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The University of New South Wales Faculty of Applied Science School of Biological Technologies Department of Food Science and Technology THE UNIVERSITY OF NEW SOUTH WALES FACULTY OF APPLIED SCIENCE SCHOOL OF BIOLOGICAL TECHNOLOGIES DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY UTILISATION OF THE AUSTRALIAN JELLYFISH CATOSTYLUS SP. AS A FOOD PRODUCT Presented as a Thesis for the degree of Doctor of Philosophy by Lucita G. Suelo B.Sc.(VISCA), M.Sc.(UPLB), M.App.Sc. Submitted Sydney, October 1986 The candidate, Lucita G. Suelo hereby declares that none of the work presented in this thesis has been submitted to any other University or Institution for a higher degree. i ACKNOWLEDGEMENT I wish to express my heartfelt thanks and profound gratitude to the following persons/entities who have extended their support while I was doing my postgraduate study: Dr Michael Wootton, my supervisor, for his guidance, unlimited help and valuable suggestions during the experimental and preparation of this thesis; The Australian Government through the Australian Development Assistance Bureau (ADAB) for granting the scholarship under the Colombo Plan and the University of New South Wales which made this study possible; The Philippine Government through the Panay State Polytechnic College for granting me a study leave to pursue a postgraduate degree; The UNSW, Department of Food Science and Technology staff headed by Prof. R.A. Edwards for their assistance during the duration of my stay in the University; The School of Zoology through Dr Macintyre for allowing the use of the boat for fishing; Prof. McGilchrist of the School of Mathematics for his assistance in the statistical analysis; Dr Penny Farrant of the School of Botany for her help in the light microscopy and Sergei Kouprach of the SEM unit for his assistance in freeze-fracture and photoelectron microscopy; Mrs Rose Varga of the Dept. of Biotechnology for her tutorship in microtechniques and use of microtome; The Fishing Industry Research Committee for partial financial support of this research; ii Ms Cherelle Czerney for typing this thesis; My fellow postgraduate students.at the Department of Food Science and Technology especially Phaisan for his help in graphing, Lubis in statistical analysis, Pairat, Made, Hari, Redi, Kartini, Sutardi, Haryadi, Noryati, Elli and many others who made my stay a pleasant experience; My parents, sisters and brothers for their love, understanding and prayers while I was undertaking this study; and Libor, for his patience, unlimited help, love and understanding, this work is humbly dedicated. iii TABLE OF CONTENTS Page no Acknowledgement i Table of Contents iii Abstract ix 1. Introduction 1 2. Literature Review 6 2.1 Zoological Classification of Jellyfish 6 2.1.1 Phylum Cnidaria 6 2.1.2 Taxonomy 8 2.1.3 Semaeostomae and Rhizostomae species found in Australian waters 10 2.1.4 Life cycle 11 2.1.5 Regeneration 13 2.1.6 Ecology/Adaptation 13 2.1. 7 Availability/Distribution 14 2.1.8 Seasonal distribution of jellyfish in Australian waters 18 2.2 Studies on the Processing of Jellyfish 23 2.3 Utilisation as Food 24 2.4 Chemical Properties of Unprocessed Jellyfish 26 2.4.1 Moisture content 26 2.4.2 Protein and amino acids 27 2.4.3 Lipid 34 2.5 Preservative Mechanism of the Traditional Jellyfish Processes 38 2.6 Disposal of Brine for Jellyfish Processing 39 3. Materials and Methods 40 3.1 Materials 40 3.1.1 Jellyfish samples 40 3.2 Methods 41 3.2.1 Processing of jellyfish 41 3.2.1.1 Traditional method 41 3.2.1.2 Shortened method 42 iv Page 3.2.1.2.1 Brining and drying technique 42 3.2.1.2.2 Blanching and drying technique 42 3.2.2 Analysis of the yield, chemical and organoleptic properties of processed jellyfish 42 3.2.2.1 Yield 42 3.2.2.2 Chemical properties 43 3.2.2.2.1 Moisture content 43 3.2.2.2.2 Water activity 43 3.2.2.2.3 Protein 44 3.2.2.2.4 Fat (acid hydrolysis) 45 3.2.2.2.5 Ash (dry ashing) 46 3.2.2.2.6 pH 46 3.2.2.2.7 Sodium chloride (Volhard method) 46 3.2.2.2.8 Aluminium 48 3.2.2.2.9 Total volatile acidity 49 3.2.2.2.10 Titratable acidity 49 3.2.2.2.11 Total volatile bases (TVB) 49 3.2.2.2.12 Trimethylamine (TMA) 50 3.2.2.2.13 Trimethylamine oxide (TMAO) 50 3.2.2.2.14 TCA soluble nitrogen 51 3.2.3 Organoleptic properties 51 3.2.3.1 Panel evaluation of texture and flavour 52 3.2.3.2 Flavour, texture