Marine Algae Extracts

Edited by Se-Kwon Kim and Katarzyna Chojnacka Marine Algae Extracts

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Edited by Se-Kwon Kim and Katarzyna Chojnacka

Marine Algae Extracts Processes, Products, and Applications

Volume 1

Edited by Se-Kwon Kim and Katarzyna Chojnacka

Marine Algae Extracts Processes, Products, and Applications

Volume 2

The Editors Prof. Se-Kwon Kim

Pukyong National University Marine Bioprocess Research Daeyeon-Dong, Nam-Gu 599-1 608-737 Busan South Korea

All books published by Wiley-VCH are carefully produced. Nevertheless, authors, editors, and publisher do not warrant the information contained in these books, including this book, to be free of errors. Readers are advised to keep in mind that statements, data, illustrations, procedural details or other items may inadvertently be inaccurate.

Katarzyna Chojnacka

Wroclaw University of Technology Institute of Inorganic Technology and Mineral Fertilizers Smoluchowskiego 25 50-373 Wroclaw Poland

Library of Congress Card No.: applied for

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Bibliographic information published by the Deutsche Nationalbibliothek

Poisonous Algae in the Red Sea. Source: Fotolia © irisphoto1

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V

Contents to Volume 1 List of Contributors XVII Preface XXVII Acknowledgments XXIX 1

Introduction of Marine Algae Extracts 1 Katarzyna Chojnacka and Se-Kwon Kim

1.1 1.2 1.3 1.4 1.4.1 1.4.2 1.4.3 1.4.4 1.4.5 1.4.6 1.5 1.6

Introduction 1 Algal Biomass as a Useful Resource 2 Biologically Active Compounds Extracted from Algae 4 The Application of Products Derived from Algal Biomass 5 Agriculture – for Plants 6 Functional Food 7 Cosmetics 7 Pharmaceuticals 8 Fuels 8 Antifouling Compounds 8 Extraction Technology 9 Conclusions 10 References 11 Part I: Cultivation and Identification of Marine Algae 15

2

Identification and Ecology of Macroalgae Species Existing in Poland 17 Beata Messyasz, Marta Pikosz, Grzegorz Schroeder, Bogusława Łe˛ska, and Joanna Fabrowska

2.1 2.2 2.3

Introduction 17 Collection of Macroalgal Thalli and Culture Conditions 20 Macroalgae Forming a Large Biomass in Inland Waters of Poland 21 Ecology Aspects of Freshwater Macroscopic Algae 31 Summary 33

2.4 2.5

VI

Contents

Acknowledgments References 34

34

3

Identification of Microalgae Producers of Commercially Important Compounds 41 Rosalia Contreras, J. Paniagua-Michel, and Jorge Olmos

3.1 3.2 3.2.1 3.2.2 3.2.3 3.3 3.4 3.5 3.5.1 3.5.2 3.5.2.1 3.5.2.2 3.5.2.3 3.5.2.4 3.5.2.5 3.5.2.6 3.5.3 3.6

Introduction 41 Microalgae for Human Consumption 41 Chlorella 43 Dunaliella 43 Haematococcus Pluvialis 44 Microalgae for Aquaculture and Animal Farms Microalgae for Biofuels 46 Molecular Identification of Microalgae 47 MA1-MA2 Universal Oligonucleotides 47 Amplification of the 18S rDNA Gene 48 Dunaliella 48 Botryococcus 50 Chlamydomonas 50 Scenedesmus 52 Chlorella 53 Other Microalgae Genera 53 18S rDNA Introns Characterization 53 Conclusion 54 References 55

4

Cultivation and Identification of Microalgae (Diatom) 59 Sekar Ashokkumar, Kuppusamy Manimaran, and Keun Kim

4.1 4.2 4.2.1 4.2.2 4.2.3 4.2.3.1 4.2.3.2 4.2.3.3 4.2.3.4 4.2.3.5 4.3 4.3.1 4.3.1.1 4.3.1.2 4.3.1.3 4.3.1.4 4.3.1.5

Introduction 59 Materials and Methods 61 Plankton Net 61 Preparation for Light Microscopy 62 Identification of Species 62 Odontella Mobiliensis (Bailey) Grunow 1884 62 Pleurosigma Normanii 63 Chaetoceros Curvisetus 64 Skeletonema Costatum 64 Coscinodiscus Centralis 65 Algal Culture Conditions 66 Physical and Chemical Conditions 66 Light 67 Temperature 67 Salinity 68 pH 68 Aeration/Mixing 68

45

Contents

4.3.1.6 4.3.2 4.3.3 4.3.4 4.3.5 4.3.5.1 4.3.5.2 4.3.5.3 4.3.5.4 4.3.5.5 4.3.6 4.3.7 4.3.7.1 4.4

Culture Medium/Nutrients 68 Isolating/Obtaining and Maintaining of Cultures Sources of Contamination and Treatment 70 Algal Culture Techniques 71 Growth Dynamics 71 Lag or Induction Phase 71 Exponential Phase 72 Phase of Declining Growth Rate 72 Stationary Phase 72 Death or Crash Phase 72 Harvesting and Preserving Microalgae 72 Algal Production Cost 72 Uses of Algae 73 Conclusion 73 References 73

69

Part II: Production and Processing of Marine Algae 79 5

Analysis of Green Algae Extracts 81 Grzegorz Schroeder, Bogusława Łe˛ska, Joanna Fabrowska, Beata Messyasz, and Marta Pikosz

5.1 5.2

Introduction 81 The Algae Biomass as a Raw Material of Natural Chemical Compounds 82 Methods of Extraction of Biochemical from Algae Biomass Analytical Procedures 87 Conclusion 92 Acknowledgments 93 References 93

5.3 5.4 5.5

6

Algae Extract Production Methods and Process Optimization Edward Rój, Agnieszka Dobrzyńska-Inger, Agnieszka De˛bczak, Dorota Kostrzewa, and Katarzyna Ste˛pnik

6.1 6.2 6.3

Introduction 101 Production Methods 102 Analytical Methods Used for Extract Production Process Control 108 Process Optimization 111 Example of Process Optimization 113 Materials and Methods 113 Experiments and Results 114 Summary 117 Acknowledgments 118 References 118

6.4 6.4.1 6.4.1.1 6.4.1.2 6.5

85

101

VII

VIII

Contents

7

Production of Seaweed Extracts by Biological and Chemical Methods 121 Izabela Michalak and Katarzyna Chojnacka

7.1 7.2 7.3 7.4 7.5 7.5.1 7.5.2 7.5.2.1 7.5.2.2 7.6

Introduction 121 Production of Algal Extracts with Different Methods 122 Pretreatment of Algal Biomass and Extraction Procedure 123 Algal Extracts Obtained by Enzymatic Hydrolysis 126 Algal Extracts Obtained by Chemical Hydrolysis 127 Extraction with Organic Solvents 127 Extraction with Inorganic Solvents 128 Acid and Alkali Hydrolysis 129 Extraction with Distilled Water 129 Comparison of Extraction Methods of Biologically Active Compounds from Seaweeds 130 Evaluation of the Activities of Algal Extracts Obtained by the Extraction with Organic Solvent 131 Antioxidant Properties of Seaweed Extracts 131 Antimicrobial Activity of Seaweed Extracts 132 Antiviral Activity of Seaweed Extracts 133 Anti-inflammatory Activity of Seaweed Extracts 133 The Application of Water Extracts from Seaweeds 133 Examples of Commercial Products Obtained by Extraction form Seaweeds 138 Conclusions 139 Acknowledgments 139 References 139

7.7 7.7.1 7.7.2 7.7.3 7.7.4 7.8 7.9 7.10

8

Upstream Processing in the Technology of Algal Extracts: Biomass Harvesting and Preparation for Extraction Process 145 Radoslaw Wilk and Katarzyna Chojnacka

8.1 8.1.1 8.1.2 8.2 8.2.1 8.2.2 8.2.3 8.2.4 8.2.5 8.3

Introduction 145 Cells Harvesting/Separation 147 Cells Disintegration and Extraction 149 Marine Vegetation from Baltic Sea as Source Material 149 Biomass Characterization 149 Legal Regulations 150 Availability of Marine Biomass in Poland 151 Biomass Collection Techniques 151 Method for Evaluating the Machines 152 The Technology of Raw Marine Biomass Preparation for Supercritical Fluid Extraction 152 Basic Operations 153 Algae Raw Material Treatment Methods 154 Acid’s and Alkalia’s 154 Pilot Plant Investigation and Cost Analysis 154

8.3.1 8.3.2 8.3.2.1 8.3.3

Contents

8.4

Conclusions 157 Acknowledgments 157 References 157

9

Downstream Processing in the Technology of Algal Extracts – From the Component to the Final Formulations 161 Radosław Wilk and Katarzyna Chojnacka

9.1 9.2 9.2.1 9.2.1.1 9.2.2

Introduction 161 Final Formulation 163 The Concept of Formulations 165 Adjuvants and Additives 165 Physical and Chemical Properties that Determine Effective Uptake of Active Ingredients Contained in Product 168 Solubility 168 Solution pH 168 Particle Size 168 Concentration of Active Ingredients 168 Common Formulation Types 168 Definition of an Emulsion 169 The Method to Produce an Emulsion Based on Algae Extract 170 Soluble Liquid (SL) 172 Emulsifiable Concentrate (EC) 173 Suspension Concentrate (SC) 173 Suspoemulsion (SE) 174 Seed Treatments (FSs) 174 Oil Dispersion (OD) 175 Stability of Algae Extract Emulsion 175 Conclusion 177 References 177

9.2.2.1 9.2.2.2 9.2.2.3 9.2.2.4 9.2.2.5 9.3 9.4 9.4.1 9.4.2 9.4.3 9.4.4 9.4.5 9.4.6 9.5 9.6

10

Algae Biomass as a Raw Material for Production of Algal Extracts 179 Agnieszka Saeid and Katarzyna Chojnacka

10.1 10.2 10.3 10.3.1 10.3.2 10.3.2.1 10.3.2.2 10.3.2.3 10.3.2.4 10.3.2.5 10.3.2.6 10.4

Introduction 179 Cell Wall 179 Methods of Obtaining the Biomass of Algae 181 Macroalgae 181 Microalgae 182 Open Raceway Ponds Versus Closed Photobioreactors Contamination 183 Productivity 184 Photosynthesis Conditions 185 Mixing 185 Feeding Strategies 185 Conclusions 186 References 187

183

IX

X

Contents

11

Algal Extracts as Plant Growth Biostimulants 189 Katarzyna Chojnacka, Izabela Michalak, Agnieszka Dmytryk, Mateusz Gramza, Adam Słowiński, and Henryk Górecki

11.1 11.2 11.3 11.4

Introduction 189 The Development of Fertilizers Industry 190 Plant Biostimulants 194 Potential Benefits Arising from the Use of Plant Growth Biostimulants 195 The Market of Biostimulants 196 Seaweed Biomass as a Source for the Production of Algae Based Fertilizers 197 Algae as the Resource for Biostimulants Production 199 Methods of Production of Commercial Biostimulants from Algae 201 Characteristics of Biostimulants Derived from Algae 202 Current Market of Algal Plant Growth Stimulants 204 Perspectives 205 Regulations 206 Conclusions 207 Acknowledgments 208 References 208

11.5 11.6 11.7 11.8 11.9 11.10 11.11 11.12 11.13

12

Effects of Alginate Oligosaccharides on the Growth of Marine Microalgae 213 Mikinori Ueno and Tatsuya Oda

12.1 12.2 12.3

Introduction 213 Preparation of Alginate Oligosaccharides 215 Effects of Alginate Oligosaccharides on the Growth of Nannochloropsis oculata 217 Species-Specific Effects of Alginate Oligosaccharides on the Growth of Diatom Chaetoceros gracilis and Skeletonema sp. 218 Effects of Alginate Oligosaccharides on Harmful Microalgae 220 Conclusion 222 References 222

12.4 12.5 12.6

Part III: Marine Algal Products 227 13

Omegas: Pharmaceutical High Value Products and One of the Most Functional Bioactive Compounds 229 Viviana P. Rubio, J. Paniagua-Michel, and Jorge Olmos

13.1 13.2 13.2.1 13.2.2 13.3

Introduction 229 Most Functional Omegas 231 Omega-3 231 Omega-6 232 Biosynthesis and Functions 232

Contents

13.4 13.5 13.6 13.7 13.8

Omegas and Diet 234 Omegas; Sickness; and Health 235 Omegas: Commercial Applications 236 Microalgae as a Source for Omega Production Perspectives 241 References 241

14

An Overview of Global Distribution of the Diterpenes Synthesized by the Red Algae Laurencia Complex (Ceramiales, Rhodomelaceae) 245 Luciana R. de Carvalho, Julyana N. Farias, Pablo Riul, and Mutue T. Fujii

14.1 14.2 14.3

Introduction 245 Biosynthesis of Diterpenes 246 Diversity and Geographic Distribution of the Diterpenes in Laurencia Complex 256 Conclusions 261 Acknowledgments 262 References 262

14.4

237

15

Anticancer Compounds from Marine Algae 267 Yong-Xin Li, Yong Li, and Se-Kwon Kim

15.1 15.2 15.3 15.4 15.5

Introduction 267 Terpenoids from Marine Algae 268 Sterols from Marine Algae 270 Polysaccharides from Marine Algae 273 Summary 274 Acknowledgments 274 References 274

16

A Comparative Analysis of Carrageenans Produced by Underutilized versus Industrially Utilized Macroalgae (Gigartinales, Rhodophyta) 277 Leonel Pereira, Filipa Meireles, Helena T. Abreu, and Paulo J.A. Ribeiro-Claro

16.1 16.1.1 16.1.2 16.1.3 16.1.4 16.2 16.3

Introduction 277 Phycocolloids from Red Algae 277 Carrageenan and Carrageenan Industry 278 Macroalgae Producing Carrageenan 279 Integrated Multitrophic Aquaculture (IMTA) 285 Chondrus crispus IMTA Cultivated 286 Geographic Localization, Date of Harvest, Yields, and Phycocolloid Type Produced by Red Algae 287 Analysis of Carrageenan by Vibrational Spectroscopy 287 Conclusion 288 Acknowledgments 290

16.4 16.5

XI

XII

Contents

List of Abbreviations and Symbols References 291

290

17

Biosynthesis of Nanoparticles Using Marine Algae: A Review 295 Panchanathan Manivasagan and Se-Kwon Kim

17.1 17.2 17.3 17.4 17.5 17.6

Introduction 295 Types of Nanoparticles 296 Characterization of Nanoparticles 297 Biosynthesis of Nanoparticles by Marine Algae Applications of Nanoparticles 301 Conclusions 302 Acknowledgments 302 References 302

18

Enzyme-Assisted Extraction to Prepare Bioactive Peptides from Microalgae 305 H.H. Chaminda Lakmal, Kalpa W. Samarakoon, and You-Jin Jeon

18.1 18.2 18.3 18.3.1 18.3.2 18.3.3 18.4 18.5

Introduction 305 Enzyme-Assisted Extraction and Isolation of Bioactive Peptides Bioactivity of Peptides Derived from Marine Microalgae 309 Antioxidant 309 Antihypertensive 311 Other Bioactivity 312 Molecular Modeling 312 Future Trends and Prospective 315 References 315

19

An Overview of Phycocolloids: The Principal Commercial Seaweed Extracts 319 Ratih Pangestuti and Se-Kwon Kim

19.1 19.2 19.3 19.3.1 19.3.2 19.3.3 19.4 19.4.1 19.4.2 19.4.3 19.5 19.5.1 19.5.2 19.5.3

Introduction 319 General Properties of Phycocolloids 320 Agar 320 Source and Extraction 321 Food Applications and Health-Promoting Effects 322 Technological Applications 322 Alginates 322 Source and Extraction 323 Food Application and Health Promoting Effects 323 Technological Applications 325 Carrageenan 325 Source and Extraction 326 Food Applications and Health-Promoting Effects 326 Technological Applications 328

298

306

Contents

19.6

Conclusions 329 References 329

20

Analytical Approaches for the Detailed Characterization of Microalgal Lipid Extracts for the Production of Biodiesel 331 Damien L. Callahan, Gregory J.O. Martin, David R.A. Hill, Ian L.D. Olmstead, and Daniel A. Dias

