, 183–5, 184 introns, effects on gene expression 155 ion channel modulatory activity 508–10, 509, 584 ionic liquid extraction 290–1 iron biochemical effects of deficiency and excess 117 ocean fertilization 694, 696 irradiance average intensity calculation, in bioreactors 189–90 effect on photosynthetic rate 22, 22, 91, 91–4 energy available for hydrogen production 420, 420–1 measurement and units 23 Index penetration depth 176, 177 solar, typical levels 33–4, 190 isolation of biochemical product from biomass 284–6, 288–92 of microalgae from natural sources 37–8, 70–2 isopentenyl pyrophosphate (IPP) 372–3, 374, 391 Jerez (Spain) industrial facility 455, 455 Kennedy pathway 547–50, 548, 558 Kjeldahl analysis method 512 ă Klăotze, OPA industrial facility, (Germany) 454 lactoferricin 659 lag phase (growth) 43 land requirements (for production plant) 322, 363, 383–4 LED lighting for photobioreactors 251 legislative provisions dumping of pollutants at sea 699 equipment standards 452–3 food additives 479, 495 life cycle assessment (LCA) 382, 427, 567, 573–4 light colour, effect on Botryococcus braunii 376, 377 photon flux density (PFD) 94, 171–2 properties and measurement 22–3, 23 thermal dissipation mechanisms 30–1, 96–7 see also illumination light acclimation carbohydrate and lipid accumulation 115 changes in pigmentation 30, 94–6, 115 in continuous and flashing light 221–2 failure, in low-density cultures 174–5, 175 inactivation of PS II 30–1, 97–9 in turbulent mass cultures 189 understanding of photoprotective mechanisms 96–7, 106, 361, 567 light reactions (of photosynthesis) 22, 22, 25–9 light–dark (LD) cycles frequency and culture turbulence 180–1 effect on hydrogen output 423 related to cell density and productivity 208–9 with long and short optical paths 182, 188, 188–9 resulting from mutual shading 173 timescale related to cell motion 187–8, 206–7, 215–16, 218–20 limiting factors light, diurnal patterns in outdoor cultures 192–3 in mass culture technology 99, 660 nutrients in ocean food webs 693 physiological definition 90, 92 resource competition 130 for specific applications 127 linear growth phase 44, 44 lipases, in algal oil conversions 558 lipid bodies and astaxanthin accumulation 390 biogenesis 553–5, 555 composition and structure 552, 552–3, 553, 554 functions, in eukaryotes 555–7 lipids accumulation in stress conditions 569 algal sources, quantities and uses 471–5, 517–22, 676–80 biosynthetic pathways 545, 547–52, 548 diversity in algae 12, 53 effects of light intensity 115 extraction form dry, wet, and paste biomass 289–90 fluorescence-based imaging and determination 62–4 organic solvent extraction 284–5, 286, 288–9 separation and quantification of different types 55–61, 62 total lipids analysis, gravimetric methods 53–5, 57 liquid–liquid product extraction 284–6 long optical paths 188 Lowry protein assay method 52 LPAAT (lysophospatidic acid acyltransferase) 549 lumens (lm), definition 23 luminostat operation 567 lutein factors affecting content in algae 684–5 production in serpentine tubular PBR 231 sources 480, 682 lux (lm m−2 ), definition 23 luxury uptake (nutrients) 127, 128 lycopene 392, 393, 507 lyophilization (freeze-drying) 280–1, 467, 634, 686 lysine 662–3 MAAs (mycosporine-like amino acids) 438, 508 macronutrient fertilization 694, 695, 696 magnetic separation of algal cells 277 maintenance coefficient 43 maintenance of cultures active cultures, growth conditions 39, 82–4 713 cryopreservation 86–7 nutritional status monitoring 199–200, 350 online productivity monitoring 197–9, 199 record-keeping and quality control 86, 349 requirements for high-density culture 182–5 strain purity maintenance 84–6, 200–1 usefulness of extremophilic strains 71–2, 200, 362 manifold photobioreactors 233–4, 234, 252 Manila clams (Ruditapes philippinarum) 640, 686 marine waters acidification 694, 700 changing nutrient ratios 584 collection and screening of algae 71, 74–5 fertilization/nourishment 694, 695–9, 697 location for floating photobioreactors 244–6, 624–5, 625 ocean carbon cycle 693–4 markers, genetic antibiotic/herbicide resistance selection 149, 409, 657 for identification 88, 434–6 range, in algae 537, 539, 540 mass culture downstream processing of algae 278–84, 634-5 effects of cell density 173–7, 174 harvesting 