ATLANTIC FISHERIES TECHNOLOGY CONFERENCE

ATLANTIC FISHERIES TECHNOLOGY CONFERENCE

TABLE OF CONTENTS

PROGRAM AT A GLANCE .............................................................................................1 PRE-CONFERENCE .........................................................................................................5 Abstracts ...................................................................................................................9 SCIENTIFIC PROGRAM .................................................................................................19 Abstracts ...................................................................................................................25 POSTERS ............................................................................................................................47 Abstracts ...................................................................................................................49

55th Annual Atlantic Fisheries Technology Conference 1











PRE-CONFERENCE ABSTRACTS

September 7, 2010 (1) RAW MATERIAL SOURCES FOR THE LC OMEGA 3 FATTY ACID MARKET,

TRENDS AND SUSTAINABILITY Anthony P. Bimbo (USA) International Fisheries Technology, 55 Cedar Lane, Post Office Box 1606, Kilmarnock, Virginia, USA 22482

On an almost daily basis now, some article appears in the popular or scientific press declaring a growing shortage of fish oil in its key global markets. Reports and forecasts of El Nino events, earthquakes in Peru, and Chile, volcanic eruptions in Iceland and environmental predictions that ocean fish are being pursued to extinction add to the doom and gloom situation. While most of the concerns have been raised in regard to the aquaculture industry where fish oil and fishmeal are highly desired products, questions have also been raised from groups within the nutritional oils and nutraceutical value chain about the sustainability of the fish oil resources. While the questions related to aquaculture are routinely addressed, the issue of raw materials for the long chain (LC) omega 3 nutritional oils or nutraceutical market have only been addressed briefly. This presentation will attempt to put the raw material sources for the omega 3 fish oil value chain into perspective. (2) MARINE OIL PRODUCTS AND FOOD APPLICATIONS Robert Orr (Canada)

Chairman and Co-Founder, Ocean Nutrition Canada Ltd., 101 Research Drive Dartmouth, Nova Scotia, B2Y 4T6 Canada

Marine oils, primarily in the form of fish oils and algal oils containing Omega3 Fatty Acids (EPA &DHA) have now become an important ingredient in the dietary supplement and functional (healthy) food sectors. Global annual sales of marine oil ingredients have reached the $1 billion level and the retail sales of products containing marine oils is now more than $8 billion annually. The food ingredient sector and regulators are just now beginning to understand the importance of Omega 3s in the diet and the developing strategies to impact the significant dietary deficiency of Omega 3s that exist in the western diet. New delivery technologies such as micro-encapsulation are required to optimize grow potential in the food sector. With these food technology enhancements, continued regulatory intervention and a new learning on the part of multinational food companies in the area of marketing “Healthier Products”, we can forecast that marine oils containing Omega 3s will continue to grow at a double digit rate for at least the next 5 years.


(3) CHEMISTRY AND HEALTH BENEFITS OF MARINE OILS AND THEIR NOVEL OMEGA-3 DERIVATIVES Ying Zhong, Freidoon Shahidi (Canada) Departments of Biochemistry and Biology and Ocean Sciences Centre, Memorial University of Newfoundland, St. John’s, NL, Canada A1B 3X9

Long-chain polyunsaturated fatty acids (PUFA) originating from marine sources, including marine algae, fish and mammals, have been recognized for their health benefits to humans. The omega-3 fatty acids eicosapentaenoic (EPA), docosapentaenoic (DPA) and docosahexaenoic (DHA) acids in marine oils or their concentrates have been proven to play an important role in the prevention and treatment of cardiovascular disease, inflammatory and autoimmune disorders, cancer and neurodegenerative diseases. More recently, these fatty acids have been studied for novel applications in food, cosmetic and nutraceutical industries in combination with plant-derived polyphenols. Long-chain PUFA were incorporated into a tea polyphenol epigallocatechin gallate (EGCG) in order to improve the lipophilicity of the water-soluble EGCG and thereby enhancing its bioavailability and expanding its application in lipophilic media. The EGCG-PUFA esters exhibited excellent antioxidant, anti-inflammatory, antiviral and anticancer activities, some of which were greater than that of the EGCG or PUFA alone. Combination of polyphenols and PUFA into one molecule provides a good approach to designing novel nutraceuticals for health promotion and disease risk reduction. (4) PROCESSING OF MARINE OILS AND TOTAL REMOVAL OF

ENVIRONMENTAL CONTAMINANTS R. Verhé1, J. Vila Ayala2, W. De Greyt2 (Belgium) 1Ghent University, Belgium; 2De Smet Ballestra, Zaventem, Belgium Interest in the functionality and health beneficial properties of marine oils continue to grow each year. In order to use marine oils as food products or animal feed, the oils have to undergo processing and refining. This involves various steps including degumming to remove phospholipids and other gums. Bleaching, using various adsorbents, eliminates coloring substances and minerals which are acting as pro-oxidants. Free fatty acids can be discarded by neutralization or stripping. The last step in refining is a deodorization process at high temperature in vacuum and using steam for the elimination of oxidation products and the remaining free fatty acids. Marine oils from certain regions are contaminated by dioxins, benzofurans and polychlorinated biphenyls. These contaminants are eliminated for 95% by a combined process involving an adsorption step using active carbon and a stripping process at 190 °C. Using this procedure, the concentration of the polyunsaturated fatty acids is slightly decreased and produces oils suitable for human consumption or animal feed.