20.1 20.1.1

Introduction 331 Microalgal Lipids and Characteristics of Interest to Biodiesel Production 331 Effect of Fatty Acid Profile in Lipids on Biodiesel Quality 333 Characterization of Microalgal Lipids 333 Protocols 336 Lipid Extraction 336 Algae Culture 336 Protocol 1 Total Fatty Acid (TFA) 337 Protocol 2 Free Fatty Acid (FFA) and Polar Metabolites 337 Solid-Phase Extraction of Lipids (SPE) 337 Gas Chromatography–Mass Spectrometry (GC–MS) 339 Derivatization 340 Derivatization of Extracted Fatty Acids to Produce FAMEs 340 GC–MS Conditions 340 Liquid Chromatography/Liquid Chromatography–Mass Spectrometry 341 Liquid Chromatography 341 Liquid Chromatography–Mass Spectrometry 342 Combined Approaches 344 Final Remarks 344 Acknowledgments 344 References 345

20.1.2 20.1.3 20.2 20.2.1 20.2.2 20.2.3 20.2.4 20.3 20.4 20.5 20.5.1 20.5.2 20.6 20.6.1 20.6.2 20.7 20.8

Contents to Volume 2 List of Contributors XV Preface XXV Acknowledgments XXVII Part IV: Biological Applications of Marine Algae 347 21

Algal Extracts in Dentistry 349 Marcin Mikulewicz and Katarzyna Chojnacka

XIII

XIV

Contents

22

Marine Algae for Protecting Your Brain: Neuroprotective Potentials of Marine Algae 359 Pradeep Dewapriya and Se-Kwon Kim

23

Antiviral Activities of Marine Algal Extracts 371 Fatih Karadeniz, Mustafa Z. Karagozlu, and Se-Kwon Kim

24

Antihyperglycemic of Sargassum sp. Extract 381 Muhamad Firdaus, Rahmi Nurdiani, and Asep A. Prihanto

25

Immunological Activity of Marine Microalgae Extracts 395 Mariangela Caroprese, Maria G. Ciliberti, and Marzia Albenzio

26

Algal Polysaccharides and Their Biological Applications 413 Sirisha L. Vavilala and Jacinta S. D’Souza

27

Biological Phlorotannins of Eisenia bicyclis Sang-Hoon Lee and Se-Kwon Kim

453

Part V: Biomedical Applications of Marine Algae 465 28

Algal Extracts as a Carrier of Micronutrients – Utilitarian Properties of New Formulations 467 Łukasz Tuhy, Katarzyna Chojnacka, Izabela Michalak, and Anna Witek-Krowiak

29

Marine Algae Based Biomaterials for Osteoblast Differentiation and Tissue Regeneration 489 Pathum Chandika and Won-Kyo Jung

30

Marine Algae Derived Polysaccharides for Bone Tissue Regeneration 509 Jayachandran Venkatesan and Se-Kwon Kim

31

Wound Dressings from Algal Polymers 523 Monica Bhatnagar and Ashish Bhatnagar

32

Marine Algae and Chronic Diseases 557 Kalimuthu Senthilkumar and Se-Kwon Kim

33

Algae Wastes Biomass – a New Class of Low-Cost Material with Potential Applications in Environmental Engineering 575 Laura Bulgariu and Dumitru Bulgariu

Contents

Part VI: Food and Industrial Applications of Marine Algae 603 34

Algae Extract as a Potential Feed Additive 605 Mariusz Korczyński, Zuzanna Witkowska, Sebastian Opaliński, ´ Marita Swiniarska, and Zbigniew Dobrzański

35

Application of Marine Algae Derived Nutraceuticals in the Food Industry 627 Isuru Wijesekara and Se-Kwon Kim

36

Microalgal Carotenoids: Bioactive Roles, Health Foods, and Pharmaceuticals 639 J. Paniagua-Michel, Jorge Olmos Soto, and Eduardo Morales Guerrero

37

Biologically Active Organic Compounds, Especially Plant Promoters, in Algae Extracts and Their Potential Application in Plant Cultivation 659 Bogusława Górka, Jacek Lipok, and Piotr P. Wieczorek

38

Biomass and Extracts of Algae as Material for Cosmetics 681 Joanna Fabrowska, Bogusława Łe˛ska, Grzegorz Schroeder, Beata Messyasz, and Marta Pikosz Index

707

XV

V

Contents to Volume 1 List of Contributors XVII Preface XXVII Acknowledgments XXIX 1

Introduction of Marine Algae Extracts 1 Katarzyna Chojnacka and Se-Kwon Kim Part I: Cultivation and Identification of Marine Algae 15

2

Identification and Ecology of Macroalgae Species Existing in Poland 17 Beata Messyasz, Marta Pikosz, Grzegorz Schroeder, Bogusława Łe˛ska, and Joanna Fabrowska

3

Identification of Microalgae Producers of Commercial Importance Compounds 41 Rosalia Contreras, J. Paniagua Michel, and Jorge Olmos

4

Cultivation and Identification of Microalgae (Diatom) 59 Sekar Ashokkumar, Kuppusamy Manimaran, and Keun Kim Part II: Production and Processing of Marine Algae 79

5

Analysis of Green Algae Extracts 81 Grzegorz Schroeder, Bogusława Łe˛ska, Joanna Fabrowska, Beata Messyasz, and Marta Pikosz

6

Algae Extract Production Methods and Process Optimization Edward Rój, Agnieszka Dobrzyńska-Inger, Agnieszka De˛bczak, Dorota Kostrzewa, and Katarzyna Ste˛pnik

101

VI

Contents

7

Production of Seaweed Extracts by Biological and Chemical Methods 121 Izabela Michalak and Katarzyna Chojnacka

8

Upstream Processing in the Technology of Algal Extracts: Biomass Harvesting and Preparation for Extraction Process 145 Radoslaw Wilk and Katarzyna Chojnacka

9

Downstream Processing in the Technology of Algal Extracts – From the Component to the Final Formulations 161 Radosław Wilk and Katarzyna Chojnacka

10

Algae Biomass as a Raw Material for Production of Algal Extracts 179 Agnieszka Saeid and Katarzyna Chojnacka

11

Algal Extracts as Plant Growth Biostimulants 189 Katarzyna Chojnacka, Izabela Michalak, Agnieszka Dmytryk, Mateusz Gramza, Adam Słowiński, and Henryk Górecki

12

Effects of Alginate Oligosaccharides on the Growth of Marine Microalgae 213 Mikinori Ueno and Tatsuya Oda Part III: Marine Algal Products 227

13

Omegas: Pharmaceutical High Value Products and One of the Most Functional Bioactive Compounds 229 Viviana P. Rubio, J. Paniagua-Michel, and Jorge Olmos

14

An Overview of Global Distribution of the Diterpenes Synthesized by the Red Algae Laurencia Complex (Ceramiales, Rhodomelaceae) 245 Luciana R. de Carvalho, Julyana N. Farias, Pablo Riul, and Mutue T. Fujii

15

Anticancer Compounds from Marine Algae 267 Yong-Xin Li, Yong Li, and Se-Kwon Kim

16

A Comparative Analysis of Carrageenans Produced by Underutilized Versus Industrially Utilized Macroalgae (Gigartinales, Rhodophyta) 277 Leonel Pereira, Filipa Meireles, Helena T. Abreu, and Paulo J.A. Ribeiro-Claro

17

Biosynthesis of Nanoparticles Using Marine Algae: A Review 295 Panchanathan Manivasagan and Se-Kwon Kim

Contents

18

Enzyme-Assisted Extraction to Prepare Bioactive Peptides from Microalgae 305 H.H. Chaminda Lakmal, Kalpa W. Samarakoon, and You-Jin Jeon

19

An Overview of Phycocolloids: The Principal Commercial Seaweed Extracts 319 Ratih Pangestuti and Se-Kwon Kim

20

Analytical Approaches for the Detailed Characterization of Microalgal Lipids Extracts for the Production of Biodiesel 331 Damien L. Callahan, Gregory J.O. Martin, David R.A. Hill, Ian L.D. Olmstead, and Daniel A. Dias

Contents to Volume 2 List of Contributors XV Preface XXV Acknowledgments XXVII Part IV: Biological Applications of Marine Algae 347 21

Algal Extracts in Dentistry 349 Marcin Mikulewicz and Katarzyna Chojnacka

21.1 21.2

Introduction 349 Various Applications of Products Derived from Algae in Dentistry 349 Impression Materials 349 Agar and Alginate Hydrocolloid Impression Material 351 Other Hydrocolloids (Agar) 352 Toothpastes 352 Mouthwash 352 Anti-Inflammatory Applications 353 Alloplastic Synthetic Grafts (Fluorohydroxyapatitic Biomaterial) 353 Biocompatibility 354 Additional Applications 355 Potential Application of Mineralization Properties 355 Biomaterials 355 Antiplaque and Anticalculus Properties 355 Regenerative Materials in Periodontal Diseases 355 Chewing Gums 355 Conclusions 356 References 357

21.2.1 21.2.1.1 21.2.1.2 21.2.2 21.2.3 21.2.4 21.2.5 21.2.6 21.3 21.3.1 21.3.2 21.3.2.1 21.3.3 21.3.4 21.4

VII

VIII

Contents

22

Marine Algae for Protecting Your Brain: Neuroprotective Potentials of Marine Algae 359 Pradeep Dewapriya and Se-Kwon Kim

22.1 22.2

Introduction 359 Neuroprotective Properties of Algae and Algae-Derived Compounds 360 Anti-Inflammatory Compounds 360 Compounds against Oxidative Stress and Mitochondrial Dysfunction in Neuron 362 Marine Algae against Aggregated Misfolded Proteins-Induced Neurotoxicity 364 Cholinesterase Inhibitory Activity 365 Other Algae-Derived Neuroprotective Materials 366 Concluding Remarks 367 References 367

22.2.1 22.2.2 22.2.3 22.2.4 22.2.5 22.3

23

Antiviral Activities of Marine Algal Extracts 371 Fatih Karadeniz, Mustafa Z. Karagozlu, and Se-Kwon Kim

23.1 23.2 23.2.1 23.2.2 23.2.3 23.2.4 23.3

Introduction 371 Substances Responsible for Antiviral Activity of Algal Extracts Phlorotannins 372 Polysaccharides 374 Lectins 376 Others 377 Conclusion 377 References 378

24

Antihyperglycemic of Sargassum sp. Extract 381 Muhamad Firdaus, Rahmi Nurdiani, and Asep A. Prihanto

24.1 24.2 24.3 24.3.1

Introduction 381 Seaweed Bioactivities 382 In Vivo Hypoglycemic Activity of S. aquifolium Extract 384 Inhibition of α-Amylase and α-Glucosidase by Sargassum sp. Extracts 385 Area under Curve (AUC) 386 In Vivo Hypoglycemic Effects of S. aquifolium Extracts on Diabetic Rats 387 Body Weight 388 Blood Glucose 389 Hemoglobin A1 c (HbA1 c) 390 Conclusion 391 References 391

24.3.2 24.4 24.4.1 24.4.2 24.4.3 24.5

372

Contents

25

Immunological Activity of Marine Microalgae Extracts 395 Mariangela Caroprese, Maria G. Ciliberti, and Marzia Albenzio

25.1 25.1.1 25.1.2 25.1.3 25.2 25.2.1 25.2.2 25.2.3 25.3

Marine Microalgae Extracts 395 Phytosterols 398 Carotenoids and Vitamins 399 Polyunsaturated Fatty Acids 399 Overview of the Immune System 401 Immunological Activity of Sterols 402 Immunological Activity of Carotenoids and Vitamins Immunological Activity of Fatty Acids 406 Conclusion 407 References 407

26

Algal Polysaccharides and Their Biological Applications 411 Sirisha L. Vavilala and Jacinta S. D’Souza

26.1 26.2 26.2.1 26.2.1.1 26.2.1.2 26.2.1.3 26.2.2 26.2.2.1 26.2.2.2 26.2.3 26.2.3.1 26.3 26.3.1

Introduction 411 Algal Sulfated Polysaccharides 414 Sulfated Polysaccharides from Brown Algae 414 Alginates 414 Laminarin 415 Fucoidan 416 Sulfated Polysaccharides from Red Algae 418 Carrangeenans 418 Agar 419 Sulfated Polysaccharides from Green Algae 421 Ulvans 421 Applications of Bioactive Algal Polysaccharides 422 Anticoagulant and Antithrombotic Activities of Sulfated Polysaccharides 423 Antiviral Activities 427 Immunoin-flammatory Activities 429 Antioxidant Activities 431 Antilipidemic Activities 431 Sulfated Polysaccharides as Dietary Fibers 432 Seaweed Products and Potential of Its Biomass 433 Alginates 433 Agar 435 Mannitol 435 Seaweed Biomass for Bioenergy Production 435 Ethanol and Butanol from Brown Seaweeds 436 Future Prospects and Conclusion 438 References 438

26.3.2 26.3.3 26.3.4 26.3.5 26.3.6 26.3.7 26.3.7.1 26.3.7.2 26.3.7.3 26.3.8 26.3.8.1 26.4

405

IX

X

Contents

27

Biological Phlorotannins of Eisenia bicyclis Sang-Hoon Lee and Se-Kwon Kim

27.1 27.2 27.2.1 27.2.2 27.2.3 27.2.4 27.2.5 27.3

Introduction 453 Biological Activities of E. bicyclis 455 Antiviral Activity 455 Antioxidant Activity 456 Antitumor Activity 457 Anti-Inflammatory Activity 458 Antidiabetic Activity 460 Concluding Remarks 460 Acknowledgment 461 References 461

453

Part V: Biomedical Applications of Marine Algae 465 28

Algal Extracts as a Carrier of Micronutrients – Utilitarian Properties of New Formulations 467 Łukasz Tuhy, Katarzyna Chojnacka, Izabela Michalak, and Anna Witek-Krowiak

28.1 28.2 28.3 28.4

Introduction 467 The Application of Chelation Process in the Fertilizer Industry 467 Mechanism of Chelation 468 Seaweed Polysaccharides as a Source of Natural Chelators of Micronutrient Ions 468 Examples of Seaweed Polysaccharides – Potential Chelators of Microelement Ions 469 Alginate 469 Carrageenan 471 Ulvan 472 Fucoidan 473 Laminarin 474 Agar 475 Porphyran 475 Gel Formation by Seaweed Polysaccharides 476 Extraction Procedure of Polysaccharides 478 Examples of Chelating Properties of Extracted Seaweed Polysaccharides 479 New Approach toward Chelating Micronutrients by Polysaccharides 480 Regulations 482 Examples of Available Commercial Products 482 Conclusions 483 Acknowledgments 483 References 484

28.5 28.5.1 28.5.2 28.5.3 28.5.4 28.5.5 28.5.6 28.5.7 28.6 28.7 28.8 28.9 28.10 28.11 28.12

Contents

29

Marine Algae Based Biomaterials for Osteoblast Differentiation and Tissue Regeneration 489 Pathum Chandika and Won-Kyo Jung

29.1 29.2 29.3

Introduction 489 Scaffolds for Tissue Regeneration 490 Potentials of Marine Algae Derived Biomaterials for Bone Regeneration 492 Marine Algae Sauce for Bone Tissue Engineering 492 Algae Based Hydroxyapatite for Bone Tissue Engineering 498 Effects of Marine Algae on Osteoblast Differentiation 499 Osteoclast Inhibition through Marine Algae 500 Conclusions 501 Acknowledgments 502 References 502

29.3.1 29.3.2 29.4 29.5 29.6

30

Marine Algae Derived Polysaccharides for Bone Tissue Regeneration 509 Jayachandran Venkatesan and Se-Kwon Kim

30.1 30.2 30.2.1 30.2.2 30.3 30.3.1 30.3.2 30.3.3 30.4

Introduction 509 Alginate 511 Isolation Procedure of Alginate from Seaweed 511 Biomedical Application of Alginate 511 Fucoidan 513 Isolation of Fucoidan 514 Osteogenic Differentiation of Fucoidan 516 Fucoidan Composites for Bone Tissue Engineering 516 Conclusions 517 Acknowledgments 517 References 517

31

Wound Dressings from Algal Polymers 523 Monica Bhatnagar and Ashish Bhatnagar

31.1 31.2 31.3 31.4 31.5 31.5.1 31.5.1.1 31.5.1.2 31.5.1.3 31.5.1.4 31.5.1.5 31.5.1.6 31.5.2

Introduction 523 Wound 524 Wound Healing 525 Wound Dressings 527 Algal Polymers in Wound Management 527 Macroalgae 527 Alginates 528 Fucoidans 531 Carrageenan 535 Ulvans 537 Agar Agar 539 Laminarin 542 Microalgal and Cyanobacterial Polymers 543