267–8 mixing/stirring requirements 177–80, 178, 179 multistage cultivation strategies 119, 125, 251, 254, 396–9 outsourcing by aquaculture hatcheries 631–2, 634, 646 performance monitoring 197–201 process control parameters 446, 446 resource and production variables 311, 318 theoretical and realistic photosynthetic efficiency 33–4, 194–7 mass spectrometry (MS) lipids analysis 58–61, 59 role in new product discovery 513–14, 514 maturation ponds (waste treatment) 597 mechanical cell disruption methods 282–4 media (for microalgal cultures) for Botryococcus braunii 374–5, 375, 376–7, 379 daily replacement for high-density culture 182–5, 183 fermentation (heterotrophic) 135–6 for Haematococcus pluvialis 396, 397 nutrient requirements 124–5, 126, 128 714 media (for microalgal cultures) (Continued ) organic load and contamination by competitors 200–1 types and components 82–3, 127, 630 Medusa design, axenic PBR 248 meganuclease-based gene insertion 152, 153, 156 Megazyme starch assay method 51 membrane filtration 273–4, 274 membrane trafficking 556 membranes, photosynthetic see thylakoids Mercenaria clams 640–1 mercury-induced toxicity 493, 603 metabolic engineering 155–9, 157–8, 515–17, 557 metal salts, use as flocculants 269 metal speciation 602–3, 603 methionone 662–3 mevalonate pathway 372, 391, 392 microalgal-to-biodiesel process economic assessment 297–301, 302, 303, 574 Nannochloropsis gaditana culture technology 293–6, 294 transesterification and FAME recovery 296, 296–7 microbial mats (biosorbents) 606 microcystins 586, 589 microencapsulation (for feed) 686 microfiltration 273–4 micronutrients anthropogenic increase and eutrophication 584 ocean fertilization 694, 696 supply requirements 127–8, 603 microscopy for contaminant monitoring 200 for direct isolation of microalgae 38 fluorescence imaging 62–3 optical cell counting 39 quality control in culture collections 86, 87 milk, effects of Spirulina in cows’ diet 487–8 mineral nutrition see nutrients minimal media 82, 85 mitochondria, diversity in algae 10 mitosis cell cycle and recombination types 152 diversity in algae 10 mitten crabs (Eriocheir sinensis) 637 mixing cost–benefit analysis 180 effects of different equipment 423, 424 effects of stirring rate on productivity 177–9, 178, 350 objectives and precautions 179–80 in paddle-wheel raceway ponds 348–9, 363 Index time determination, hydrogen production 424, 425, 426 using wave motion 624, 656 mixotrophic growth in Arthrospira (Spirulina) 346 in Botryococcus braunii 378–9 carbon supply and pH control 126 contamination risks 333, 379 definition 124 in Haematococcus pluvialis 399 modular reactor systems 119, 251, 448 moisture content 41 molluscs food sources for larvae and adults 618, 619, 639–41, 685 world production trends 617, 618, 618, 619 monoclonal antibodies 533 monounsaturated fatty acids (MUFAs) 517–18 morphology Arthrospira (Spirulina) 340–1, 341 Chlorella spp 330 colonial forms in Nostoc 433, 434, 435, 436 diversity in microalgae 3–8, 4, 5–6, Dunaliella salina 359–60, 360 Haematococcus pluvialis 389, 389–90 mRNA, expression control elements 147–9, 148 mud crabs (Scylla spp.) 637 mutagenesis insertional, for gene characterization 151 modification of Dunaliella for new products 364 for mutants with increased lipid production 524, 569 for promotion of homologous recombination 152 random, induced by UV/chemical treatment 160 MUTL (mesh ultra-thin layer) bioreactor design 456–7, 458, 459, 459 Nannochloropsis commercially produced for aquaculture 633, 633, 635 N gaditana, in microalgal-to-biodiesel process 293–6 Natural Products Association, US (NPA) 495 net energy ratio (NER) 427 net present value (NPV) 317, 318, 383, 383 net protein utilization (NPU) 468, 469 neurotoxic shellfish poisoning (NSP) 586 Neustadt-Glewe (Germany) industrial facility 454 neutral lipids 552 NHTR (Near-Horizontal Tubular Reactor) design 233, 241 night biomass loss (NBL) 198–9, 228 Nile Red (stain) 63, 63–4 nitrate reductase (NR) control elements 147, 148 nitrogen increase, in worldwide aquatic systems 584 non-protein, in microalgae 51–2 nutrient status, effects on algal biochemistry 116–17, 126–7 Botryococcus braunii 377–8 Chlorella spp 331 lipid content 473, 547, 569 ocean surface distribution, global 693, 693, 696 supply sources for microalgal