and overall acceptability evaluation 52 3.2.4 Solubility studies 53 3.2.4.1 Solubility on O.lM potassium chloride 53 3.2.4.2 Solubility on heating in sodium dodecylsulphate + B-mercaptoethanol 53 3.2.4.3 Solubility of protein in sodium chloride solution 53 3.2.4.4 Solubility of protein using enzymes 54 3.2.4.4.1 Pepsin solubility 54 3.2.4.4.2 Trypsin solubility 54 3.2.4.4.3 Pepsin-trypsin solubility 54 3.2.4.5 Solubility of sample in water 54 V Page 3.2.5 Extractability studies 55 3.2.5.1 Effect of sodium chloride on extractability of protein in jellyfish 55 3.2.5.2 Effect of temperature on extractability of protein in jellyfish 55 3.2.5.3 Effect of pH on protein extraction efficiency 56 3.2.6 Digestibility of jellyfish protein by pepsin 56 3.2.7 Isoelectric focusing 57 3.2.7.1 Materials 57 3.2.7.2 Reagents 58 3.2.7.3 Sample preparation 58 3.2.7.4 Methods 59 3.2.7.4.1 Casting the gel 59 3.2.7.4.2 Moulding 60 3.2.7.4.3 Running the gels 60 3.2.7.4.4 Fixing and staining 61 3.2.7.4.5 Determination of isoelectric point (pi) 61 3.2.7.4.6 Densitometric analysis 62 3.2.8 Amino acid analysis in jellyfish 63 3.2.8.1 Acid hydrolysis 63 3.2.8.2 Sample preparation for chromatography 63 3.2.8.3 Reagent preparation 64 3.2.8.3.1 Ninhydrin 64 3.2.8.3.2 Buffer A (trisodium citrate dehydrate) 65 3.2.8.3.3 Buffer B (trisodium citrate dehydrate) 65 3.2.9 Structural examination of jellyfish tissues 66 3.2.9.1 Light microscopy 66 3.2.9.2 Freeze-fracture technique and electron photomicroscopy 66 3.2.10 Brine microbiological assay 67 vi Page 3.2.11 Brine recycling 67 3.2.12 Microbial changes during spoilage of fresh jellyfish 68 4. Results and Discussion 69 4.1 Chemical Composition and Properties of Fresh Jellyfish 69 4.1.1 Proximate analysis 69 4.1.2 Total amino acids 71 4.1.3 Isoelectric focusing of proteins 74 4.1.4 Soluble solids, solubility and digestibility of proteins 76 4.2 Preliminary Study of the Traditional Processing Method 76 4.2.1 Effect of alum and salt as preservatives 76 4.2.1.1 Alum alone 78 4.2.1.2 Salt alone 78 4.2.1.3 Alum-salt combination 79 4.2.2 pH of jellyfish and preservatives used during processing 84 4.3 Optimisation of Salt Levels 85 4.3.1 Moisture content, a and weight loss w 86 4.3.2 Protein 87 4.3.3 pH 88 4.3.4 Salt and aluminium levels 88 4.3.5 Organoleptic properties 91 4.3.5.1 Texture 92 4.3.5.2 Flavour 92 4.3.6 Protein in brine 94 4.4 Effect of Processing Time on the Properties and Yield of Jellyfish 96 4.4.1 Changes during the first brining step 97 4.4.2 Changes during the second brining step 97 4.4.3 Drying time 100 4.4.4 Organoleptic evaluation of jellyfish 100 vii Page 4.5 Approaches for Rapid Processing of Jellyfish 102 4.5.1 Dry salting of jellyfish 103 4.5.2 Artificial drying 103 4.5.3 Brine properties and recycling 104 4.5.4 Artificial drying of unsalted jellyfish 108 4.5.5 Recommended rapid process for jellyfish preservation 109 4.5.6 In vitro digestibility of jellyfish proteins 111 4.6 Solubility Properties of Processed Jellyfish Proteins 112 4.6.1 Protein extractability 113 4.6.1.1 Influence of pH 113 4.6.1.2 Influence of salt concentration 115 4.6.1.3 Influence of temperature 117 4.6.2 Solubilisation by O.lM KCl 119 4.6.3 Solubilisation by SDS + B-mercapto­ ethanol 119 4.6.4 Solubilisation by pepsin 120 4.6.5 Amino acid composition of processed jellyfish 122 4.6.6 Isoelectric focusing analysis of proteins from processed jellyfish 126 4.6.1.1 Isoelectric focusing analysis of protein from brine 132 4.7 Physical Structure of Jellyfish 135 4.7.1 Fresh jellyfish 136 4.7.1.1 Freeze~fracture and electron microscopy 136 4.7.1.2 Light microscopy of fresh jellyfish 137 4.7.2 Effects of processing on jellyfish tissues 146 4.7.2.1 Effects of alum alone and drying 146 4.7.2.2 Effects of salt alone and drying 149 4.7.2.3 Effects of salt-alum combination and drying 149 4.8 Chemical and Microbial Changes During Storage and Processing of Fresh Jellyfish 152 viii Page 4.8.1 Fresh jellyfish 155 4.8.1.1 Trimethylamine oxide, trimethylamine, total volatile base and pH 155 4.8.1.2 Total plate count 157 4.8.1.3 TCA soluble nitrogen 159 4.8.2 Changes in TMA and TVB during the rapid processing of jellyfish 160 4.8.3 Total volatile acidity and titratable acidity of processed jellyfish 160 4.8.3.1 Rapid process jellyfish 161 4.8.3.2 Traditional process jellyfish 163 5.
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