XI

XII

Contents

31.6

Conclusion 544 References 545

32

Marine Algae and Chronic Diseases 557 Kalimuthu Senthilkumar and Se-Kwon Kim

32.1 32.2 32.3 32.4 32.4.1 32.4.2 32.4.3 32.4.4 32.4.5 32.4.6 32.4.7 32.5

Introduction 557 Marine Algae 558 Biological Activity of Marine Algae 559 Marine Algae on Chronic Diseases 560 Cardiovascular Disease 561 Diabetes 562 Arthritis 563 Osteoporosis 564 Neurodegenerative Diseases 564 HIV/AIDS 565 Anticancer 566 Conclusion 567 Acknowledgments 567 References 568

33

Algae Wastes Biomass: A New Class of Low-Cost Material with Potential Applications in Environmental Engineering 575 Laura Bulgariu and Dumitru Bulgariu

33.1 33.2 33.3

Introduction 575 Some Structural Characteristics of Algae Waste Biomass 577 Utilization of Algae Waste Biomass for Heavy Metals Removal in Batch Systems 580 Influence of Some Experimental Parameters on Biosorption Efficiency 580 Desorption and Reuse 588 Modeling of Biosorption Process of Heavy Metals on Algae Waste Biomass 589 Utilization of Algae Waste Biomass for Heavy Metals Removal in Continuous Systems 593 Conclusions 597 References 598

33.3.1 33.3.2 33.3.3 33.4 33.5

Part VI: Food and Industrial Applications of Marine Algae 603 34

Algae Extract as a Potential Feed Additive 605 Mariusz Korczyński, Zuzanna Witkowska, Sebastian Opaliński, ´ Marita Swiniarska, and Zbigniew Dobrzański

34.1 34.2 34.2.1

Introduction 605 Biologically Active Compounds Polysaccharides 606

606

Contents

34.2.2 34.2.3 34.2.4 34.2.5 34.2.6 34.2.7

Proteins 609 Polyunsaturated Fatty Acids (PUFAs) 610 Polyphenols 614 Pigments 615 Minerals 617 Other Biologically Active Compounds 617 Acknowledgments 617 References 617

35

Application of Marine Algae Derived Nutraceuticals in the Food Industry 627 Isuru Wijesekara and Se-Kwon Kim

35.1 35.2 35.2.1 35.2.2 35.2.3 35.2.4 35.2.5 35.2.6 35.2.7 35.3 35.3.1 35.3.2 35.3.3 35.3.4 35.4

Introduction 627 Bioactive Components from Marine Algae as Nutraceuticals 628 Phlorotannins 628 Sulfated Polysaccharides 628 Fucoxanthin and Astaxanthin 629 Lectins 631 Fucosterol 631 Mycosporine-Like Amino Acids 631 Proteins and Peptides 632 Health Beneficial Effects of Nutraceuticals from Marine Algae 632 Anticancer Effect 632 Antioxidant Effect 633 Anticoagulant Effect 633 Anti-HIV and Antimicrobial Effects 634 Concluding Remarks 634 References 635

36

Microalgal Carotenoids: Bioactive Roles, Health Foods, and Pharmaceuticals 639 J. Paniagua-Michel, Jorge Olmos Soto, and Eduardo Morales Guerrero

36.1 36.2 36.3 36.3.1 36.3.2 36.3.3 36.4 36.5 36.6 36.7

Introduction 639 Bioactive Roles of Microalgal Carotenoids 640 Microalgal Carotenoids as Food Additives 643 β-Carotene from Dunaliella salina 644 Astaxanthin from Haematococcus 644 Lutein from Chlorella 644 Carotenoids from Microalgae for Aquaculture 646 The Pro-vitamin A Bioactivity of Microalgae Carotenoids 647 Microalgal Carotenoids and Their Antioxidant Activity 648 Microalgae Carotenoids: Biomedical and Pharmaceutical Potential 650 Anticancer Properties of Microalgae Carotenoids 651 Carotenoids and Macular Degeneration 653

36.8 36.9

XIII

XIV

Contents

36.10

Conclusions 653 References 654

37

Biologically Active Organic Compounds, Especially Plant Promoters, in Algae Extracts and Their Potential Application in Plant Cultivation 659 Bogusława Górka, Jacek Lipok, and Piotr P. Wieczorek

37.1 37.2 37.2.1 37.2.2 37.2.3 37.2.4 37.2.5 37.3

Algae as a Source of Bioactive Substances 659 Plant Hormones and Hormone-Like Compounds in Algae 662 Auxins 663 Gibberellins 664 Cytokinins 665 Brassinosteroids 666 Other Compounds Regulating Plant Growth 666 Methods of Isolation and Fractionation of Active Compounds from Algal Extracts 668 Algal Extracts – Sample Preparation for Analytical Purposes 670 Quantitative and Qualitative Methods of Algal Active Compounds Determination 672 Application of Algae and Algal Originated Products in Agriculture 673 Perspectives 675 Acknowledgment 676 References 676

37.4 37.5 37.6 37.7

38

Biomass and Extracts of Algae as Material for Cosmetics 681 Joanna Fabrowska, Bogusława Łe˛ska, Grzegorz Schroeder, Beata Messyasz, and Marta Pikosz

38.1 38.2 38.2.1 38.2.2 38.2.3 38.2.4 38.2.5 38.3 38.3.1 38.3.2 38.3.3 38.4

Introduction 681 Bioactive Compounds 682 Polysaccharides 682 Proteins 686 Lipids 687 Pigments 688 Phenolic Compounds and Others 689 Application in Cosmetic Products 690 Algae Biomass 691 Algae Extracts 693 Quality Assurance and Regulations 698 Conclusion 701 Acknowledgments 701 References 701 Index 707

XVII

List of Contributors Helena T. Abreu

Monica Bhatnagar

ALGAplus–Produção e comercialização de algas e seus derivados, Lda., CIEMAR Travessa Alexandre da Conceição 3830-196 Ílhavo Portugal

Maharshi Dayanand Saraswati University Algae Biofuel and Biomolecules Centre Ajmer 305 009 India

Marzia Albenzio

Dumitru Bulgariu

University of Foggia Department of the Sciences of Agriculture Food and Environment (SAFE) Via Napoli, 25 71122 Foggia Italy

“Alexandru Ioan Cuza” University of Iasi Department of Geology Faculty of Geography and Geology Iasi Romania

Sekar Ashokkumar

Filial of Iasi Collective of Geography Romanian Academy Iasi Romania

The University of Suwon Department of Bioscience and Biotechnology Hwaseong-si 45-743 Republic of Korea

and

Laura Bulgariu Ashish Bhatnagar

Maharshi Dayanand Saraswati University Algae Biofuel and Biomolecules Centre Ajmer 305 009 India

Technical University Gheorghe Asachi of Iasi Department of Environmental Engineering and Management Faculty of Chemical Engineering and Environmental Protection D. Mangeron, No. 73 700050 Iasi Romania

XVIII

List of Contributors

Damien L. Callahan

Pathum Chandika

Deakin University Centre for Chemistry and Biotechnology School of Life and Environmental Science Burwood Victoria 3125 Australia

Pukyong National University Center for Marine-Integrated Biomedical Technology (BK21 Plus) Department of Biomedical Engineering Busan 608-737 Republic of Korea

and Metabolomics Australia The University of Melbourne The School of Botany Parkville Victoria 3010 Australia Mariangela Caroprese

University of Foggia Department of the Sciences of Agriculture Food and Environment (SAFE) Via Napoli, 25 71122 Foggia Italy Luciana R. de Carvalho

Instituto de Botânica Núcleo de Pesquisa em Ficologia Miguel Estéfano Ave, 3687 São Paulo, SP 04301–902 Brazil H.H Chaminda Lakmal

Jeju National University School of Marine Biomedical Sciences Department of Marine Life Science 1 Ara 1-dong, 102 Jejudaehakno Jeju 690-756 Republic of Korea

Katarzyna Chojnacka

Wroclaw University of Technology Institute of Inorganic Technology and Mineral Fertilizers Department of Chemistry ul. Smoluchowskiego 25 50-372 Wrocław Poland Maria G. Ciliberti

University of Foggia Department of the Sciences of Agriculture Food and Environment (SAFE) Via Napoli, 25 71122 Foggia Italy Rosalia Contreras

Centro de Investigación Cient´ıfica y de Educación Superior de Ensenada (CICESE) Molecular Microbiology Laboratory Department of Marine Biotechnology Carretera Ensenada-Tijuana No. 3918 Zona Playitas Ensenada, BC C.P. 22860 Mexico


Agnieszka De˛bczak

Zbigniew Dobrzañski

New Chemical Syntheses Institute Supercritical Extraction Department Aleja Tysia˛clecia Państwa Polskiego 13a 24-110 Puławy Poland

Wroclaw University of Environmental and Life Science Department of Environment Animal Hygiene and Animal Welfare Chełmoñskiego 38C 51-631 Wrocław Poland

Pradeep Dewapriya

Agnieszka Dobrzyñska-Inger

Pukyong National University Marine Biochemistry Laboratory Department of Chemistry Busan 608-737 Republic of Korea


Daniel A. Dias

Metabolomics Australia The University of Melbourne The School of Botany Parkville Victoria 3010 Australia and Deakin University, Centre for Chemistry and Biotechnology School of Life and Environmental Science Burwood Victoria 3125 Australia Agnieszka Dmytryk

Wrocław University of Technology Institute of Inorganic Technology and Mineral Fertilizers Department of Chemistry ul. Smoluchowskiego 25 50-372 Wrocław Poland

Jacinta S. D’Souza

UM-DAE Centre for Excellence in Basic Sciences Department of Biology Vidyanagari, UM Campus Kalina, Santacruz (E) Mumbai 400098 India Joanna Fabrowska

Adam Mickiewicz University in Poznań Department of Supramolecular Chemistry Faculty of Chemistry Umultowska 89b 61-614 Poznań Poland

XIX

XX


Julyana N. Farias

Mateusz Gramza

Post-Graduate Program in “Biodiversidade Vegetal e Meio Ambiente” Instituto de Botânica Miguel Estéfano Ave, 3687 São Paulo, SP 04301–902 Brazil

Biotek Agriculture Gać 64 55-200 Oława Poland

Muhamad Firdaus

Brawijaya University Department of Biochemistry Faculty of Fisheries and Marine Sciences Malang East Java 65145 Indonesia Mutue T. Fujii

Institute of Botany Research Center in Phycology Miguel Estéfano Ave, 3687 São Paulo, SP 04301–902 Brazil

Eduardo Morales Guerrero

Centro de Investigación Cient´ıfica y de Educación Superior de Ensenada (CICESE) Department of Marine Biotechnology Ensenada BC 22860 Mexico David R.A. Hill

The University of Melbourne The Department of Chemical and Biomolecular Engineering Parkville Victoria 3010 Australia You-Jin Jeon

Henryk Górecki


Jeju National University School of Marine Biomedical Sciences Department of Marine Life Science 1 Ara 1-dong, 102 Jejudaehakno Jeju 690-756 Republic of Korea Won-Kyo Jung

Bogusława Górka

Opole University Department of Analytical and Ecological Chemistry Faculty of Chemistry Pl. Kopernika 11 a 45-040 Opole Poland

Pukyong National University Center for Marine-Integrated Biomedical Technology (BK21 Plus) Department of Biomedical Engineering Busan 608-737 Republic of Korea


and

and

Chosun University Department of Marine Life Science 375 Seosuk-Dong Dong-Gu Gwangju 501-759 Republic of Korea

Pukyong National University Marine Biochemistry Laboratory Department of Chemistry 599-1 Daeyeon 3-dong, Nam-gu Busan 608-737 Republic of Korea

Fatih Karadeniz

Mariusz Korczyñski

Pukyong National University Marine Bioprocess Research Center 599-1 Daeyeon 3-dong Nam-gu Busan 608-737 Republic of Korea


Mustafa Z. Karagozlu

Pukyong National University Marine Bioprocess Research Center 599-1 Daeyeon 3-dong Nam-gu Busan 608-737 Republic of Korea Keun Kim

The University of Suwon Department of Bioscience and Biotechnology Hwaseong-si 45-743 Republic of Korea Se-Kwon Kim

Pukyong National University Marine Bioprocess Research Center Specialized Graduate School Science and Technology Convergence Marine Biotechnology Laboratory Department of Chemistry Department of Marine-Bio Convergence Science 599-1 Daeyeon 3-dong, Nam-gu Busan 608-737 Republic of Korea

Dorota Kostrzewa

New Chemical Syntheses Institute Supercritical Extraction Department Aleja Tysia˛clecia Państwa Polskiego 13a 24-110 Puławy Poland Sang-Hoon Lee

Korea Food Research Institute Baekhyun-dong Seongnam Gyeonggi 463-746 Republic of Korea and University of Science and Technology Pukyong National University Marine Bioprocess Research Center, Specialized Daejeon 305-350 Republic of Korea

XXI

XXII


Bogusława Łe˛ska

Panchanathan Manivasagan


Pukyong National University Marine Bioprocess Research Center Department of Chemistry Marine Biotechnology Laboratory 599-1 Daeyeon 3-dong Nam-gu Busan 608-737 Republic of Korea

Yong Li

Changchun University of Chinese Medicine Department of Pharmaceutical Sciences, 1035, Boshuo Road Jing Yue Economic Development Zone Chanchun City Jilin Province People’s Republic of China

Gregory J.O. Martin

The University of Melbourne The Department of Chemical and Biomolecular Engineering Parkville Victoria 3010 Australia Filipa Meireles

Yong-Xin Li

Marine Bioprocess Research Center Pukyong National University Busan 608-737 Republic of Korea Jacek Lipok

Opole University Department of Analytical and Ecological Chemistry Faculty of Chemistry Pl. Kopernika 11 a 45-040 Opole Poland Kuppusamy Manimaran

Annamalai University Marine Biology, Centre of Advanced Study in Marine Biology Faculty of Marine Science Parangipettai 608 502 Tamil Nadu India

University of Coimbra IMAR-CMA Department of Life Sciences Faculty of Sciences and Technology Rua da Matemática, n∘ 49 3001-455 Coimbra Portugal Beata Messyasz

Adam Mickiewicz University in Poznań Department of Hydrobiology Faculty of Biology Umultowska 89 61-614 Poznań Poland


Izabela Michalak

Ian L.D. Olmstead


The University of Melbourne The Department of Chemical and Biomolecular Engineering Parkville Victoria 3010 Australia Sebastian Opaliñski

Marcin Mikulewicz

Wroclaw Medical University Department of Dentofacial Orthopedics and Orthodontics ul. Krakowska 26 50-425 Wrocław Poland


Rahmi Nurdiani

Brawijaya University Department of Fishery Product Technology Faculty of Fisheries and Marine Sciences Malang East Java 65145 Indonesia Tatsuya Oda

Nagasaki University Division of Biochemistry Faculty of Fisheries Bunkyo-machi 1-14 Nagasaki 852-8521 Japan Jorge Olmos Soto

Centro de Investigación Cient´ıfica y de Educación Superior de Ensenada (CICESE) Molecular Microbiology Laboratory Department of Marine Biotechnology Carretera Ensenada-Tijuana No. 3918 Ensenada, Baja California C.P. 22860 Mexico

J. Paniagua-Michel

Centro de Investigación Cient´ıfica y de Educación Superior de Ensenada (CICESE) Bioactive Compounds and Bioremediation Laboratory Department of Marine Biotechnology Carretera Ensenada-Tijuana No. 3918 Zona Playitas Ensenada C.P. 22860 Mexico Ratih Pangestuti

Research Center for Oceanography Indonesian Institute of Sciences Jl. Pasir Putih 1, Ancol Timur Jakarta Utara 14430 Republic of Indonesia

XXIII

XXIV


Leonel Pereira

Edward Rój

IMAR-CMA/MARE Department of Life Sciences Faculty of Sciences and Technology University of Coimbra Calçada Martim de Freitas 3000-456 Coimbra Portugal


Marta Pikosz

Viviana P. Rubio

Adam Mickiewicz University in Poznań Department of Hydrobiology Faculty of Biology Umultowska 89 61-614 Poznań Poland

Universidad Autónoma de Baja California (UABC) Ensenada Marine Science Faculty Km 103 Carretera Tijuana-Ensenada Ensenada, Baja California C.P. 22860 México

Asep A. Prihanto

Brawijaya University Department of Fishery Product Technology Faculty of Fisheries and Marine Sciences Malang East Java 65145 Indonesia

Agnieszka Saeid

Wroclaw University of Technology Institute of Inorganic Technology and Mineral Fertilizers Faculty of Chemistry ul. Smoluchowskiego 25 50-372 Wrocław Poland

Paulo J.A. Ribeiro-Claro

University of Aveiro CICECO Department of Chemistry 3810-193 Aveiro Portugal Pablo Riul

Universidade Federal da Para´ıba Departamento de Engenharia e Meio Ambiente, CCAE 58297-000 Rio Tinto, PB Brazil