culture 127, 135, 345 for ocean fertilization 697 suppression of nitrate use (transgenic mitigation) 661 uptake kinetics 128–30 nitrogen fixation (cyanobacteria) 72, 127, 693 nitrous oxide 701 nomenclature Arthrospira (Spirulina) taxonomic status 339–40, 463 symbols and units 303 non-photochemical quenching (NPQ) 32, 33, 94, 96 nonbiogenic toxins 482, 483 Nostoc growth and physiology 438–9 life cycle and colonial forms 434, 435, 436 mass cultivation 439–41, 440 nutritional and pharmaceutical value 436–8, 437, 492 species identification with molecular markers 434–6 traditional food use 433–4 NRPS (nonribosomal peptide synthetase) pathways 515, 516 nuclei, morphology and mitosis 10 nucleic acids, toxicity 482 nutrients in culture medium recipes 124–5, 126, 128 effect on toxicity of algae 576 effects of supply on microalgal composition 116–17, 523 imbalance and growth inhibition at high cell density 184–5 injection systems, for ocean fertilization 697–8 load, and eutrophication 581 monitoring in continuous cultures 199–200 natural resources availability 568–9 optimum ratios 130–1, 131 Index requirements of different trophic modes 123–4, 124 Arthrospira (Spirulina) 345–6 Botryococcus braunii 377–9 Chlorella 330–1 uptake kinetics 128–30, 129 nutritional supplements algae in animal feed 486–8, 685 edible vegetable oils (PUFAs) 137–8 evidence of toxicity 482–3, 486 market development 505 polysaccharides from red microalgae 412 products from Chlorella 137, 334 quality and safety of Nostoc 436–7, 437 with small-scale rural production technology 354 nutritional value, algal protein 463–8, 466 ocean biological carbon pump (OBCP) 693–4, 697 oceans see marine waters omega-3 fatty acids bio-rational collection of strains for production 75, 76, 139 early commercial fermentation production 136–7 enrichment by genetic engineering 663–4, 664 importance in aquaculture feeds 654 importance in aquaculture feeds 623, 629 nutritionally important, from algae 473, 475, 517 OMEGA (Offshore Membrane Enclosures for Growing Algae) floating PBRs 244–6, 245, 624 one-step absorption photosynthetic model 210, 210–11, 212 open cultivation systems (pond/raceway) for Arthrospira (Spirulina) 347–9, 348 for Chlorella spp 331, 333 compared with closed reactor systems 440–1, 445–6, 655–6 large-scale extensive production 362, 362–3 technologies 227–9, 228, 252, 256 used in aquaculture hatcheries 631, 631 operating costs (OPEX) aquaculture hatcheries 631 biodiesel case study 298, 299 oil-from-Botryococcus braunii case study 382–3 in techno-economic analysis 317, 318, 320, 321 optical density (OD) as biomass determination method 40, 40 monitoring for process control 453 optical path (OP) and areal cell density 185, 185, 185–7, 186 effect of length on productivity 181, 181, 181–2, 208, 410 related to cell motion and reaction timescales 187–9, 208 optimum culture conditions criteria for definition 84 light and temperature interaction 172, 344 N:P ratio 130–1, 131 optimal cell density (OCD) 173–7, 199, 208–9 strain specificity 375 organic solvent extraction of cell contents (cell disruption method) 282 enthalpy and solubility parameters 285–6 extraction of lipid from microalgae 284–5, 286, 288–90 ideal solvent characteristics 286 ornamental fish, colour enhancement 684 osmoregulation 10, 104–6, 118, 344 outdoor cultivation compared with heterotrophic fermentation 135 daily variation of irradiance and productivity 190–3, 191 open systems (pond/raceway) 227–9, 228, 252 photoacclimation and photoinhibition 96–7, 99–104, 174–5 photosynthetic efficiency 33–4, 106–7, 256 requirements for Arthrospira (Spirulina) culture 347 technologies for Botryococcus braunii 379–80, 380 output rate calculation, continuous flow culture 46, 46–7, 172 effect of population density 174, 174–6, 176 effect of reducing optical path 186, 187 interaction of light, OCD, and stirring speed 178–9, 179 oxidation ponds 597–601 oxidative stress bleaching damage threshold, Chlorella 330 pigment composition changes 391 protective physiology 395, 395, 478 oxygen control in closed PBR design 457–9 control in fermentation broths 135–6 effects of concentration on productivity 101–3, 103, 103, 570 in Arthrospira (Spirulina) 344–5 measurement of evolution (photosynthetic) 31, 198 ocean concentration 701 sensitivity