(5) ALGAL OILS: A PROMISING SOURCE OF OMEGA-3 FATTY ACIDS Roberto E. Armenta (Canada) Ocean Nutrition Canada, Ltd., Dartmouth, Nova Scotia, Canada

Omega-3 fatty acids, such as docosahexaenoic acid and eicosapentaenoic acid, can be produced by some algae strains. Lately, research on microbial sources for production of omega-3 fatty acids has increased significantly. Besides producing algal oils in a controlled environment, an advantage of algal oils is that they contain antioxidant components that may protect omega-3 fatty acids against oxidation. Commercial viability for producing omega-3 algal oils is determined by microbial oil production yields which are defined by effective upstream (fermentation) and downstream (oil extraction) processes. This paper presents advances in the production of algal omega-3 rich oils. (6) PROTEIN HYDROLYSATES FROM MARINE RESOURCES

Dr. Hordur G. Kristinsson (Iceland) Acting CEO, Director, Biotechnology and Biomolecule Division, MATIS - Icelandic Food and Biotech R&D, Reykjavik, Iceland

Every year significant amounts of aquatic food raw materials go underutlized. These raw materials are a rich source of high quality proteins, which with proper extraction techniques and processing can find use in foods, specialty feeds, neutraceuticals, cosmeceuticals and medical products. The market for these products is growing and their production leads to a very significant value addition of the raw materials used. Novel processes and techniques have been developed by the author and his collaborators where highly purified functional proteins are extracted from various byproducts and further processed into peptides of various molecular weights. These peptides have demonstrated high bioactivity in various in-vitro models. This includes the ability to inhibit the angiotensin I converting enzyme, strong free radical scavenging ability as well as good ability to chelate metals and high reducing power. In addition, the peptides have been shown to inhibit lipid oxidation in food systems, thus showing good potential as natural food antioxidants. Human and animal clinical trials are also ongoing with select peptide products. Currently the bioactive peptides are being moved from pilot to commercial stage and represents a promising way to utilized previously poorly or unutilized raw materials.


(7) SEAWEED AND ITS BIOACTIVE COMPOUNDS Kazuo Miyashita (Japan) Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611, Japan

Lipid related compounds produced by marine organisms are unique and different from those obtained from terrestrial animals or plants. Omega-3 polyunsaturated fatty acids (PUFA) such as EPA (20:5n-3), DPA (22:5n-3), and DHA (22:6n-3) are well-known marine nutraceutical lipids. Much attention has also been paid to novel biological activities of marine carotenoids such as astaxanthin and fucoxanthin. Brown seaweed lipids are good source for both fucoxnathin and omega-3 PUFA such as 18:4n-3 (stearidonic acid) and EPA. This presentation will give an overview on the nutraceutical functionality of brown seaweed lipids and their effective utilization. (8) SEAL BLUBBER OIL AND SEAL-BASED PRODUCTS Fereidoon Shahidi, Jiankang Wang (Canada)

Departments of Biochemistry and Biology, Memorial University of Newfoundland, St. John's, NL, Canada A1B 3X9

Seal blubber oil is an important marine oil that contains omega-3 fatty acids mainly eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). The omega-3 fatty acids are located primarily in sn-1/3 positions of triacylglycerols while EPA and DHA are located primarily in sn-2 position of fish oils; DPA is much less prevalent in fish oils. In addition, seal blubber oils contain much less C14:0 than fish oils which may be considered advantageous. Thus, seal blubber oil is quite important in health promotion and disease risk reduction and DPA is instrumental in preventing the fragility of red blood cells. In terms of stability characteristics, seal blubber oil is more stable than fish oils and hence can be easily employed in food product development with extended shelf-life.