Kalpa W. Samarakoon

Jeju National University School of Marine Biomedical Sciences Department of Marine Life Science 1 Ara 1-dong, 102 Jejudaehakno Jeju 690-756 Republic of Korea


Grzegorz Schroeder

Marita S´winiarska



Kalimuthu Senthilkumar

Łukasz Tuhy

Pukyong National University Marine Bioprocess Research Center Department of Chemistry Marine Biotechnology Laboratory Busan 608-737 Republic of Korea


Adam Słowiński

Mikinori Ueno

Arysta LifeScience Poland Przasnyska 6B 01-756 Warszawa Poland

Nagasaki University Division of Biochemistry Faculty of Fisheries Bunkyo-machi 1-14 Nagasaki 852-8521 Japan

Katarzyna Ste˛pnik

Maria Curie-Skłodowska University Department of Planar Chromatography Faculty of Chemistry Maria Curie-Skłodowska Square 3 20-031 Lublin Poland

Sirisha L. Vavilala

UM-DAE Centre for Excellence in Basic Sciences Department of Biology Vidyanagari, UM Campus, Kalina Santacruz (E) Mumbai 400098 India

XXV

XXVI


Jayachandran Venkatesan

Radoslaw Wilk

Pukyong National University Marine Bioprocess Research Center Department of Marine Bio Convergence Science 599-1 Daeyeon 3-dong Busan 608-737 Republic of Korea

Wroclaw University of Technology Institute of Inorganic Technology and Mineral Fertilizers Department of Chemistry ul. Smoluchowskiego street 25 50-372 Wrocław Poland

Piotr P. Wieczorek

Anna Witek-Krowiak

Opole University Department of Analytical and Ecological Chemistry Faculty of Chemistry Pl. Kopernika 11 a 45-040 Opole Poland

Wrocław University of Technology Department of Chemistry Division of Chemical Engineering Norwida 4/6 50-373 Wrocław Poland

Isuru Wijesekara

KU Leuven Toxicology and Pharmacology Herestraat 49 Leuven 3000 Belgium and Pukyong National University Department of Chemistry Busan 608-737 Republic of Korea

Zuzanna Witkowska

Wroclaw University of Technology Institute of Inorganic Technology and Mineral Fertilizers Smoluchowskiego 25 50-373 Wrocław Poland

XXVII

Preface Marine algae are popular food ingredients mainly in Asian countries such as Korea, Japan, and China. They are also well known for their health benefits because of the presence of bioactive components in them. Marine algae are rich in vitamins, minerals, dietary fibers, proteins, polysaccharides, and various functional polyphenols. Recently, several studies have demonstrated the variety of biological benefits associated with marine algal polyphenols, including antioxidant, anticoagulant, antibacterial, anti-inflammatory, and anticancer activities. These marine macroalgae have been classified based on pigmentation into brown (Phaeophyta), red (Rhodophyta), and green (Chlorophyta) types. Apart from food uses, including their main industrial use as thickeners and gelling agents, seaweeds are used widely as ingredients in nutraceutics and cosmetics and as fertilizers. Marine Algae Extracts – Processes, Products, and Applications describes the characteristic features of marine algae cultivation, identification, production, process, and applications (biological, biomedical, food, and industrial). The book is divided into six parts: Part I provides the cultivation and identification processes of marine algae; Part II discusses the production and processing of marine algae; Part III provides an overview of the marine algae products; Part IV discusses the various biological applications of marine algae; Part V analyzes the numerous biomedical applications of marine algae; and Part VI examines the food and industrial applications of marine algae. Each part is a collection of comprehensive information on the past and present research on marine algae, compiled by proficient scientists worldwide. I personally intend to mention that the findings and the recent information provided in this book will be helpful to the upcoming researchers to establish a phenomenal investigation from a wide range of research areas. I hope that the fundamental as well as applied contributions in this book serve as a potential research and development leads for the benefit of humankind. Altogether, marine algal biotechnology will be the hottest field in future toward

XXVIII

Preface

the enrichment of targeted algal species, which further establishes a sustainable oceanic environment. This book would be a reference book for the emerging students in the academic and industrial research. Busan, South Korea 10 Nov 2014

Se-Kwon Kim

XXIX

Acknowledgments I would like to thank Wiley-Blackwell Publishers for their encouragement and suggestions to get this wonderful compilation published. I would also like to extend my sincere gratitude to all the contributors for providing help, support, and advice to accomplish this task. Further, I would like to thank Dr. Panchanathan Manivasagan and Dr. Jayachandran Venkatesan, who worked with me throughout the course of this book project. I strongly recommend this book for marine algae extracts researchers/students and hope that it helps to enhance their understanding in this field. Se-Kwon Kim & Katarzyna Chojnacka

1

1 Introduction of Marine Algae Extracts Katarzyna Chojnacka and Se-Kwon Kim

1.1 Introduction

Recently, there is increased interest in naturally produced active compounds as alternatives to synthetic substances. Although these compounds often show lower activity, they are nontoxic and do not leave residues. It has already been reflected by the projects of new law regulations in EU countries that have imposed legal restrictions on the use of xenobiotics as plant protection products or preservatives. In the European Union there are plans of new directives that impose additional environmental taxes, primarily because of the residues of active substances in the environment. This implies that there is a need to develop new and safe products of biological origin, with properties similar to the synthetic, in particular antimicrobial, antifungal, antioxidizing compounds, and colorants. These natural compounds are found in algal extracts (Table 1.1). Algal biomass have been used for centuries as food and medicine. The health promoting effects of algae were discovered as early as 1500 BC [1]. However, the biomass of algae gained interest as a source of chemicals and pharmaceuticals only recently. Nowadays, the production regime requires the use of extracts rather than the biomass itself, because of the formulation requirements (consistency, stability, color, flavor, etc.). Until now, algal products were available mainly as tablets, capsules, or liquid extracts, and sometimes were incorporated into food products, cosmetics, or products for plants [2]. In 2006, the market of microalgal biomass produced 5000 mg dry biomass/year and generated a turnover of 1.25 × 109 USD [2]. The global sector of macroalgae is worth 6 billion USD, with main contribution from hydrocolloids and crop protection products [3]. Recently, compounds derived from algae (carotenoids, β-carotene astaxanthin, long-chain polyunsaturated fatty acids (PUFAs), docosahexaenoic acid) began to be produced on industrial scale [4]. Novel compounds isolated from algae possess a great further potential to be applied for their pharmacological and biological activity [4]. Seaweeds produce a vast spectrum of secondary metabolites because they live in nonfriendly environment but are not damaged photodynamically as they synthesize protective compounds and develop protecting mechanisms Marine Algae Extracts: Processes, Products, and Applications, First Edition. Edited by Se-Kwon Kim and Katarzyna Chojnacka. © 2015 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2015 by Wiley-VCH Verlag GmbH & Co. KGaA.

2

1 Introduction of Marine Algae Extracts

Table 1.1 Major compounds in algal extracts [2, 11, 19, 20]. Compound

Function

Application

Polysaccharides

Components of cell wall (fucoidan, alginate, laminarin) Not found in terrestrial plants Phenol rings in polyphenols act as electron traps to scavenge radicals

Provide strength, flexibility, maintain ionic equilibrium, prevent from desiccation Antimicrobial, antioxidant, antiviral compounds that protect the algae from abiotic and biotic stress conditions, for example, phlorotannins that are formed from oligomeric structures and phloroglucinol Antioxidative, but difficult to extract Structural membrane lipids; important in human and livestock diet. Composed of glycerol, sugars, bases esterified with fatty acids (saturated or unsaturated (C12–C22)) Antioxidant, antiviral, anti-inflammatory activity, UV protection —

Phenolics and phlorotannins

Protein, peptides, and essential amino acids Lipids

The contents vary

Terpenoids and steroids

Carotenoids, xanthophyll, fucoxanthin, astaxanthin

Vitamins

A, B1, B2, B6, B12, C, E, nicotinate, biotin, folic acid, pantothenic acid. Level depends on the season Se, Zn, Mn, Cu – structural components of antioxidative enzymes

Minerals

Polyunsaturated fatty acids (PUFA) (ω-3 and ω-6) – higher level than in terrestrial plants

—

[5]. Environmental stress to which algae are exposed include rapid fluctuations of light intensity, temperature, osmotic stress, desiccation that lead to the formation of free radicals and oxidizing agents that lead to photodynamic damage [6].

1.2 Algal Biomass as a Useful Resource

Algae are the oldest photosynthetic organisms dating back to 3.8 billion years (prokaryotic cyanophytes) [7]. The number of species is estimated as 280 000 [7]. Algal biomass is being used as the raw material for different branches of industry

1.2

Algal Biomass as a Useful Resource

and the global production is prevalently increasing [7]. Algae are photosynthetic organisms that convert light energy from the Sun into chemical energy stored in the form of chemical compounds in the process of photosynthesis [1]. A characteristic of algae is that they possess a simple reproductive structure [8]. The biomass of algae contains various compounds with diversified structures and functions that are synthesized in the response to stress conditions, for example, heat/cold, desiccation, salinity, osmotic stress, anaerobiosis, nitrogen deficiency, photooxidation, as protection from physiological stressors [1]. Algae are a diversified group of organisms and are divided into microalgae and macroalgae. The first group includes prokaryotic cyanobacteria and eukaryotic microalgae [9]. Algae are very diversified organisms when considering size (from unicellular microalgae to multicellular macroalgae) [10]. The basis for the classification of algae is pigmentation: green (Chlorophyceae), red (Rhodophyceae), and brown (Phaeophyceae) [11]. The difference concerns not only pigmentation, but also the type of storage material and the composition of cell wall polysaccharides [12]. Algae are simpler than terrestrial plants [12]. Algae could be considered as natural factories that produce bioactive compounds [13]. The composition of green algae: 10% protein, 35% carbohydrate, and 50% ash (Ca, Fe, P, Cl) [12]. Algae were in use since prehistory as the components of diet and as medicine [14]. Although the importance of algal industry is permanently increasing, there are some contradictions between Asian (Far East) and European ways of utilization of this resource [14]. In Europe, the biomass of seaweeds was treated as a sort of waste from seas and oceans [14]. Certainly, algal biomass is still an underutilized biological resource. Algal biotechnology is divided into two branches: microalgal and macroalgal, with its unique specificity [15]. Microscopic algae are called microalgae; however, this term is not related to taxonomy. Among microalgae, cyanobacteria are distinguished, which are prokaryotic [15]. Macroalgal biotechnology includes the production of (phycocolloids agar-agar, alginates, carrageenan) from Rhodophyta and Phaeophyta, and the global value is 6 × 109 per year [15]. At present, the main directions in macroalgal biotechnology are biofuels, agricultural biostimulants for crop plants, probiotics for aquaculture, soil bioremediation, wastewater treatment, and biomedical applications of extracted compounds (polyphenols, polysaccharides) [3]. Microalgal biotechnology refers to the production of different products: phycocyanin, carotenoids (β-carotene, astaxanthin), fatty acids and lipids, polysaccharides, immune modulators that find an application in health food, cosmetics, feed and food supplements, pharmaceuticals, and fuel production [15]. Microalgal groups of the major importance are cyanobacteria (Spirulina sp.), Chlorophyta (Chlorella sp., Dunaliella sp.), Rhodophyta (Porphyridium sp.), Bacillariophyta (Odontella sp., Phaeodactylum sp.) [15]. While macroalgae are harvested from natural habitats, microalgae are cultivated in artificial systems [15]. The products of microalgal biotechnology are coloring substances (astaxanthin, phycocyanin, phycoerythrin), antioxidants (β-carotene, tocopherol, antioxidant CO2 extract), and arachidonic acid (ARA), docosahexaenoic acid (DHA), and PUFA extracts [15].

3

4


1.3 Biologically Active Compounds Extracted from Algae

Because algae are coastal primary producers and have impressive possibilities to survive in extreme environmental conditions, in particular to trigger oxidative stress, they produce a variety of useful compounds [16]. Algae live in extreme conditions: fluctuating salinity, temperature, nutrients, and UV–vis irradiation [10]. Long periods of desiccation cause overproduction of reactive oxygen species, which is neutralized by physiological and biological mechanisms: the production of secondary metabolites [16]. Therefore, compounds isolated from the biomass of seaweeds possess biological activity. The biomass of algae contains many valuable components: minerals, vitamins (A, B, C, E), PUFAs (ω-3), amino acids, proteins, polysaccharides, lipids, and dietary fiber [17]. Many of these bioactive constituents can be extracted to obtain antioxidative, anti-inflammatory, antimicrobial, anticancer, antihypertensive products [11, 17]. Particularly useful are secondary metabolites with antiviral, antimalarial, anticancer properties [1]. Products derived from algae also contain polysaccharides, polyphenolic compounds, and terpenes [11]. Seaweeds and their extracts are added to food as antioxidants, antimicrobials, dietary fiber, and dietary iodine [6]. In various studies, strong antioxidative properties of compounds isolated from seaweeds were confirmed [18]. Antioxidative activity produces phlorotannins (polyphenolic compounds – 1–10% d.m. of brown seaweeds), alkaloids, terpenes, ascorbic acid, tocopherols, and carotenoids [18]. Antioxidants transform radicals into nonradicals by donating electrons and hydrogen, chelation of transition metals, and dissolving peroxidation compounds [6]. The role of antioxidants is to prevent lipid oxidation, inhibiting the formation of products as a result of oxidation, and consequently prolonging the shelf life of products [6]. Algae are a rich source of natural antioxidants and antimicrobial compounds [6]. The research on the composition of algal extracts concerns mainly antioxidants as an alternative to synthetic, because according to recent research these compounds if used as food additives are potential promoters of carcinogenesis [1]. The extracts modulate the oxidative stress and diseases related to radical scavenging effect: sesquiterpenoids and flavonoids (green alga Ulva lactuca), phlorotannins (brown alga Eisenia bicyclis, Ecklonia cava, E. kurome), phycobiliprotein, and phycocyanin (blue-green alga Spirulina platensis), which protect from DNA damage by H2 O2 [17].

• Anti-HIV – cyanovirin – protein from Nostoc ellipsosporum [1] • Photoprotective compounds – repair DNA damage – mycosporine-like amino acids, scytonemin enzymes (shock proteins) – superoxide dismutase, catalase, and peroxidase [1]. Microalgae contain carotenoids, PUFAs, phycobilins, sterols, polyhydroxyalkonates, and polysaccharides [9]. They can be considered as cosmeceuticals, nutraceuticals, and functional foods [9]. For instance, Spirulina contains lipids

1.4

The Application of Products Derived from Algal Biomass

(6–13; 50% in the form of fatty acids), phycocyanin (20–28%), and carbohydrates (15–20%; mainly as polysaccharides) [21]. Algal cells contain phytochelatins – proteins synthesized in response to exposure to toxic metal ions [22]. However, the attempt to extract and use these proteins is not found in the available literature [22].

1.4 The Application of Products Derived from Algal Biomass

The global wild stocks of seaweeds yield 8 million mg of biomass [18]. In 2004, the contribution in the market was as follows: sea vegetables (88%), phycocolloids (11%), phycosupplements (1%), and the minor contribution of soil additives, agrochemicals, and animal feeds (totally, 6000 million USD) [14]. Algal extracts create a new market sector, because they can be used in a variety of products, for example, antioxidant capsules containing Spirulina extract, Chlorella extract in health drinks, oral capsules containing carotenoid extracts from Dunaliella [15]. Other examples of algal extracts-based products are pet functional food, biofertilizers (which increase water-binding capacity and serve as the source of minerals and substances promoting germination, growth of leaves and stems and flowering). Of particular interest are antioxidants present in algae and their extracts, as the use of synthetic antioxidants has been restricted because of toxicity and health risks [23]. It is important to replace these synthetic compounds with natural antioxidants [23]. Antioxidative compounds from marine sources include various functional compounds, for example, tocopherols [19]. Lipid-soluble algal extracts can be used as protective functional ingredients [19]. Antioxidative properties of natural compounds from algae can prolong the shelf life of foods and cosmetics through delayed oxidation [11]. Natural anti-oxidants may also be useful in treating aging, UV-exposure, and diseases associated with oxidation [11]. Extracts from algae are used in cosmetics, for example, from Spirulina and Chlorella [2]. Polysaccharides isolated from algae are other important components of foods and cosmetics and in nutraceutical and pharmaceutical preparations and are produced mainly from seaweeds [21]. Polysaccharides (carrageenans, alginates) are used in food industry as edible packaging materials [6]. The main source of industrially exploited polysaccharides (alginate, agar, carrageenan) originates from the biomass of algae [12]. Algal biomass contains significantly higher levels of polysaccharides than terrestrial plants [12]. Algal polysaccharides differ from those in terrestrial plants: sulfate groups, additional sugar residues, high content of ionic groups, high solubility in water, and unique rheological properties [12]. Polysaccharide production includes the following steps: selection of raw material, stabilization and grinding of biomass, extraction and purification, precipitation, and drying [12].