of hydrogenase enzymes 421, 422 715 in waste treatment ponds, supplied by algae 596 oxylipins 518, 519 oysters (Crassostrea) aquaculture production systems 640 food requirements 618, 640, 641, 674, 680 greening, for increased market value 622 packaging of products 351–2 palmelloid growth form 5–6, PAP (phospatidic acid phosphatase) 549 Parallel Film Reactor (PFR) design 241 paralytic shellfish poisoning (PSP) 582-3, 583, 582 Paraphysoderma sedebokerensis (chytrid) 399–400 particle bombardment (biolistic) transformation 149–50, 151, 394, 536 partition coefficient, for solvent extraction 284 pastes, algal (aquaculture feed) 634, 685 patellamides 515, 517 pathogens, in domestic wastewater 598 PDAT (phospholipid:diacylglycerol acyltransferase) 550–1, 555 penaeids see shrimps peptide secondary metabolites 515 pesticides, isolation of tolerant strains 73 pH control for auto-flocculation 271, 272 balance in wastewater treatment ponds –597-598 carbonate/bicarbonate buffering for Arthrospira 345, 349–50 for elimination of contaminants 635 in mass culture 125–6, 127, 200 phenol–sulfuric acid method (total carbohydrates) 49–50 pheophorbides 477 phosphorus effects of depletion 117 requirements of Botryococcus braunii 378 supply and uptake 127, 128–30, 345–6 world resources and demand 568–9 phosphorylation ATP synthesis 26, 28–9 incorporation of orthophosphates 117 photic volume (PBRs) 173, 188, 208 photic zone (oceans) 694–5 photoacclimation see light acclimation photobioreactors (PBRs) commercial potential and economic factors 226–7, 256–7, 459, 459 compared with open pond systems 446, 655–6 comparison of types 205–6, 251–5, 252–4, 256 716 photobioreactors (PBRs) (Continued ) construction materials 427, 447, 568 definition, and category classifications 229 designed for light dilution 104 improved mixing rate 180–1 vertical arrangement 195, 426, 567, 567 designs for mass cultivation axenic and sensitive cultures 247–9 flat, rigid panel and flexible film 234–41, 235, 238, 240, 457 floating systems 244–6, 245, 624–5, 625 hybrid systems 241–4, 243 for substrate-attached organisms 246–7 tubular 230–4, 232, 234, 448 vertical columns and sleeves 229–30, 230 illumination systems 249–51, 250, 447–8 integrated, for multiple phase cultivation 119, 251, 364, 398 laboratory-scale apparatus 422–3, 448 productivity and efficiency assessment 195–7, 197, 255 risks of inadequate mixing 180 supplied to aquaculture hatcheries 632, 632 photoinhibition occurrence and recovery in Arthrospira 343 principles 22, 22, 97–103, 98 susceptibility of Nostoc colonies 439 photons, energy of 23, 33 photorespiration effects of oxygen concentration 102, 345 role of RuBisCO 30 photosynthesis crosstalk with carotenogenesis 394 dark reactions 29, 29–30 hydrogen generation 418–19, 419 light reactions 25–9, 27 monitoring techniques 31–2, 198 osmotic (salinity) stress responses 104–6, 344 process summary 21–2, 22 rate, light response (P/I) curve 22, 22, 91, 91–4, 172–3 role in environmental change signalling 91 timescale of reactions 187, 207, 209–12 photosynthetic efficiency differences between strains, Arthrospira 343 improvement, biological and technical approaches 566–7, 660 light conversion efficiency in H2 production 420–1, 428 microalgae, theoretical and actual 32–4, 103–4, 194–5 Index parameters and models 92–3 related to productivity 195, 256 photosynthetic units (PSU) 95, 95–6 photosynthetically active radiation (PAR) 23, 23, 171–2 photosystems I and II adaptations to salt stress 104–6 measurement of PS II quantum yield 93–4 models of photon absorption and recovery 209–12, 210, 211 PS II inactivation (in light acclimation) 30–1, 97–9 reaction centres 25–8, 26, 28 phycobilins/phycobiliproteins bioactive properties 438, 482, 492, 493 chemical structure 24, 25, 507 as fluorescent markers 482, 507 types and sources 481–2 phycobilisomes degradation in nitrogen deficiency 116 response to light conditions 96 structure and composition 26 physiology Arthrospira (Spirulina) 343–6 nutrition 17 osmotic regulation 10, 104–6, 118 responses to environmental stress 90–1, 107, 395–6 storage of photosynthetic products 9–10 use of genetic manipulation in research on 160 see also light acclimation; photosynthesis phytases 664 Phytobag reactor design 240, 241 phytoene synthase (PSY) 391–2 