(9) PROCESSING OF MARINE AND ALGAL OILS AND PRODUCTION OF EPA, DPA AND DHA CONCENTRATES Udaya N. Wanasundara (Canada) Senior Scientist and Project Leader, POS Pilot Plant Corporation, 118 Veterinary Road, Saskatoon, SK, S7N 2R4, Canada

Crude oil from marine and algal sources contains considerable amounts of non-triacylglycerol (non-TAG) substances. Some of these compounds are beneficial (i.e. tocopherols as an antioxidant), however, majority (i.e. free fatty acid, colored substances, phospholipids, glycolipids, etc) are objectionable because they render off color and off odor in the oil and cause foam and precipitates during storage. Presence of non-TAGs decreases storage stability and shelf life of the oil. Therefore, the processing of crude marine oils comprising a series of discrete steps such as degumming, alkali refining, bleaching and deodorization (RBD) is carried out to remove these impurities. The most prevalent polyunsaturated fatty acids of marine oils are docosahexaenoic (DHA), docosapentaenoic (DPA) and eicosapentaenoic (EPA) which are being used in products ranging from dietary supplements to infant formulas. Naturally, these fatty acids are associated with other lipophilic compounds of the oil and effective separation and isolation techniques are needed to recover them in concentrated forms to use in nutritional applications. At POS Pilot Plant Corp. (Canada), we are capable to process highly unsaturated crude oil into stable RBD oil and also to produce concentrated forms of EPA and DHA in small- or large-scale quantities via different techniques. This presentation focuses on our process capabilities and new methodologies that we’ll be able to provide. (10) MARINE AND OTHER OILS IN SPECIALTY APPLICATIONS IN BIOFUEL

PRODUCTION R. Verhé1, V. Van Hoed1, C. Echim1,2, W. De Greyt2 (Belgium)

1Ghent University, Belgium; 2De Smet Ballestra, Zaventem, Belgium The conversion of oils and fats for the production of biodiesel is a major initiative for the substitution of petroleum derived fuels. The majority of the lipid feedstocks are edible oils which is creating a confrontation food versus fuel. In addition, the use of edible lipids has given rise to higher food prices. Therefore, alternative feedstocks and/or technologies have been developed leading to biodiesel of the second generation. Alternative feedstocks are non-edible oils, algae oil, waste animal fats (e.g. marine oils), used frying and cooking oils and side-streams from refining. As the production cost of biodiesel is for 85% dependent on the feedstock, search for cheaper lipid resources is crucial. However, the use of these alternative resources involves additional processing or refining steps, such as combined transesterification/esterification, more severe refining and post- purification, in order to meet biodiesel standards. Several alternative feedstocks, including marine oils, for the production of biodiesel will be illustrated using a variety of processing techniques. A comparison of biodiesel produced from crude fish oils with other crude or waste oils will be presented. Crude animal fats and marine oils not suitable for food and feed can be used for the production of heat and power (CHP) using stationary diesel engines. This new application will illustrated by a power plant of 12.5 MW in Belgium. The various purification steps of the feedstocks will be discussed. Up to now, marine oils are not widely used as a feedstock for biofuels, but oils not suitable for consumption are looking promising for the generation of green energy.


(11) MARINE NATURAL PRODUCTS AS PHARMACEUTICALS Russell Kerr (Canada)

Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue Charlottetown, PE, Canada C1A 4P3

Medicinal agents derived from nature have been utilized for thousands of years to treat a myriad of ailments ranging from general aches to infections. The use of such natural products is documented as early as the ancient Egyptians, yet despite the antiquity of their use, it wasn’t until the 19th century that scientists began identifying the active pharmaceutical ingredients of botanical extracts. In 1928 Fleming discovered the first natural product of microbial origin, and the ensuing industrialization of penicillin during World War II represented a seminal step in the use of a single natural product as a pharmaceutical entity. The field of marine natural products has evolved out of ground-breaking work in the 1970s and now represents a significant source of pharmaceutical leads. Two marine natural products have been approved for human use and numerous others are in human clinical trials. (12) COLLAGEN AND GELATIN FROM SEAFOOD PROCESSING BY-PRODUCTS Joe Regenstein1 (USA), Reyes Pla2 (Spain)

1Cornell University, Department of Food Science, Ithaca, New York 2Tecnologia dels aliments, Departament de ciència animal i dels aliments, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain

Gelatin is a class of protein fractions that has no existence in nature, but is derived from the parent protein collagen, by procedures involving the destruction of crosslinkages between polypeptide chains of collagen and breakage of polypeptide bonds. Most commercial collagens and gelatins are obtained from bovine bone, bovine hide, and pork skin. In recent years collagen and gelatin obtained from fish processing byproducts have become of interest, in part due to the requirements for a kosher and halal ingredient and in part due to consumer concerns about bovine spongiform encephalopathy (BSE, “mad cow disease”) as well as being a way to increase the economic return to the fishing industry. About 30% of the waste from fish processing after filleting consists of skin, bone, and scales with high collagen content that could be used to produce collagen and gelatin. The yield and properties of collagen and gelatin are influenced by the source of the raw material, the nature of and concentration of the acid or alkali used during pretreatment, the temperature and time of pretreatment and during extraction, and other process variables. These in turn, determine the properties, such as gel strength and viscosity, of the different fish gelatins.