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1.4.1 Agriculture – For Plants

In modern agriculture, higher production should accompany lower environmental impact and higher sustainability [24]. These criteria fulfill biostimulants that improve efficiency of regular fertilization (increase the efficiency of nutrients uptake), enhance yield and the quality of crops, improve tolerance to environmental stress, and possess antioxidant properties [24]. Biostimulants are natural substances that promote growth, uptake of nutrients, and tolerance to abiotic stress and different climatic conditions [25]. Seaweed extracts can be used as foliar sprays for vegetables, grains, and flowers [24]. Plant growth regulators are defined as bioactive compounds. It is desired that they perform well and are degraded into products that are not harmful to the environment [26]. European Biostimulant Industry Council (EBIC) was established to help introduce agriculture biostimulants to the market and support regulatory EU authorities to describe biostimulants as innovative class of products, the production of which uses minimal synthetic processing. Biostimulants are approved in organic crops, with an important group of products derived from macroalgae [27]. Seaweeds have been used in the cultivation of plants since antiquity [28]. Seaweeds were composted since antiquity and used as soil amendments. The first industrial applications of seaweeds in agriculture were in 1944, as the new source of fiber [14]. At present, the extracts are applied directly to shoots foliarly or to soil [3]. The examples of algal extracts currently available on the market are Kelpak, Actiwave, and AlgaGreen [3]. Seaweed concentrates (e.g., Kelpak) are applied at low rates and have growth promoting effect following the presence of plant growth regulators (e.g., cytokinins and auxins, polyamines, putrescine, spermine) rather than nutrients [29]. These active substances increase the growth of nutrient-stressed plants [29]. In 1949, the product Maxicrop was developed [14]. Using liquid seaweed is advantageous, because plants respond immediately and positively (dilution 1 : 500); also, the ions of micronutrients (Cu, Co, Mn, Fe) are soluble at high pH and are chelated by partly hydrolyzed sulfated polysaccharides; soil crumb structure is improved (with alginate and fucoidan), microorganisms, root system, and plant growth are stimulated [14]. Extracts from seaweeds are useful in the cultivation of plants because they improve a wide range of physiological responses: increase crop yield, improve growth, improve plants’ resistance to frost, serve as biofungicide and bioinsecticide, increase nutrients’ uptake from soil because they contain plant growth regulators [30]. The extracts are used in low doses (high dilutions), because the active substances are efficient even in small quantities [30]. The compounds found in algal extracts that are important for plant growth are cytokinins, auxins, abscisic acid, vitamins, amino acids, and nutrients [24]. The outcome is the result of the synergistic effect of many compounds present in algal extracts [24]: phytohormones, betaines (organic osmolytes), polymers, nutrients, and alginic acid (soil conditioning agent that supports soil structure) [25, 28].

1.4

The Application of Products Derived from Algal Biomass

There are various reports of laboratory, pot, and field studies that aimed to test the plant growth stimulating properties of algal extracts. El-Baky et al. [31] investigated the effect of treatments with microalgae extracts (Spirulina maxima and Chlorella ellipsoida) on antioxidative properties in grains of wheat. The content of carotenoid, tocopherol, phenolic, and protein in grain was investigated. Antioxidant activity of ethanolic extracts showed the significant increase of radical scavenging activity in response to microalgal extracts treatment [31]. 1.4.2 Functional Food

Functional food is defined as food that positively affects one or more physiological functions to increase the well-being and reduce the risk of suffering for diseases [8]. Recently, a new market for functional food has evolved, the food called “food for the not-so-healthy” [13]. Functional food is produced by the addition of active components. Functional food contains functional ingredients: micronutrients ω-3 fatty acids, linoleic acids, phytosterols, soluble fiber (inulin – prebiotics), probiotics, carotenoids, polyphenols, vitamins that present healthy effect on the organism [13]. New, biologically active natural ingredients (antioxidant, antiviral, antihypertensive) extracted from the biomass of algae are becoming important research objects in the area of food science and technology [10]. Algal extracts are the components of functional food, because they are considered as natural, biologically active components. The latter, beside nutrition, should have the beneficial influence on functions of the body by improving health or preventing from diseases [32]. Extracts from Spirulina can be added to functional foods because of antioxidant, antimicrobial, anti-inflammatory, antiviral, and antitumoral properties of the compounds (phycocyanins, carotenoids, phenolic acids, and ω-3 and six PUFAs) [32]. Algae are used as dietary supplements that are classified into three groups: (i) Spirulina platensis, (ii) Aph. flos-aquae, and (iii) Chlorella pyrenoidosa [33]. The biomass of these microalgae is obtained either from lakes or by cultivation in artificial ponds [33]. Algae can be cultivated, in which the growth rate is high and in some cases there is a possibility of controlling the production of active compounds by adjusting cultivation conditions [10]. The potential use of brown seaweed extracts to inhibit the growth of microorganisms responsible for food spoilage and pathogenic microorganisms was also investigated [5]. The addition of 6% of the extract substantially reduced the growth of nondesired microflora [5]. 1.4.3 Cosmetics

Microalgae, the biomass of which is to be used as the raw material for isolation of beneficial compounds, are cultivated in artificial systems that provide the biomass that is free of impurities [7]. Algal extracts are useful in the skin care market as

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well because they support regeneration of tissues and reduce wrinkles, in particular, the extracts from Spirulina (which repair signs of aging, prevent stria formation) and Chlorella (stimulate collagen synthesis) [2]. The properties of microalgal extracts include reduction of intracellular oxidative stress and synthesis of collagen [7]. Extracts from the following microalgae are produced commercially for cosmetic industry [7]:

• Nannochloropsis oculata – vitamin B12, vitamin C, and antioxidants • Dunaliella salina – pigment industry (carotenes), amino acids, and polyphenols

• Chlorella vulgaris – proteins, and inorganics substances. 1.4.4 Pharmaceuticals

Algal extracts can replace commercial antibiotics in disease treatments [34]. Biologically active metabolites isolated from marine algae have the potential to be used as pharmaceuticals because they inhibit the growth of bacteria, viruses, and fungi [34]. The chemicals are macrolides, cyclic peptides, proteins, polyketides, sesquiterpenes, terpenes, and fatty acids [34]. Cavallo et al. [34] investigated the effect of lipid extracts from six algae and their antibacterial activity against fish pathogens and found that they can be used as antibacterial, health promoting feed for aquaculture. Extracts from Spirulina are active against viruses (herpes, influenza, cytomegalovirus) and inhibit carcinogenesis [35]. Spirulina is the source of vitamin A that is highly absorbable [36]. Hot water extract from Spirulina supports human immune system by the improvement of immune markers in blood (higher level of gamma interferon and interleukin-12p40 and toll-like receptors) and acts directly on myeloid lineages and natural killer-cells (NK cells) [35]. Immulina is a polysaccharide found in the extract from Spirulina that activates monocytes. Water extracts also showed antiviral activity [35]. 1.4.5 Fuels

Seaweed extracts can be the resource to produce liquid fuels (ethanol), because of high carbohydrates (laminaran, mannitol) content [37]. Seaweeds can be bioconverted to methane [37]. 1.4.6 Antifouling Compounds

Extracts from marine algae (e.g., Enteromorpha prolifera) contain compounds that have antifouling properties toward, for example, mussels (Mytilus edulis) and

1.5

Extraction Technology

larval settlement: tannins (Sargassum natans), bromophenol (Rhodomela larix), diterpenes (Dictyota menstrualis), and halogenated furanones (Delisea pulchra). These compounds have the potential in the prevention from fouling of ship hulls and aquaculture nets instead of organotin or paints based on toxic metals [38].

1.5 Extraction Technology

Seaweed industry was established in 1950s [3]. The production concerned mainly low-cost fertilizers and food [3]. For the first time liquefaction of seaweeds was undertaken in 1857 by compressing [28]. The goal was to obtain the formulation that is transportable over long distances [28]. Algal extracts were obtained and patented in 1952 by alkaline extraction [3]. Another process was milling in low temperature [28]. Although natural extracts possess a great applicable potential, the problem with natural products is variable composition of extracts because of fluctuations in the raw material (season, location), different extraction techniques [12]. Extraction methods vary and the following can be distinguished: ethanol, methanol, enzymatic [17], composting, supercritical CO2 extraction with cosolvents. In the elaboration of a new extraction technology, it is necessary to select the target bioactive compound, select the species of alga for extraction containing the compound of interest, select the operation conditions to find a compromise between the yield and purity, and consider if large enough resources of the algae are available. It is essential to develop appropriate, quick, cost-efficient, and environmentally friendly methods of extraction that aim to isolate biologically active compounds of interest [10] without loss of their activity. It is essential to develop extraction procedures that involve the use of specific solvents and processes [8]. The production of algal extracts consists of several unit operations [7]:

• Upstream processing – preparation for cultivation • Cultivation – in photobioreactors • Downstream processing – cell harvesting, rehydration and hot water extraction, centrifugation, and ultrafiltration

• Formulation, preservation, and conditioning. Traditional extraction techniques (soxhlet) solid–liquid extraction (SLE), liquid–liquid extraction (LLE) consume large quantities of solvents and require high extraction times [8]. These procedures present low yield of extraction and low selectivity toward bioactive compounds [8]. Because of the lack of automation, reproducibility is low [8]. Recently developed techniques supercritical fluid extraction (SFE), pressurized liquid extraction (PLE), accelerated solvent extraction (ASE), pressurized hot water extraction (PHWE), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) have further reduced these limitations [8]:

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10


• Solvent extraction – large quantities of toxic organic solvents are used, long time of extraction, laborious, low selectivity, low extraction yields, and not mild conditions (temperature, light, oxygen) [32]. • Pressure liquid extraction – less solvent, shorter time of extraction, automated, no oxygen, and no light [32]. • Supercritical fluid extraction – technique used to isolate active components from natural materials [32]. SFE uses solvents at temperatures and pressures above their critical point and is used to extract compounds from biomasses [8]. In this technique, the consumption of toxic organic solvents is reduced and the main solvent used is CO2 [8]. The disadvantage is low polarity of CO2 and resulting necessity of the use of polar modifiers or cosolvents [8]. Advantages are high diffusivity, easiness in the control of temperature and pressure (possibility of modification of solvent strength), and obtaining solvent-free extracts [8]. Extraction of biologically active compounds from algal biomass is not selective. The extract is a mixture of different compounds [11]. The factors that influence the composition and thus the activity of algal extracts depend on species, environmental conditions, season of the year, age, geographical location, and processing technologies [11]. For instance, ethanol was found to be more efficient in the extraction of polyphenols than water [23]. Seaweed extracts contain PUFAs (in particular ω-3 long chain PUFA) that have several health promoting effects and have the potential to be useful in treatment or reducing symptoms of: cardiovascular disease, depression, rheumatoid arthritis, and cancer [19]. Chaiklahan et al. [21] optimized the extraction of polysaccharides from Spirulina sp. It was found that the mostly significant operation conditions were temperature and solid to liquid ratio and time. The extract contains rhamnose and phenolic content [21]. Seaweed concentrates are used as supplementary soil conditioners that promote plant growth and improve crop yield [29]. An example product is Kelpak from Ecklonia maxima [29]. These products are used in very low doses and contain, for example, cytokinins and auxins that are plant growth regulators [29]. Seaweed extracts are particularly useful if applied on plants that are nutrient-stressed [29].

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1.6 Conclusions

Algae are a useful raw material for biobased economy, because their cells contain a vast array of useful compounds with high biological activity. Biomass of algae is certainly an underestimated resource. In the process of extraction it is possible to draw the valuable compounds closed in the algal cells. However, this should be carried out in such a way that the structure and thus the properties of the compounds are not destroyed and that the solvent used does not limit their use as safe components of products for plants, animals, and human.

References

There are many ways to implement the extraction process and this is thoroughly discussed in this book. In addition to developing extraction technology, it is very important to assess the utilitarian values of the extracts, which can be documented in application studies of extracts in real systems. Preparation of algal extracts represents a new approach in the preparation of natural products with a standardized composition, as compared with the biomass itself and certainly will be a future for algal industry. References 1. Shanab, S.M.M., Mostafa, S.S.M.,

2.

3.

4.

5.

6.

7.

8.

Shalaby, E.A., and Mahmoud, G.I. (2012) Aqueous extracts of microalgae exhibit antioxidant and anticancer activities. Asian Pac. J. Trop. Biomed., 2, 608–615. Spolaore, P., Joannis-Cassan, C., Duran, E., and Isambert, A. (2006) Commercial applications of microalgae. J. Biosci. Bioeng., 101, 87–96. Sharma, H.S., Shekhar, S., Lyons, G., McRoberts, C., McCall, D., Carmichael, E., Andrews, F., and McCormack, R. (2012) Brown seaweed species from Strangford Lough: compositional analyses of seaweed species and biostimulant formulations by rapid instrumental methods. J. Appl. Phycol., 24, 1141–1157. Borowitzka, M. (2011) Pharmaceuticals From Algae, Biotechnology, vol. 7, Encyclopedia of Life Support System. Gupta, S., Cox, S., Rajauria, G., Jaiswal, A.K., and Abu-Ghannam, N. (2012) Growth inhibition of common food spoilage and pathogenic microorganisms in the presence of brown seaweed extracts. Food Bioprocess Technol., 5, 1907–1916. Gupta, S. and Abu-Ghannam, N. (2011) Recent developments in the application of seaweeds or seaweed extracts as a means for enhancing the safety and quality attributes of foods. Innovative Food Sci. Emerging Technol., 12, 600–609. Stolz, P. and Obermayer, B. (2005) Manufacturing microalgae for skin care. Cosmet. Toilet., 120, 99–106. Ilbanez, E., Herrero, M., Mendiola, J.A., and Castro-Puyana, M. (2012) in Marine Bioactive Compounds: Sources,

9.

10.

11.

12.

13.

14.

15.

16.

17.

Characterization and Applications (ed M. Hayes), Springer, pp. 55–98. Borowitzka, M.A. (2013) High-value products from microalgae—their development and commercialization. J. Appl. Phycol., 25, 743–756. Plaza, M., Cifuentes, A., and Ibanez, E. (2008) In the search of new functional food ingredients from algae. Trends Food Sci. Technol., 19, 31–39. Balboa, E.M., Conde, E., Moure, A., Falqué, E., and Domínguez, H. (2013) In vitro antioxidant properties of crude extracts and compounds from brown algae. Food Chem., 138, 1764–1785. Alves, A., Sousa, R.A., and Reis, R.L. (2013) A practical perspective on ulvan extracted from green algae. J. Appl. Phycol., 25, 407–424. Herrero, M., Mendiola, J.A., Plaza, M., and Ibanez, E. (2013) in Advanced Biofuels and Bioproducts (ed. J.W. Lee), Springer, pp. 833–872. Craigie, J.S. (2011) Seaweed extract stimuli in plant science and agriculture. J. Appl. Phycol., 23, 371–393. Pulz, O. and Gross, W. (2004) Valuable products from biotechnology of microalgae Mini-Review. Appl. Microbiol. Biotechnol., 65, 635–648. Contreras-Porcia, L., Callejas, S., Thomas, D., Sordet, C., Pohnert, G., Contreras, A., Lafuente, A., Flores-Molina, M.R., and Correa, J.A. (2012) Seaweeds early development: detrimental effects of desiccation and attenuation by algal extracts. Planta, 235, 337–348. Lee, J.C., Hou, M.-F., Huang, H.-W., Chang, F.-R., Yeh, C.-C., Tang, J.-Y., and Chang, H.-W. (2013) Marine algal natural products with antioxidative,

11

12


18.

19.

20.

21.

22.

23.

24.