phytoplankton competition in growth media 200 oceanic abundance 13, 16–17, 615–16, 693 population responses to nutrient addition 701 response to eutrophication 582 picobiliphytes 16 pigments bioactive properties and uses 477–82, 505–7 effects of oxidative stress 391 effects of temperature increase 116 quantity measurement 25 types and roles in algae 23–5, 24, 26 pigs, algae as feed for 488 pipeline injection of ammonia (oceans) 697–8 PKS (polyketide synthase) pathways 136, 515, 516, 518 plastoquinone 27–8, 96, 361 poisoning incidents, from harmful algal blooms 582-583 see also toxins pollution bioremediation methods 605–7 dumping at sea 699 excess nutrient inputs 582, 645 microalgal biosensors 603 organic load of wastewater 597 toxic micropollutants 600, 602–3 polyelectrolytes 269–70, 270, 271 polyglucan granules 342 polyhedral bodies (carboxysomes) 342 polyhydroxyalkanoates (PHAs) 524, 524–5 polylactic acid (PLA) 524, 524 polyols, osmotic functions 118 polyphosphate granules, as storage bodies 117, 127 polyunsaturated fatty acids (PUFAs) from algae, quantities and uses 471, 473, 475, 517–18 chemical structure 517 effects of light intensity 115 enhancement in feed algae 663–4, 676–7 purification methods 293 world supply and demand 623–4 polyurethane foams (algal solid supports) 49 population density see cell concentration Porphyridium spp 406–12 poultry, algae as feed 486–7 pregnancy, effects of algal dietary supplements 490 preservation see maintenance of cultures preserved algae, for aquaculture feed 634, 685–7 pressurized liquid extraction 290 primary production, in aquatic food chain 616, 693–4, 695 probiotic algae 622, 636, 654, 687 process control systems 446, 446, 448, 452 process flow diagrams (PFD) 312, 312, 380 productivity consequences of photoinhibition 98, 98–104, 193 enhancement, future technological prospects 34, 226, 5557–558 improvement needed for commercial success 70, 566 oceans, stoichiometry 694–5 optimal cell density and light intensity 175, 175–6 reactor efficiency assessment 195–7, 255–6 theoretical limits, microalgal biomass 32–4 see also biomass profit, definition and calculation 311, 316, 317 prokaryotes evolutionary origins 12–13 structure and biochemistry 13, 14 see also bacteria; cyanobacteria Index promoters, transgenic expression 536–7, 538, 656–7 protein efficiency ratio (PER) 465–7, 466 proteins algal resources 12, 407–8, 674–6 amino acid composition 468–71, 470, 663, 676 associated with photosynthetic pigments 26–9, 28 iron-stress-induced 117 metal-binding, in detoxification 604 nutritional value parameters 463–8, 466, 469 produced by genetic manipulation 156–9, 157–8, 409–10, 532–40 quantitative measurement 51–3, 53, 465 structural, in lipid bodies 553 therapeutic, delivery systems to fish 665 ProviAPT plastic bag PBR design 239, 240 pseudopterosins 521–2 purification techniques axenic cultures 38 for extracted products 292–3 field samples 84–5, 440 pyrenoids 330 quality control of products, regulatory measures 352–3, 493–6, 494 protocols for culture collections 86 quantum yield (of photosynthesis) 92, 93–4, 99, 420, 420–1 quorum sensing 658, 660 quota flexibility (nutrients) 129, 129–30 rabbits, algae as feed 488 raceway ponds cost-effectiveness 256–7, 294–6, 295 coupled dual-cultivation systems with PBRs 251 covered hybrid systems 241, 244 open systems 227–8, 348–9 rate equations and constants, photosynthesis 210–12, 221 reactive oxygen species (ROS) effects on PS II damage and repair 97, 101–2 poisoning hazards 587, 603–4 related to oxidative stress 117, 395, 395 reactor volume efficiency 195–7, 197 recombinant proteins codon usage 154–5, 536–7, 537 expression promoters 536–7, 538 mammalian/bacterial platforms, and algal 532–3 products and sources 533, 534–5 potential of red microalgae 412 production from Chlorella 335–6 record-keeping, culture collections 86 recovery of algal cells see harvesting red algae 13, 14, 406–7 ‘red tides’ see algal blooms Redfield C:N:P ratio 131, 584, 694–5 refugee proteins 556 regulations food/dietary supplement quality and safety 353, 482, 493–6 transgenic approval, costs 655, 665 on uses of gray water 657–8 relative energy difference (RED), solvent extraction 286 reporter proteins 540 reservoirs, wastewater treatment 599 resistant starch 50 resolution (chromatography) 292–3 retinol (vitamin A) 364, 485, 490, 506, 681 revenues, estimation 318–19, 320 Reynolds number (turbulent flow) 180, 426 rheology, Porphyridium polysaccharides 408 riboflavin (vitamin B2 ) 680 risks, ocean nourishment 700-1 Ritschenhausen (Germany) industrial facility 454, 454–5 RNA interference (RNAi) gene knockdown 154 Rotating Algal Biofilm Reactor (RABR) design 246–7 rotifers mass production methods 643–4 microalgal requirements 622, 632 species used as food for fish-rearing 621, 643 RuBisCO enzyme 12, 29, 30, 457–8 ruminant animals, algae as feed 487–8 salinity and antibiotic effectiveness 149 collection of tolerant strains 74, 376 effect on microalgal composition 118, 360–1, 361 salt stress response, photosynthetic 104–6, 344 variation and Dunaliella biomass production 362–3, 363 salmon, methods of flesh pinkening 623 saturating pulse fluorescence 100 saturation irradiance (Ik ) 92, 93, 172, 343 saxitoxin 584, 588 scale-up to commercial production anticipated timescale 566 difficulties for Arthrospira (Spirulina) culture 349 economic aspects 298, 298, 459, 459 technological development for 226, 451, 571 717 scallops (Pecten), food requirements 618 Schizochytrium, fermentation processes biodiesel production 142 DHA and EPA production 138–9, 140, 142 screening direct and indirect assays 39, 513, 514 high-throughput techniques 75, 77 lipid content, by fluorescence spectroscopy 63–4 for multiple character traits 73–5, 76 for specific properties 70, 71, 72–3 sea bream, feed requirements 622, 637, 686 sea cucumber aquaculture 642–3, 646 seasonal cycles limiting mass culture production 347, 363 natural algal succession 17 product quality 352–3, 353 sunlight intensity variation 190 in wastewater treatment ponds 600 secondary carotenoids biosynthetic pathways 393 photoprotective role 115, 395 production in stress conditions 25, 116–17 secondary metabolites, bioactivity 438, 507–12 sedimentation, for cell recovery 271–3, 272 selection markers antibiotics/herbicide resistance 149, 409, 657 mutant complementation 149, 537 range available in algae 537, 539, 540 visible phenotypic, and fluorescence labelling 160 semi-continuous culture, rotifers 644 sequential collection photosynthetic model 209–10, 218–20 sequestration efficiency 697, 698 serpentine photobioreactors 230–3, 232, 252 settling velocity in centrifuges, force calculation 275 gravitational 272 sewage, treated, for Botryococcus braunii cultivation 379 sexual reproduction capability in microalgae 8, 360, 390 induced, for breeding and selection 159–60 Shandong Province (China), aquaculture 643, 645–7 shelf life of products 351, 634, 686 shellfish see molluscs shellfish-poisoning toxins 584, 586 ship-based ocean urea injection 698, 699 short optical paths 188–9, 215–16 shrimps hatchery systems, microalgal needs 619–21, 621, 638–9 world production trends 618, 619 718 Sigma starch assay method 50–1 silicon, nutritional requirement 128, 695 SimgaeTM hybrid PBR design 242 skin care products, anti-inflammatory 412 sleeve reactors 229, 237, 411, 411, 630 soda lakes, phytoplankton ecology 342–3 software for modelling analysis economic 312, 320, 324 fluid dynamics behaviour 423 soil biphasic soil–water culture media 83 isolation of microalgae from 38, 73 solar drying, algal biomass 279, 279–80 solubility parameters 285–6 sonication see ultrasound Soxhlet extraction method (lipids) 54–5, 57, 288 sparging 126, 248, 568 species, definition and identification 81–2, 434–6 specific growth rate (μ) 42, 46, 128–30, 129, 172 spirolides 586, 586 Spirulina see Arthrospira spray drying, algal biomass 278–9, 685 squalenes, biosynthetic pathways 372–4, 374 stabilization ponds, waste treatment 596, 596–8, 597 starch conversion to triacylglycerol 551–2, 557 quantitative measurement 50–1 static cultures 84 stationary growth phase 44, 45, 184 storage of cultures (long-term) 39, 86–7 strains characterization by products, Botryococcus 370, 371–2 collection and isolation strategies 70–7, 76 identification and authentication 87–8, 340 improvement using biotechnology 569–71 maintenance and storage 39, 84–6 numbering designation in collections 82 selection criteria, Arthrospira (Spirulina) 349 stress response, definition 90–1 subcritical water extraction 290 sugars in Arthrospira trophic regimes 346 in B braunii mixotrophic growth 378 as carbon source for fermentation 135, 136, 142 in microalgal cell wall polysaccharides 407, 662, 663 sulfated polysaccharides biosynthesis in cell wall formation 408–9 characteristics and natural function 407–8 Index formation and environmental conditions 408 product properties and applications 408, 411–12, 437–8 from red microalgae mass cultivation 410–11, 411 sulfur deprivation, and hydrogen production 419, 422 sun drying, algal slurry 279 sunlight energy and surface intensity 190 exposure and shading, effect of cell density 174–5, 175 stabilization pond disinfection 598 variation in solar angle 193–4 supercritical fluid extraction 291–2, 293, 400 swimming crabs (Portunus spp.) 637 switchable polarity solvents 291 symbols and units, listed 303 tandem mass spectrometry (MS/MS) 60, 61 taxonomy bases for classification, Chlorella 330 major phylogenetic groups of algae 13–16 phylogenetic tree of Botryococcus 370–2, 371 species and strains 81–2 status of Arthrospira/Spirulina 339–40 techno-economic analysis (TEA) conceptual framework and process flow analysis 311–12, 312 data sources 318–19, 319 inputs and analysis 319–20, 321 microalgal-to-biodiesel case study 297–301, 299, 302 model assumptions and units 313, 314–15, 316 oil from Botryococcus braunii case study 380–4, 381, 382, 383 output metrics 316, 317, 318 sample TEA construction, Spirulina biomass 321–4, 323–4 spreadsheet software 312 technological development barriers to commercial success 69–70 control of cell composition 118–19 current economic viability of PBR designs 257, 459 ethical and societal issues 699-700 range of designs and systems, IGV GmbH 446–53, 449–50, 451 reasons for promising future prospects 354, 656 scale-up to commercial production 226, 298, 324–5, 451, 5751 temperature effects on lipid extraction 55, 56 effects on microalgal chemical composition 115–16 effects on nutrient uptake 131 isolation of heat-resistant strains 73–4, 660–61 low, effect on photoinhibition 100–1, 103 low-temperature preservation, algal pastes 634, 685 range for different Botryococcus braunii strains 375–6 requirements for Arthrospira (Spirulina) 344 terpene-based products 521–2 therapeutic products algal lipids 473, 475 algal proteins, scope for metabolic engineering 156–9, 157–8, 335, 533 animal testing 484–5, 491, 492–3 antioxidants 388, 437, 478–9 bioassay screening for 513–14 growth regulators and vaccines 665 properties of red microalgal polysaccharides 412 secondary algal metabolites 508–12 traditional herbal remedies using algae 436, 489 thiamine (vitamin B1 ) 681 thin-layer chromatography (TLC), lipids analysis 55–6, 57 thraustochytrids heterotrophic cultivation 134, 142 for oils production 72, 518 see also Schizochytrium thylakoids arrangement of reaction complexes 25, 26 effects of low temperature on lipid saturation 115 in pro- and eukaryotes 8–9, 9, 25, 341 tilapia farming 687–8 tilt angle, reactor surface 193–4, 194 timescales in photosynthetic process 187, 207, 209–12 in reactor light regimes (LD cycles) 187–8, 206–7, 215–16 synchronization under flashing light regimes 213–15, 214, 215, 216 total organic carbon (TOC) 42, 114, 198 toxic algae influence of nutrients on toxicity 588 PBR design for safe cultivation 247–9 potential for high-density mass cultivation 590 types of poisoning and damage 583–4 toxicological studies, animal and human 484–6 toxins biogenic and nonbiogenic types in algae 482–3 haemolysins 587 heavy metals 483, 483, 602–4 in industrial wastewater 598, 605–7 Index produced by cyanobacteria 586–7 shellfish-poisoning neurotoxins 586 toxicity testing, traditional and new algal foods 437, 482 trace elements see micronutrients transesterification enzyme-based techniques 558 operational costs 300–1, 300–1, 302 process, in biodiesel production 296, 296–7 transformation choice of algae for development 658 gene targeting and knockdowns 151–4 methods 149–51, 150, 533, 536, 656 nuclear- and chloroplast-based 155, 533 stable, robust systems development 335, 409–10, 524, 657 transgenic microalgae advantages over transgenic terrestrial plants 156 bioactivity and uses 533 epigenetic gene silencing 154 inability to survive in the wild 661–62 level of expression, control 147–9 regulatory approval and acceptance 655 trait