25.

anti-inflammatory, and anti-cancer properties. Cancer Cell Int., 13, 55–62. O’Sullivan, A.M., O’Callaghan, Y.C., O’Grady, M.N., Queguineur, B., Hanniffy, D., Troy, D.J., Kerry, J.P., and O’Brien, N.M. (2011) In vitro and cellular antioxidant activities of seaweed extracts prepared from five brown seaweeds harvested in spring from the west coast of Ireland. Food Chem., 126, 1064–1070. Kindleysides, S., Quek, S.-Y., and Miller, M.R. (2012) Inhibition of fish oil oxidation and the radical scavenging activity of New Zealand seaweed extracts. Food Chem., 133, 1624–1631. Onofrejova, L., Vasickova, J., Klejdus, B., Stratil, P., Misurcova, L., Kracmar, S., Kopecky, J., and Vacek, J. (2010) Bioactive phenols in algae: the application of pressurized-liquid and solid-phase extraction techniques. J. Pharm. Biomed. Anal., 51, 464–470. Chaiklahan, R., Chirasuwan, N., Triratana, P., Loha, V., Tia, S., and Bunnaga, B. (2013) Polysaccharide extraction from Spirulina sp. and its antioxidant capacity. Int. J. Biol. Macromol., 58, 73–78. Volland, S., Schaumlöffel, D., Dobritzsch, D., Krauss, G.-J., and Lütz-Meindl, U. (2013) Identification of phytochelatins in the cadmium-stressed conjugating green alga Micrasterias denticulata. Chemosphere, 91, 448–454. Farvin, K.H.S. and Jacobsen, C. (2013) Phenolic compounds and antioxidant activities of selected species of seaweeds from Danish coast. Food Chem., 138, 1670–1681. Rathore, S.S., Chaudhary, D.R., Boricha, G.N., Ghosh, A., Bhatt, B.P., Zodape, S.T., and Patolia, J.S. (2009) Effect of seaweed extract on the growth, yield and nutrient uptake of soybean (Glycine max) under rainfed conditions. S. Afr. J. Bot., 75, 351–355. Spinelli, F., Fiori, G., Noferini, M., Sprocatti, M., and Costa, G. (2010) A novel type of seaweed extract as a natural alternative to the use of iron chelates in strawberry production. Sci. Hortic., 125, 263–269.

26. Cutler, H.G. and Cutler, S.J. (2007) in

27.

28.

29.

Encyclopedia of Chemical Technology, vol. 13 (ed. K. Othmer), John Wiley & Sons, Inc., pp. 1–36. Sharma, H.S.S., Fleming, C., Selby, C., Rao, J.R., and Martin, T. (2014) Plant biostimulants: a review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. J. Appl. Phycol., 26, 465–490. Jannin, L., Arkoun, M., Etienne, P., Laîne, P., Goux, D., Garnica, M., Fuentes, M., San Francisco, S., Baigorri, R., Cruz, F., Houdusse, F., Garcia-Mina, J.-M., Yvin, J.-C., and Ourry, A. (2013) Brassica napus growth is promoted by Ascophyllum nodosum (L.) Le Jol. Seaweed extract: microarray analysis and physiological characterization of N, C, and S metabolisms. J. Plant Growth Regul., 32, 31–52. Papenfus, H.B., Kulkarni, M.G., Stirk, W.A., Finnie, J.F., and Van Staden, J. (2013) Effect of a commercial seaweed extract (Kelpak ) and polyamines on nutrient-deprived (N, P and K) okra seedlings. Sci. Hortic., 151, 142–146. Stirk, W.A. and Van Staden, J. (1997) Comparison of cytokinin- and auxinlike activity in some commercially used seaweed extracts. J. Appl. Phycol., 8, 503–508. El-Baky, H.H.A., El-Baza, F.K., and El Baroty, G.S. (2010) Enhancing antioxidant availability in wheat grains from plants grown under seawater stress in response to microalgae extract treatments. J. Sci. Food Agric., 90, 299–303. doi: 10.1002/jsfa.3815 Santoyo, S., Herrero, M., Senorans, F.J., Cifuentes, A., Ibanez, E., and Jaime, L. (2006) Functional characterization of pressurized liquid extracts of Spirulina platensis. Eur. Food Res. Technol., 224, 75–81. Heussner, A.H., Mazija, L., Fastner, J., and Dietrich, D.R. (2012) Toxin content and cytotoxicity of algal dietary supplements. Toxicol. Appl. Pharmacol., 265, 263–271. Cavallo, R.A., Acquaviva, M.I., Stabili, L., Cecere, E., Petrocelli, A., and

®

30.

31.

32.

33.

34.

References

Narracci, M. (2013) Antibacterial activity of marine macroalgae against fish pathogenic Vibrio species. Cent. Eur. J. Biol., 8, 646–653. 35. Capelli, B. and Cysewski, G.R. (2010) Potential health benefits of Spirulina microalgae. A review of the existing literature. Nutra Foods, 9, 19–26. 36. Annapurna, V.V., Deosthale, Y.G., and Bamji, M.S. (1991) Spirulina as a source of vitamin A. Plant Foods Hum. Nutr., 41, 125–134.

37. Horn, S.J., Aasen, I.M., and Ostgaard, K.

(2000) Ethanol production from seaweed extract. J. Ind. Microbiol. Biotechnol., 25, 249–254. 38. Cho, J.Y., Kwon, E.-H., Choi, J.S., Hong, S.Y., Shin, H.W., and Hong, Y.K. (2001) Antifouling activity of seaweed extracts on the green alga Enteromorpha prolifera and the mussel Mytilus edulis. J. Appl. Phycol., 13, 117–125.

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Part I Cultivation and Identification of Marine Algae

Marine Algae Extracts: Processes, Products, and Applications, First Edition. Edited by Se-Kwon Kim and Katarzyna Chojnacka. © 2015 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2015 by Wiley-VCH Verlag GmbH & Co. KGaA.

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2 Identification and Ecology of Macroalgae Species Existing in Poland Beata Messyasz, Marta Pikosz, Grzegorz Schroeder, Bogusława Łe˛ska, and Joanna Fabrowska

2.1 Introduction

Algae are most common organisms in aquatic environment and a very diverse group in terms of ecological, taxonomic, morphological, and biochemical aspects [1–5]. Microscopic algae float freely in water and form plankton, which plays an important role in maintaining the balance of the aquatic habitat [6]. Macroscopic algae exhibit complex degrees of organization of thalli. Their main representatives are marine red algae (Rhodophyta), brown algae (Phaeophyta), and green algae (Chlorophyta), whose names are derived from the characteristic pigments phycoerythrin, fucoxanthin, and chlorophyll, respectively. The thalli of these algae, depending on the species, can reach a size of a few microns up to several meters. In the case of large marine thalli leaf-like (phylloid), stem-like (cauloid), and rootslike (rhizoids) forms can be found [6, 7]. On the basis of a wide morphological diversity and biochemical characteristics algae have traditionally been classified into several taxonomic groups (phyla). Organizing algae according to the principles of the phylogenetic system is still rather difficult. The first system of algae classification based on the theory of parallel development of monophyletic groups of algae was derived from flagellates and then included various degrees of morphological organization. According to this compilation, highly organized filamentous and pseudoparenchymatous forms arose from primitive unicellular flagellate algal cells [2, 3, 7–9]. In this system, representatives of marine and freshwater macroalgae are included, such as green algae (e.g., Ulva, Cladophora), red algae (e.g., Batrachospermum, Porphyra, Polysiphonia, Furcellaria), brown algae (e.g., Fucus, Laminaria), cyanobacteria (Tolypothrix, Scytonema, Nostoc), and xanthophyceans (e.g., Vaucheria, Tribonema) [10, 11]. Some of these genera are found in freshwater ecosystems in abundant quantities (Figure 2.1). Interestingly, often their presence in the aquatic reservoirs is generally defined as “filamentous green algae” by the researchers without identifying the species structure of such mats. Significantly, this makes it difficult to characterize the ecology of individual species, and a comparison with Marine Algae Extracts: Processes, Products, and Applications, First Edition. Edited by Se-Kwon Kim and Katarzyna Chojnacka. © 2015 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2015 by Wiley-VCH Verlag GmbH & Co. KGaA.

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2 Identification and Ecology of Macroalgae Species Existing in Poland

Chlorophyta

Phylum Ulvophyceae

Class Ulotrichales

Ulvales

Ulotrichaceae

Ulvalceae

Ulothrix

Ulva

Chlorophyceae Microsporales

Oedogoniales

Pitophoraceae

Microsporaceae

Oedogoniaceae

Aegagriopila

Microspora

Oedogonium

Cladophorales

Order Familly Genus

Cladophoraceae Cladophora

Rhizoclonium

Ochrophyta

Phylum

Xantophyceae

Class Order

Vaucheriales

Familly

Vaucheriaceae

Genus

Vaucheria

Tribonematales Tribonemataceae Tribonema

Figure 2.1 Systematic diagram of most often recorded filamentous algae from water ecosystems in Poland.

neighboring countries or regions is also not possible as it will require a thorough verification of such incompletely reported occurrences. In the marine environment, in contrast to the freshwater, the occurrence of macroscopic algae is influenced by the availability of light. Zonation takes place, where red algae can develop in the lowest part of the water column. Likewise, mass macroalgae developments are also found in inland water source; however, the scale of such blooms is lower than it is in marine water because of the smaller size of the reservoirs. These are mainly representatives of green algae and to a lesser extent of xanthophyceans (Figure 2.2). Cyanobacteria, irrespective of the type of water ecosystem, are only an accompanied group in the macroalgal associations as they do not create a large surface mat by themselves. Their filamentous forms can grow as a thin mat over stones, break away and become free-floating (Stigonema), form dark tufted mats (Scytonema), or tangle among submerged vegetation (Tolypothrix). On the contrary, red and brown algae are predominantly marine. Only few species are found in freshwaters (e.g., Batrachospermum, Lemanea). There are many others that have not been fully studied and their ecological characteristics are still not well described. Macroscopic green algae and xanthophyceans although usually free-floating thalli forming dark green patches may be attached when they are young and before breaking free. The speed of the growth of macroscopic algae biomass is influenced by environmental conditions that vary according to the season. These algae in terms of longevity are the annual forms. Therefore, the variability of macrogreen algae mats will concern the composition and species diversity, the structure of patches (loose or dense) as well as the occupied area. However, in each case the rapid algal growth permits their rapid settling on the available

2.1

(a)

(b)

Introduction

(c)

(d)

(e)

(f)

(g)

(h)

(i)

Figure 2.2 Massive development of filamentous green algae forming mats from Wielkopolska region (Poland): (a) Ulva intestinalis in Nielba river (Photo by M. Koperski.); (b) Oedogonium in the Konojad pond; (c) filaments of Cladophora glomerata in Lake Durowskie; (d, e) Cladophora glomerata

in Lake Oporzyn; (f ) Cladophora fracta in the Malta Reservoir; (g) long filaments of Cladophora glomerata in the Mogilnica river; (h) Cladophora rivularis in the Konojad pond; and (i) Zygnemataceae in the artificial pond ´ in Poznan´ (Photo by J. Rosinska).

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substrates. Our long-term studies have shown that successional stages are less predictable in freshwater ecosystems than in marine ones although one clearly can distinguish the spring phase with filamentous ephemerals such as Ulothrix or Tribonema and the summer phase with a dense carpet/mats of Cladophora, Oedogonium, or Ulva [12].

2.2 Collection of Macroalgal Thalli and Culture Conditions

Macroalgae collection is dependent on the habitat in which these organisms occur. Because of the considerable depth of water where these are available a collector often needs a boat. For the habitat characterization, the basic physiochemical parameters of the water (temperature, conduction, concentration of oxygen and Cl− as well as the pH) at the sites of macroalgae thalli are measured with the use of the YSI Professional Plus hand-held multiparameter meter. Thalli samples are collected manually from the middle of the mat, which is formed by the macroalgae at the sampling site. When a macroalgal mat is not floating on the water surface, thalli samples are collected under water, often by gathering individual creepings at the bottom or tangled in aquatic vegetation. It is recommended that about 500 g of algal thalli is collected, which are rinsed five times with water from a given site. The thalli are put in a plastic container and transported in a fridge (at 4 ∘ C) to the laboratory, where they are rinsed again a couple of times with distilled water in order to remove any algae, vascular plants (lemnids), sand, or snails stuck to them. Next, the purified thalli belonging to one genus or species are divided into smaller portions: (i) 10 g is used for the microscopic analysis and morphometric measurements of both thalli and cells, (ii) 20 g is used to perform the herbarium specimens, (iii) another 30 g of the sample is preserved (including 10 g of the material frozen in the temperature of −10 ∘ C, 10 g preserved with 4% formalin solution, and 10 g preserved with Lugol’s solution; put into 100 ml plastic containers), and (iv) to analyze the chemical composition, 20 g of thalli is dried for 30 min on a cellulose filter paper at room temperature and then for 2 h at 105 ∘ C. The obtained dry mass is stored in 100 ml plastic containers. The remaining 400 g of the collected sample is placed in an 10 l aquarium with water filtered from the habitat or the Wang medium and next deposited into phytotrons (at 250 μmol photons m−2 sek−1 , period 12 : 12, temperature 21 ∘ C) to conduct macrocultures in open or closed systems [13]. In order to obtain high quality raw materials for the production of food products and cosmetics, cultures of algae are treated more frequently under specially defined conditions to increase the biomass production. Open cultures are mainly related to algae culture on a large scale or in cases where experimental sets occupy a large surface. The main element of such a culture is a container with water for the growth of algae. Other components are subject to various modifications, depending on the needs and purpose of the experiment. Cultures focused on obtaining the highest algal biomass are built from containers of large capacity and equipped only

2.3

Macroalgae Forming a Large Biomass in Inland Waters of Poland

with monitoring devices and aerator [14]. Sets for breeding algae with other organisms (snails, shrimps) are more complicated. The latter are chosen to examine the mutual ecological relations [15, 16]. Such sets consist of several smaller containers connected by canals with water circulation. More complex sets of open culture are equipped with pipes supplying water enriched with nutrients, the heater controlling the water temperature, devices for simulating the movements of water or artificial light sources [17]. And, the thalli of algae (instead of freely floating on the water) are deposited on the special nets or other supports [18]. Phytotron chambers are used, which permit the cultivation of algae under certain simulated conditions. In these chambers several environmental factors can be modeled, such as (i) air temperature (set by the heating and cooling systems, maintaining the temperature regardless of the surroundings), (ii) circulation and humidity (provided by a system of fans and filters), and (iii) intensity and color of light (system of lamps and day and night cycle). Embedded microprocessors allow the automatization of processes and controlling proper operational parameters. Tunable components of the culture medium are nitrogen, phosphorus, pH buffers, salinity (minimum 30 ppt), and optionally trace metals and vitamins as defined by the medium recipe. In the case of marine algae culture, to obtain the required amount of biomass, the physicochemical properties of the marine water from the site from which a specific macroscopic algae species are harvested are analyzed; then an appropriate amount of chemicals is added , which leads to certain nutrient and trace element concentrations. A key factor that determines the success of the cultured freshwater forms of macroalgae is to select the appropriate media. On the basis of the observations of the concentration of nutrients for the macroalgae, it has been found that Wang’s culture medium or the Benecke medium (with some modifications) [13] are most suitable. The relatively high contents of N and P present in these media are similar to those existing in a eutrophic reservoir habitat – preferred by mat-forming algae. NaCl is added to the medium, for example, in the case of the culture of freshwater forms of Ulva (preferences are different for individual species) to complete chloride ions or the addition of trace elements mixture in the cultivation of other filamentous green algae species. A very important aspect of culture preparation is the identification of the species that were collected for testing. Species of the genus Cladophora, Ulva differ substantially in terms of levels of certain nutrient preference. It is therefore necessary to adjust the amount of the nutrient element in the medium to the requirements of the identified macroalgae.