mining within a genus 159 trehalose, in osmoregulation 344 triacylglycerols (TAGs) biosynthesis 545, 547–51, 548, 558 detection of trace amounts 61 enhancement by metabolic engineering 557, 570 interaction with starch synthesis 551–2, 557 molecular fingerprinting 60 physiological functions 556–7 sequestration (lipid body biogenesis) 552–6, 554 trichomes 341, 351, 440 trimethylamine 665 trophic independence 623 tubular photobioreactors for Chlorella cultivation 333 designs 230–4, 232, 234, 448, 455 floating reactors 244, 245, 245–6 inflated bag hybrid designs 241–2, 243 performance assessment 293, 294 semicontinuous B braunii cultivation 379–80 turbidostat (constant volume) culture 47–8, 48 turbulence bubble-induced, empirical description 221 effect on motion of cells 187–8 energy input requirement 568 influence on light–dark cycles 180–1 ultrafiltration 273, 274, 274 ultrahigh cell density (UHCD) cultures 182–7, 205–9, 206 ultrasound cell agglomeration method 277, 277 for microalgal cell disruption 283–4 ultrastructure Arthrospira (Spirulina) 341–2 biogenesis of lipid bodies 552, 553–5, 556 changes during carotenogenesis 390, 390–1 Chlorella spp 330 effects of cell density 177 fluorescence imaging techniques 62–3 microalgal cells 8–11, unialgal cultures 85–6, 200, 350 units and symbols conversion factors 313, 314–15, 316 listed 303 UTEX 2629 strain, taxonomy 370–1 UV (ultraviolet) radiation effect of UV-B on nitrogen uptake 131 effects on Arthrospira (Spirulina) 341, 343–4 irradiation, for culture purification 38 protective agents, algal products 438 V-trough hybrid PBR designs 242 vacuoles, functions in microalgae 10 vertical alveolar panels (VAP) 193, 234–5 vertical column (cylinder) cultivation 229–30, 230, 252, 630 Vertigro film panel reactors 238–9 very long-chain fatty acids (VLCFAs) 372, 373, 547 viability, assessment using TEA 310–11, 324–5 viruses, potential control of 659–60 viscosity in high-density cultures 221, 268, 410 red microalgal polysaccharides 407, 408 vitamin B12 (cobalamin), bioavailability 475–7, 632 vitamins microalgal content 475–7, 476, 680–81, 681 requirements of Botryococcus braunii 375 volume consideration in choice of harvesting method 277 of containers for culture collections 84 volumetric productivity for evaluation of mass culture performance 255, 313 with long optical path 188 719 parameters affecting 197, 216–17, 217 related to culture density 207–8, 212 wastewater treatment with Botryococcus cultivation, cost benefits 379 collection/isolation of algal strains 72–3 heavy metal sequestration 606 industrial wastewater 598–9 potential of Chlorella 335 regulatory barriers to use for algaculture 657–8 stabilization ponds 596, 596–8, 597 water algal moisture content measurement 41, 463 aqueous extraction processing 290 as base substrate for cultures 82–3, 128 concentration of algae from samples 38 fresh, world supply and demand 569, 574 recirculation for biofiltering, mariculture 645 use in solvent extraction of lipids 281, 288 wet biomass measurement 40–1 www (‘windy, wavy, and wiped’) PBR design 231–2, 232, 624 xanthophyll cycle, excess energy dissipation 96, 98 xanthophylls chemical structure 24, 478 occurrence in algae 25, 681 yessotoxins (YTX) 587 yield algaculture compared with agriculture 655 areal and volume, in reactors 195–6, 196 fish, correlation with phytoplankton 696 of microalgae in harvesting operations 268 of specific products and biomass 119 extracted by different methods 288–9, 289 substrate to biomass conversion efficiency 42, 346 see also output rate Z scheme (photosystem electron transport) 25–6, 27, 194–5 zeaxanthin 481 zooplankton contaminants, control 201, 659 production for aquaculture 643–4, 674 ... should be sought Library of Congress Cataloging-in-Publication Data Handbook of microalgal culture : applied phycology and biotechnology / edited by Amos Richmond and Qiang Hu – Second edition pages.. .Handbook of Microalgal Culture Applied Phycology and Biotechnology Second Edition Edited by Amos Richmond, Ph.D., Prof Emeritus Ben Gurion University of the Negev at Sede-Boker,... benefit of humanity Amos Richmond, PhD Prof Emeritus, Ben-Gurion University Sede-Boker, Israel Part The Microalgal Cell with Reference to Mass Cultures Handbook of Microalgal Culture: Applied Phycology