2.3 Macroalgae Forming a Large Biomass in Inland Waters of Poland

This chapter relates to macroalgae living in the freshwaters of Poland. However, some of these algae such as Ulva spp. (Enteromorpha spp.), Cladophora spp., and Vaucheria spp. are represented also in the marine waters of the Polish coast where they cover the stone bottom. In some places stoneworts are also rooted

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(Chara spp.). On the contrary, brown alga Fucus vesiculosus L., which is specified as the most characteristic plant of the Baltic Sea, has become extinct on the Polish coast completely [10, 11, 19, 20]. Thirty years ago, it occurred abundantly at the stony bottom near the cliffs and in the Puck Bay together with the aquatic plant Zostera marina L. and red alga Furcellaria lumbricalis (Hud.) Lam., creating an association of underwater meadows with an incredible biodiversity. Nowadays, fragments of this brown alga are more often found on our beaches as detached by water from the other parts of the Baltic Sea. A very common filamentous brown alga in the southern Baltic now is Pilayella littoralis (L.) Kjell. [11, 21]. Its delicate and thin thalli are strongly and diversely branched. It creates dense bushes of yellow brown color reaching several centimeters in length. During the summer it grows strongly. Moreover, this species has a tendency to spread rapidly along the Polish coast. In the case of tubular forms of marine green algae from the genus Ulva (Enteromorpha), Ulva compressa L. and Ulva plumosa Hud. are present widely while less represented (and in isolated locations) are Ulva clathrata (Roth) Ag., Ulva linza L., Ulva prolifera O.F. Müller, and Ulva torta (Mert.) Trev. [22–25]. The pale green thallus of U. compressa is shaped like a flattened tube with delicate and very thin cell walls. Length of thalli can reach dozens of centimeters and grow up to 2 cm in width. They are often distended in the form of bubbles because of the air penetrating the thalli. Its thallus narrows in intervals, from which new branches extend. This green alga strongly attaches via disc-like basal cell to solid substrates. Waves do not cut these, but only sways them. However, U. plumosa, which is very common in the Baltic Sea, is heavily branched and forms a bundle of long, delicate thalli in an intensive and luscious green color. In the case of a soft bottom it is attached to shells or pebbles. Its filamentous thalli reach more than 30 cm in length. In addition, in marine ecosystems, despite the most common filamentous ones like Cladophora glomerata (L.) Kütz., there are also other species, such as Cladophora fracta (O.F. Müller ex Vahl) Kütz. (Figure 2.2f ), Cladophora albida (Nees) Kütz., Cladophora sericea (Hud.) Kütz. [11, 22, 23, 26]. A very interesting species is Cladophora rupestris (L.) Kütz., which has a characteristically branched thallus. From the apical part of the filament protrude a few branches, from each of them arise again three to four branches forming a kind of brushes. It is easy to identify because of the strong ramification thalli of this alga and its rigidity. Such light green and fluffy bushes can reach about 15 cm in height. It tolerates a wide range of temperatures and thus is a perennial species. However, it does not grow during winter and is not fruitful. On the basis of the findings from our long-term studies and all available literature, the characteristics of the biology and ecology of select macroalgae taxa are described below. For each species the same pattern of presentation is chosen, including macro- and microscopic appearance, habitat preferences, place of occurrence, and characteristics of the communities in which they were recorded.

2.3


ULOTHRIX VARIABILIS Kützing 1849 (Figure 2.3m,q) (Chlorophyta, Ulvophyceae). 10 μm

(a)

(d)

(b)

(c)

(e)

(f)

(g)

(i)

(h)

(j)

(l)

(k)

(n)

Figure 2.3 Morphology of (a) highly branched thalli of Cladophora glomerata; (b) numbers of nucleus in C. glomerata cell after acetocarmine staining; (c) branched Cladophora rivularis; (d) ball form of Aegagropila linnaei; (e) filament of A. linnaei with characteristic opposite and subterminal branches; (f ) unbranched filament of Rhizoclonium sp.; (g) filament with H-shape cell membrane of Microspora sp.; (h) Ulva intestinalis thallus with proliferation in the lower

(o)

(m)

(p)

(q)

part; (i) thalli of Ulva flexuosa subsp. pilifera; (j) cells of Ulva sp.; (k) unbranched filament of Oedogonium capillare with pyrenoids; (l) Oedogonium sp. with apical cell; (m) Ulothrix variabilis with single, parietal, girdle-shaped chloroplast; (n) coenocystic, hollow tube of Vaucheria sp.; (o) antheridium of Vaucheria sp.; (p) Tribonema aequale with H-shape pieces; and (q) filaments of Tribonema and Ulothrix.

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Synonyms: No data. ̇ (Polish) [27]. Common names: wste˛znica Macroscopic appearance: Unbranched thin filaments. Microscopic appearance: Cylindrical cells 4.5–7 μm thick, 0.5–1.5 times as long as broad, 1 pyrenoid [28]. Cells cylindrical, 8–14 μm long, 6–9 μm wide, end of cells rounded, single pyrenoid, containing numerous starch grains and central nucleus [29]. Cylindrical cells 4.5–7 μm width and 0.5–1.5 times longer [27]. Square shape cells (width: 2.5–6 μm length: 5–6 μm) with two chloroplast located on the sidewall, rounded apical cell (own research). Habitat: Reported as a terrestrial species, it may be attached or free-floating on the water. The alga has a cosmopolitan distribution mainly in stagnant, flowing waters particularly at cooler times of the year. Light (grassy) green, forming delicate watt. It can also be attached to submerged stones or wood. Communities: In the littoral zone Ulothrix forming mats with Oedogonium, Spirogyra, Zygnema, and Mougeotia [30]. Often included in phytoplankton community, were noted with Tribonema aequale and T. vulgare [12]. Distribution: This species is found in a variety of small pools and shallow water bodies as well as in soil but in small amounts. Massive occurrence was noted in April in midfield pond in Konojad village [12]. Ulothrix species were noted on the Spitsbergen in terrestrial ecosystems [31]. Remarks: About 30 species of Ulothrix genera are known. ULVA FLEXUOSA SUBSP. PILIFERA (Kütz.) Bliding 1963 (Figure 2.3i,j) (Chlorophyta, Ulvophyceae). Synonym: Enteromorpha pilifera Kützing. Common names: błonica oszczepowata, watka oszczepowata (Polish) [32]. Macroscopic appearance: Monostromatic tubular thalli long, 15–30 × 1–3 cm [33] and according to Rybak and Messyasz [32] 15–41 × 0.4–4.2 cm. Thalli of macroalgae can reach length up to 1 m [27]. Thalli with little proliferation or without [33, 34]. This species has an entero-folding type of construction and can be found in the submerged form and free-floating mats. Microscopic appearance: Cells have the square or rectangular shape with rounded edges, 14.4–24 × 9.9–16.6 μm [33]. Cell size 11.6–21.1 × 7.8–17.4 μm with 1–3 diameters of pyrenoids 2.1–2.6 μm [32]. Habitat: U. flexuosa subsp. pilifera cosmopolitan euhalinity species, wide distribution in marine, freshwater, and brackish environments throughout the world except arctic ecosystems [35–37]. Noted in water: pH-7.98, conductivity682 μS cm−1 , TDS (Total Dissolved Solids)-443 mg l−1 ; average concentration of NO3 − -0.05, NH3 − -0.54, NH4 + -0.57, P-PO4 -0.023, P2 O5 -0.052, PO4 3− -0.07, Cl-107.5, NaCl-177.37 mg l−1 , and total Fe-0.02 mg l−1 [32]. Communities: Among macrophytes, Phragmites australis, Myriophyllum spicatum, Glyceria maxima, Polygonum amphibium, Lycopus europaeus, and Alisma plantago-aquatica [32]. Ulva could be woven into filamentous green algae from the genera Oedogonium (Chlorophyta). Distribution: In Poland this taxon was recorded in the channel ion Mie˛dzyodrze, Szczecin [34]; in the ponds in Piotrowice and in Kuciny, Łód´z [33],

2.3


in a fishpond in Arturówko, Łód´z (M. Sitkowska, 2008 – unpublished); in the Wielkopolska region, for example, Maltański Reservoir [32]; and in the retention tank in the northern part of Poznań [38]. Abundant in the different ecosystems of Poland. Remarks: U. flexuosa subsp. pilifera is a dominating taxon from the genera Ulva in freshwater ecosystems of Europe [5]. ULVA INTESTINALIS L. 1753 (Figures 2.2a and 2.3h) (Chlorophyta, Ulvophyceae). Synonyms: Conferva intestinalis (Linnaeus) Roth 1797, Ulva enteromorpha var. intestinalis (Linnaeus) Le Jolis 1863, Enteromorpha compressa var. intestinalis (Linnaeus) Hamel 1931, Enteromorpha intestinalis f. maxima J.Agardh 1883, E. intestinalis (L.) Ness. Common names: ta´sma kiszkowata (Polish), Gut Weed (English) [39], Darmtang (Germany) [40], Tarmalg (Swedish) [41]. Macroscopic appearance: Freshwater thalli are tubular, wrinkled, with numerous prolifications. Young thalli are attached to the substrate, whereas mature float on the water surface. Its thallus is tapered at the scutellum that attaches to the substrate, and further expands and stays the cylindrical up to the apical part. Microscopic appearance: Thalli cell wall consists of a single layer of cells, which may take the form of an oval or even rectangular shape, having a diameter of 10–25 μm and a thickness of 16–18 μm; chloroplasts are thin and arranged parietally [42]. Habitat: Frequently found in the coastal zone of seas and oceans, estuary waters [43, 44], U. intestinalis is a typical euryhalin species [45], therefore its tolerance to salting waters [18]. U. intestinalis grows best with a salinity around 24‰ but is also listed in areas with lower values of this parameter [46]. Communities: Often occurs with filamentous green algae, mainly C. glomerata. Distribution: Widespread, massively growing on empty shells, larger stones, and port breakwaters. In the Polish part of the Baltic Sea U. intestinalis was found ´ in the littoral zone of water bodies in the Władysławowo [23, 47], Swinouj´ scie, Kołobrzeg, Łeba, Mielno [22], in the Gdańsk Bay [21, 23, 24, 48, 49], and in the Puck Bay [22–25]. The inland site of U. intestinalis in Poland was found in Mie˛kinia and Duszniki Zdrój in the Lower Silesia [50], in the ditch near Ciechocinek [51], in lakes near Inowrocław and in ponds and river – Kołobrzeg [52], lakes near Da˛bskie, Miedwie and rivers in Płonia [34], clay pit near Warszawa ̇ [53], Lake Zarnowieckie [54], Lake Dymer near Olsztyn [55], Lake Laskownickie, the Nielba river [56], the pond Biskupice near Lublin [57]. This species was recorded also in the littoral of lakes supplied periodically by marine waters such as Lake Gardno and Lake Łebsko. Remarks: U. intestinalis (E. intestinalis) is the most common Ulva species in Poland. CLADOPHORA GLOMERATA (Linnaeus) Kützing 1843 (Figures 2.2c–e,g and 2.3a,b) (Chlorophyta, Cladophoraceae). Synonyms: Conferva glomerata Linnaeus 1973.

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Common names: gałe˛zatka (Polish) [27], grönslick (Swedish) [41], blanket weed (English). Macroscopic appearance: Siphonocladous thalli with multinucleated cells. Species with high variability of morphometric traits, often confused with Rhizoclonium. Its thalli can reach up to several meters in length. Young, short thalli are usually highly branched and older have fewer and longer branches occurring mainly in the lower part. Species is characterized by pseudodichotomous type of branching. Forms may be either attached to various substrates using rhizoids or be loosely floating to form the mat. The color is usually dark green. Because of the presence of numerous epiphytes (diatom, green algae, and cyanobacteria) and incrusted on the surface of the cell wall its filaments are rough to the touch. Microscopic appearance: Double-layer cell wall. Its apical cell width is (19–24)−(58–91) μm, while the main filament is 150 μm [27]. Cell size range in the main axis cylindrical cells is 90–100 μm in diameter and 160–240 μm long; branches 40–70 μm in diameter and 150–270 μm long. Apical cells 20–40 μm in diameter and 140–160 μm long [58]. The width of apical cell is 37–54 μm, the main filament 100 μm, pyrenoids 3.0–6.4 μm in diameter, the nucleus diameter 4.6–8.5 μm, thickness of the cell wall 3–4 μm (own study). Habitat: Cosmopolitan species, common in marine and littoral ecosystems (estuaries), saline, and freshwater. Commonly found in running waters and lakes/ponds. So far not reported in polar waters. Found as the most frequent species of the genus Cladophora. Habitat factors such as the water temperature (17–29 ∘ C), a neutral pH (pH > 8), the availability of light (mean 3000 LUX), and the content of nutrients have a huge impact on the presence of this macroalga. Furthermore, it prefers clear waters with color values not above 30 mg Pt/l [59]; however, in the Mogilnica river, these values are about 40. Communities: It can create single-species and multispecies mat in which is the core component. C. glomerata occurs with taxa belonging to the genus Oedogonium, Stigeoclonium, Microspora, Mougeotia, Spirogyra [60, own study]. In the mountain streams [61], as in the lowland river [62], this species was present at the site with red alga Hildenbrandia rivularis. In the Baltic Sea it is noted with Enteromorpha compressa, E. flexuosa subsp. flexuosa, and E. linza [25]. In the littoral zone this green alga is present in the vicinity of macrophytes. Distribution: Common in Polish Baltic Sea: Gdańsk Gulf [20, 22, 23], east coast of the Pomorska Gulf [26], Sopot (own study); mountain rivers, and streams: Skawa [63], Lubogosz [61]; lowland rivers: Nielba, Wełna, Mogilnica, Samica Ste˛szewska (own study); lakes: Ro´s [60], Durowskie, Oporzyn, Zbiornik Maltański, and different types of small water bodies (own study). Certain findings of this species are from protected regions such as the Woliński National Park, the Słowiński National Park, and the Wielkopolski National Park. Remarks: It occurs particularly often in highly morphologically transformed and eutrophic locations. Shows a significant increase in biomass concentration (can three times increase its biomass per day). It is possible to use it as a biomarker of water pollution by heavy metals.

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CLADOPHORA RIVULARIS (L.) Van Hoek 1963 (Figures 2.2h and 2.3c) (Chlorophyta, Cladophoraceae). Synonyms: C. fracta var. rivularis (L.) Brand, C. crispate (Roth) Kützing, C. glomerata var. fluitans (Kütz.) Rabenh., C. oligoclona (Kütz.) Kützing, C. insignis (C. Ag.) Kützing. Common names: No data. Macroscopic appearance: Poorly branched, long, intertwining filaments, and often with variable diameter. These species may be mistaken with Rhizoclonium spp., Chaetomorpha linum. Color usually light green. Forming dense mat on the water surface. Microscopic appearance: Diameter < 30 μm (18–25 μm) cell in the filament cylindrical, 50–100 μm in diameter and 300–440 μm long; cell wall thin layer, chloroplast periphery, reticulate, numerous discoid pyrenoids, and conspicuous [58]. Morphological variability: the main axis continued to grow with cells being half the diameter of the origin size. Cell 38–76 μm in diameter and 170–387 μm long, cell wall 3–7 μm; rounded apical cell (W/L 35/320 μm), number of nucleus 4.5–8.0 μm, and pyrenoids 10 μm in diameter. Habitat: C. rivularis is typical for stagnant and turbulent fresh water. Development in eutrophic condition, pH > 7, conductivity ∼1000 μS cm−1 , total dissolved substance 440 mg l−1 , and chlorides 550 mg l−1 . Communities: Mixed with C. glomerata var. glomerata among Typha angustifolia [64]. In streams coexisted with other Cladophora species [58]. In midfield pond dominant with Oedogonium sp. and formed algal-cyanobacterial metaphyton [12]. Remarks: It was common in Silesia Region – Strzelin [27]. Forming dense mat in Konojad Pond (Wielkopolska Region). RHIZOCLONIUM Kützing 1843 (Figure 2.3f ) (Chlorophyta, Cladophoraceae). Synonym: No data. Common names: Ryzoklonium (Polish) [27], Grönkrull (Swedish) [41]. Macroscopic appearance: A single filament without or only with a short branch. Rhizoclonium species are difficult to differentiate from some Cladophora species with rare branches. Microscopic appearance: Usually only one nucleus in the cell, sometimes four. Chloroplast reticulate, parietal, with many pyrenoids, which are densely packed with starch [7]. Width of cell is usually equal across the filaments (10–50 μm). Habitat: Common in hard and shallow waters. Rhizoclonium cosmopolitan in fresh, brackish, and marine waters, often growing entangled with other algae or forming a dense mat [7]. Communities: R. fontanum coexisting with Microspora fontinalis in cold, slowrunning waters [27]. Rhizoclonium cf. hieroglophylicum were noted with U. variabilis and Microspora floccosa in lakes near peat bogs [65].

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Distribution: In drainage ditches near Olsztyn [66], Rhizoclonium cf. hieroglophylicum in a small dystrophic reservoir – Lake Ku´zniczek near Piła [65]. Representatives of this genus were recently found in the “heated” lakes near Konin with regular discharges of heated waters from Konin and Pa˛tnów power plants. Remarks: There is more than 70 species of Rhizoclonium. AEGAGROPILA LINNAEI Kützing (Figure 2.3d and e) (Chlorophyta, Pithophoraceae). Synonyms: Cladophora aegagropila var. linnaei (Kützing) Rabenhorst 1868, Cladophora sauteri (Nees ex Kützing) Kützing, Aegagropila profunda (Brand) Heering 1921. Common names: Lake balls, Cladophora balls (English) [64], Marimo (Japanese) [67], gałe˛zatka kulista (Polish) [27], Getraggsalg (Swedish) [41]. Macroscopic appearance: Rather simple morphology, growth form as characteristic balls. Attached or unattached mats/balls floating on the sediment and filaments are yellowish to dark green. Microscopic appearance: A. linnaei characterized by subterminal, lateral, opposite, and serial insertion of dense branches [64]. Irregular cell shape and variable cell dimension: apical cells 30–70 μm width, main axis cells 125–200 μm width [27]; apical cell rounded 45–71 μm width and main axis cells 136–363 μm width (own study). Habitat: Cosmopolitan. Freshwater and brackish water [64], particularly well developed in eutrophic freshwater lakes [27]. A. linnaei is generally a rare species that has only 283 recorded [68]. It can occur in several different growth forms, depending on environmental conditions [69]. Communities: Because of the ability of “lake balls” to move/roll, the presence of other macroalgae is not reported. Mathiesen and Mathiesen [70] described the Aegagropiletum benthonicum association composed of C. aegagropila (= A. linnaei), attached or forming loose-lying balls from Gulf of Bothnia. Distribution: Until now we know six locations of A. linnaei in Poland: Gulf of Puck, Jantar, Rewa, Sopot, Lake Miedwie near Szczecin, and Lake Tatarak near Legnica, Lake Wierzbnickie near My´slibórz [23, 71]. Remarks: Common used in aquarium as a decorative plant. MICROSPORA Thuret, 1850 (Figure 2.3g) (Chlorophyta, Chlorophyceae). Synonym: No data. Common names: Hantelalger (Swedish) [41]. Macroscopic appearance: Unbranched filamentous green algae with a holdfast for substratum attachment. Usually dark green. Microscopic appearance: H-shape cell membrane, thus may be mistaken with Tribonema species. In the cylindrical cell (5–25 μm diameter) pyrenoid is absent and only one nucleus is present, with characteristic netlike chloroplast filling all cells. Its cells are from one to three times as long as broad.

2.3


Habitat: Cosmopolitan freshwater species, usually abundant in small water bodies. Noted from ponds, rivers, ditch, peat bog, and the water pool [27]. Microspora communities may form watt on the water surface. Communities: Microspora spp. were noted with C. glomerata [72]. Distribution: In the Kamienna river [72]. Findings of this species are from lowland rivers in the Kujawy and Wielkopolska regions. Remarks: Frequent at cooler times of the year. OEDOGONIUM CAPILLARE Kützing ex Hirn 1900 (Figures 2.2b and 2.3k, l) (Chlorophyta, Oedogoniaceae). Synonyms: Conferva capillaries Linnaeus 1753, Oe. regulare Vaupell 1861, Oe. stagnale Kützing, Wittrock et Nordstedt 1883. Common names: No data. Macroscopic appearance: Young organisms usually attached to substratum such as macrophytes or stones by holdfast – specially adapted cell, in the later stage of development form free-floating mat. Microscopic appearance: Filaments of O. capillare are multicellular and unbranched with characteristic cap cells, cylindrical oogonium, and reticulate multipyrenoid chloroplast. Cylindrical cells 27–56 μm width and 30–102 μm long with 6, 8, or 10 pyrenoids [73]. Width of oogonium 45–75 μm and antheridium 30–48 μm [74]. Habitat: They are common in freshwater ecosystems and prefer growing in stagnant waters such as small ponds, pools, roadside ditches, marshes, old river bogs, lakes, reservoirs, and rivers [75–78]; however, the most numbers of Oedogonium taxon were noted in small water bodies and prefer habitats with high insolation. Physicochemical parameters: pH (7.6–9.5), electrolytic conductivity (635–663 μS cm−1 ), water temperature (21.1–25.0 ∘ C), TDS (442–455 mg l−1 ), and oxygen concentration (0.64–0.67 mg l−1 ) [73]. Communities: Oedogonium mostly form multialgal mats and might be associated with Spirogyra, Rhizoclonium [79]. O. capillare in small midfield pond (Wielkopolska Region) occurring massively in association with Microspora sp., Oedocladium sp., and Lyngbya hieronymusii [73]. Distribution: In Poland, this taxon was recorded from 1860 in the pool and ditch in Strzelin (near Wrocław) where appeared from June to October and from Domaszczyn [80]. T. Mrozińska-Webb (unpublished) observed a presence of this taxon of Oedogonium in the reservoir in Koniecwałd (Malbork) and from Upper Silesia in the river Przemsza in February 1978 in water at 6 ∘ C and 8 pH [77]. A last site of the O. capillare was located in the Wielkopolska province, in the Konojad village [73]. Remarks: Oedogonium genera includes 534 species [81]. Species identification is impossible without the presence of reproductive structures. VAUCHERIA De Candolle 1801 (Figure 2.3n,o) (Xanotphyta, Xantophyceae). Synonym: No data. Common names: prostnica (Polish), Slangalger (Swedish) [41], water felt (English).

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Macroscopic appearance: Multinucleate, coenocytic, branched filamentous yellow-green algae. Siphonous organization is uniaxial. Oogonium is round or oval, has a wall with a wide pore when mature, and is cut off from the stalk or main filament by a crosswall. Antheridium cut off by a crosswall from the stalk or main filament. Microscopic appearance: Cylindrical filaments 20–140 μm wide with irregular branching. There are many discoid chloroplasts with or without pyrenoids. The oogonia (female structure) and antheridia (male structure) form as lateral protuberances. Diagnostic features: vegetative filaments, oogonium, antheridium, oospora. Habitat: Vaucheria species prefer clear water from oligosaprobic to betamezosaprobic zone [82], cosmopolitan, widespread in freshwater and brackish. High temperature and poor oxygenation limited their development. Communities: Vaucheria were usually found in the company of filamentous green algae such as C. glomerata var. glomerata, Ulothrix tenuissima, Tetraspora sp., and xanthophycean Tribonema viride [83] and also grew in masses with Mougeotia sp. [84]. Distribution: Widespread in shallow freshwaters as well as in salt marshes and brackish waters. In Poland reported as very common in running waters, fishponds, and wet oils near Kraków [83]. From fishponds in Ochaby [85], near Łód´z [86], in Widawka river [87], salt flats near Łe˛czyca [88], in Czarna Przemsza river [89], from canals Giczno, Okre˛t, draining ditch in Młogoszyn, rivers Zalewska, Pisia, Grabia, Kłodnica, Bierawka, from dam in Walewice, and from southern Kujawy ´ [90], the small shallow pond in Olszyna reserve – Warsaw, Zródła Nałe˛czowianki springs, on moist soil in Botanical Garden of Warsaw University and from ditch in lower basin of Biebrza River [91], drainage ditch in Owczary reserve [84]. Remarks: Occurs in a wide range of fertility in morphometric differentiated water reservoirs. The various species of this genera are identified mainly by their reproductive structure. Precisely 20 species from Vaucheria genus are marine or brackish. TRIBONEMA AEQUALE Pascher 1925 (Figure 2.3p,q) (Ochrophyta, Xantophyceae). Synonyms: Conferva bombycina var. aequalis Kützing. Common names: No data. Macroscopic appearance: Filamentous, unbranched yellow-green algae. Slim in touch and green color. Microscopic appearance: Tribonema have H-shaped bipartite walls, usually long cells, elongate-cylindrical, cellulose cell wall, not coenocytic. Habitat: Common in freshwater ecosystems, especially those rich in organic and humic materials, but found to be rare. They may be included in the epiphytic organisms attached to a substrate as aquatic plants [74]. Communities: Small quantities of T. aequale filaments tangled in Batrachospermum moniliforme thalli [92].

2.4

Ecology Aspects of Freshwater Macroscopic Algae

Distribution: In old river bed [92], fishpond in Ochaby [74], small midfield pond (own research). Tribonema species were noted on the Spitsbergen in terrestrial ecosystems [31]. This species has been found in several different ponds in the Wielkopolska area, for example, in forest ponds on the meteorites origin in the “Meteorite Morasko” reserve. Remarks: Tribonema may grow in the dark utilizing glucose [93].

2.4 Ecology Aspects of Freshwater Macroscopic Algae

The occurrence of some green algae species is ephemeral; however, they appear mostly in eutrophic waters with high NaCl concentration of anthropogenic origin being present [94]. Intensive growth of these algae affects the physical–chemical properties of freshwater ecosystems, as they form extensively wide mats that freely float on the water surface. They are ubiquitous and widely distributed in aquatic reservoirs and occur in a very wide variety of habitats, for example, oceans, marshes and brackish waters, lakes, rivers, small freshwater bodies. Many species have a preference for alkaline, eutrophic waters, where they can appear as large biomass. On the contrary, macroscopic green algae and xanthophyceans are absent from oligotrophic locations [56]. It is likely that each species needs for its development a specific set of environmental conditions. Freshwater macroscopic green algae and xanthophyceans are primarily a component of the phytobenthos community in the littoral zone. They may also be found as a component of the plankton community where their survival in the euphotic zone is dependent on the availability of nutrients and its residence time. Algae from the genus Ulva, Cladophora, or Oedogonium have the ability to create very intense algal blooms. They are able to build vast mats floating on the water surface, covering large areas of water bodies and watercourses [95]. However, the reasons for the formation and development of such phenomena are still unclear because different species of algae prefer slightly different habitat conditions. Environmental conditions that determine the existence of these algae in freshwaters are primarily light (essential for autotrophic organisms), temperature, water movement, and pH as well as relevant concentrations of nutrients [3, 8]. Light is an important habitat factor, easily accessible to all species of macroalgae because they form macroscopic filaments or tubular thalli floating on the water surface. However, in the case of temperature, previous observations indicate that these algae prosper best at moderate to high temperatures. Many species of Tribonema, Ulothrix, Vaucheria, and Rhizoclonium were originally found during the cooler times of the year in inland Polish waters (Figure 2.4). Other species (e.g., Cladophora, Oedogonium, Ulva) appear to be restricted to the warmer periods of the year, like summer or beginning of autumn. In contrast, representatives of Microspora have a wide tolerance to temperature and appear in the same locations from spring to summer and gradually disappear with the beginning of autumn

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Apr May Jan Jul Aug Sep Oct

Apr May Jan Jul Aug Sep Oct

Tribonema

Cladophora

3

3

2

2

1

1

0

0 Oedogonium

Ulothrix 3

3

2

2

1

1

0

0

Ulva

Vaucheria 3

3

2

2

1

1

0

0 Microspora

Rhizoclonium 3

3

2

2

1

1

0

0

Figure 2.4 Relative seasonal abundance of filamentous green algae in inland ecosystems from Poland (0 – absent; 1 – present; 2 – common; and 3 – abundant).

(Figure 2.4). In reservoirs in which filamentous algae occurred, the highest population was recorded a neutral pH and under slow water movement [33, 34, 96]. One should also pay attention to the replacement of species within the growing season, which could be interpreted as a result of interspecific competition. The composition of filamentous green algae shows clear similarities in freshwater ecosystems in Poland and other parts of Europe, such as Great Brittan [97], or southern Spain [98]. Although some species may have a very wide, cosmopolitan distribution, not all places can create such a massive material in the form of visible mats. Ecosystems in which macroscopic algae form distinct mats are often characterized by high levels of chlorides and nutrients such as nitrogen or phosphorus [99, 100]. In particular, representatives of Ulva genus inhabit reservoirs that have

2.5

Summary

a constant supply of anthropogenic pollution such as chlorides that originate during road cleanings (NaCl) during winter and flow with the rain water to roadside watercourses. The appearance of filamentous algae may also be associated with the presence of heavy metals such as Cd, Cu, Ni, Pb, Zn, and Mn. There monitoring is helpful in evaluating the state of environment and pollution [101, 102]. According to research, the accumulation of at least one of these metals, for example, Cu, by Ulva is dependent on the salinity of the water body. Reduced salinity increases the toxicity of Cu on the thallus of macroscopic algae. Conversely, when the water salinity is higher, a less negative metal impact on this macroalgae is observed [103]. Obtained data reveal that the average preferred concentration of NaCl for U. compressa L. amounts to 378 mg l−1 . This amount is far more than for U. flexuosa Wulfen (77 mg l−1 ) and U. intestinalis L. (86 mg l−1 ). Similar differences occurred in the average concentrations of NO3 − ions (U. intestinalis and U. flexuosa prefer higher levels) and PO4 − ions (U. intestinalis, U. compressa). The lowest variations of preferences for each species were found in the values of the average water pH: U. compressa (8.1), U. intestinalis (7.6), and U. flexuosa (7.5) [94–96, 99]. These ecological results clearly indicate that freshwater Ulva cultures should resemble as much as possible the habitat requirements of given species for a determined composition of the medium. Individual modifications might be needed. The extensive blooms of macroalgae in the form of mats floating freely on the water surface affect the biochemical and physical properties of the aquatic ecosystem. In consequence, it also affects other organisms that inhabit them [104]. This mainly concerns the relationship between temperature and the presence of the mat, access to light and oxygen (which is more difficult), as well as the pH and concentration of nutrients [105, 106]. The rapid growth of filamentous green algae and their subsequent decay and fall to the bottom may be contributing to the unfavorable conditions for the development of water macrofauna and phytoplankton [105]. This excessive amount of biomass in some cases can be effectively reduced by herbivorous organisms [106], but it is difficult to get such an effect with a large and dense algal patches.

2.5 Summary

There is a need to synthesize the data available for understanding the autecology of particular macroalgal species in inland waters and their possibilities to produce a large biomass under natural conditions, rather than proposing more and more elaborate scenarios to explain evolutionary aspects. Thus, the systematic occurrence of macroalgae blooms in the form of mats in freshwater raises the possibility of using the readily available biomass as a raw material in various sectors of the economy and industry. For industrial purposes, products enriched with extracts derived mostly from marine macroscopic algae are of importance[107]. However, more and more research is conducted in the direction of using extracts from the thalli of the freshwater macroalgae for practical purposes. Algae materials used

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in cosmetics or food industry should be of high quality and use efficient tools. This creates great opportunities toward improving the culture of select species on a commercial scale. Thus, general data on the biology and ecology of freshwater macroscopic algae will contribute to the development of the best methods to make this group of algae useful on the industrial scale.

Acknowledgments

This project is financed in the framework of grant entitled – Innovative technology of seaweed extracts – components of fertilizers, feed, and cosmetics (PBS/1/A1/2/2012) attributed by The National Centre for Research and Development in Poland. References 1. Guiry, M.D. (2012) How many species

2.

3.

4.

5.

6.

7.

8.

of algae are there? J. Phycol., 48, 1057–1063. De Clerck, O., Bogaert, K.A., and Leliaert, F. (2012) Diversity and evolution of algae. Adv. Bot. Res., 64, 55–86. Van den Hoek, C., Mann, D.G., and Jahns, H.M. (1995) Algae. An Introduction to Phycology, 1st edn, Cambridge University Press, 623 pp. Chapman, R.L. (2013) Algae: the world’s most important “plants”-an introduction. Mitig. Adapt. Strateg. Global Change, 18, 5–12. Mareš, J. (2009) Combined morphological and molecular approach to the assessment of Ulva (Chlorophyta; Ulvophyceae) in the Czech Republic. MSc Thesis. Faculty of Science, University of South Bohemia, Czech Republic, 72 pp. Rajkuniar, R. and Yaakob, Z. (2013) in Biotechnological Applications of Microalgae. Biodiesel and Value-Added Products (ed. F. Bux), pp. 7–16. Guiry, M.D. and Guiry, G.M. (2013) AlgaeBase, World-wide Electronic Publication and National University of Ireland, Galway, http://www.algaebase.org (accessed 15 November 2013). South, R.G. and Whittick, A. (1996) Introduction to Phycology, Blackwell

9.

10.

11.

12.

13.

14.

15.

16.

Scientific Publications, Oxford, London, Edinburgh, 341 pp. Bellinger, E.G. and Sigee, D.C. (2010) Freshwater Algae. Identification and Use as Bioindicators, 1st edn, WileyBlackwell, Ltd, Chichester, 271 pp. Starmach, K. (1972) in Flora słodkowodna Polski (ed. K. Starmach), Państwowe Wydawnictwo Naukowe, Warszawa, Kraków, p. 163. Pliński, M. and Hindák, F. (2012) Zielenice – Chlorophyta: Filamentous Green Algae, Wydawnictwo Uniwersytetu Gdańskiego, Gdańsk, 189 pp. Messyasz, B. and Pikosz, M. (2014) Massive blooms of metaphyton in small ponds in the Wielkopolska Region. Acta Agrobot., Dissertation. 68 (4), 82–91 Andersen, R.A. (2005) Algal Culturing Techniques, Elsevier Academic Press, London, pp. 31–21, 425–500. Hiraoka, M. and Oka, N. (2007) Tank cultivation of Ulva prolifera in deep seawater using a new “germling cluster” method. J. Appl. Phycol., 20, 97–102. Robertson-Andersson, D., Potgieter, M., Hansen, J., Bolton, J.J., Troell, M., Anderson, R., Hallin, C., and Probyn, T. (2008) Integrated seaweed cultivation on an abalone farm in South Africa. J. Appl. Phycol., 20, 579–595. Cruz-Suarez, L.E., Leon, A., Pena-Rodriguez, A., Rodriguez-Pena,