(13) QUALITY PRESERVATION OF MARINE OILS AND SEAFOODS Charlotte Jacobsen1, Caroline P. Baron (Denmark)

Lipid and Oxidation Group, National Food Institute, Technical University of Denmark, Søltofts Plads, Bygn. 221, 2800 Kgs. Lyngby 1Corresponding author: [email protected]

There is an increasing interest in developing healthy ingredients, functional foods as well as high quality fish products. Indeed a high intake of polyunsaturated lipids such as omega-3 fatty acids have been shown to be important in order to keep a healthy and balanced diet. However, foods containing omega-3 fatty acids are very susceptible to lipid oxidation, which will give rise to undesirable off-flavours and unhealthy reaction compounds. Hence, efficient strategies to prevent lipid oxidation are required. Unfortunately, lipid oxidation and antioxidant mechanisms in food matrices are very complex and many factors can influence the rate of lipid oxidation and the efficacy of different antioxidants in food matrices. This includes for example the following factors: the pH, the presence of metal ions (iron, copper, etc…), and type of emulsifier. Because of this complexity it is difficult to predict the behaviour and efficacy of antioxidants in complex food systems such as fish products, and omega-3 enriched food emulsions such as milk, salad dressings, and mayonnaise. This presentation will discuss different strategies to prevent lipid oxidation in fish products as well as in omega-3 enriched foods including optimisation of processing conditions, and choice of ingredients (oil, emulsifier and antioxidants). As for the addition of antioxidants the efficacy of different antioxidant principles and mechanisms will be summarised from our own data sets. This includes for example discussion of the efficacy of different antioxidants such as caffeic acid, ascorbyl palmitate, tocopherol, propyl gallate and EDTA in emulsions and in fish products. (14) CHITIN, CHITOSAN, CHITOOLIGOSACCHARIDES AND THEIR

DERIVATIVES Se-Kwon Kim (Republic of Korea)

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

Chitin and chitosan have been of interest during past few decades due to their biological activities and various industrial applications. However, they have several drawbacks to be utilized in biological applications, including poor solubility under physiological conditions. COS, partially hydrolyzed products of chitosan are of great interest in pharmaceutical and medicinal applications due to high solubility and nontoxicity. Moreover, their derivatives have opened new avenues for its applications. COS and their derivatives have been reported to have various biological activities, such as antioxidant, anticancer, anti-inflammatory, anti-obesity, antimicrobial, anti-HIV and antihypertensive effects and these various biological activities are due to their physico-chemical properties. Many interests and efforts are still ongoing for development and application of chitin, chitosan, COS and their derivatives. In this section, we have summarized biological activities and considerable potential in useful applications of COS and their derivatives referring to our previous studies.








Atlantic Fisheries Technology Conference



ORAL ABSTRACTS

September 8, 2010 (1) MANAGEMENT, CONSERVATION AND ECOLOGICAL BALANCE IN THE

HARVEST OF FISH STOCKS George A. Rose (Canada)

Fisheries and Marine Institute of Memorial University, St. John’s, NL, Canada A1C 5R3 Wild fisheries worldwide are under increasing pressure to produce more, but productive capacity is unlikely to exceed 100 million tonnes. Many stocks are currently overfished or below maximum productivity, and changing with climate. Conservation interests demand that non-commercial species not be disproportionately impacted by fisheries, with some crucial to harvested species productivity. Consumers demand product be from sustainable fisheries and correctly identified to species and stock, yet public concerns can be biased, favouring some species over others. As a result of these competing demands, management goals are often unclear, but for fisheries the key question is how to achieve maximum sustainable productivity by balancing various fisheries. For conservation, the key question is how to maintain biodiversity in a balanced ecosystem. It is not clear if these questions have the same answer. A more diverse and experimental approach is needed – only then will goals for marine ecosystems be clearer.

(2) MARINE BIOPROSPECTING FOR NATURAL PRODUCTS – THE

CHALLENGES AND OPPORTUNITIES Russell Kerr (Canada) Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue,

Charlottetown, PEI, Canada C1A 4P9

The field of marine natural products has proven to have great applications in the development of new pharmaceutical agents. Two such compounds have recently been approved for use in humans – one is a powerful cancer drug and the other is used as a morphine replacement. Many other marine natural products are in human clinical trials for varied applications and it is anticipated that additional drugs will be approved in the coming years. One key challenge hampering the development of many marine natural products is the limited amounts of natural product that are often available. Potential solutions to this “supply issue” will be discussed as well as a new strategy to discover marine natural products that can be produced in a sustainable manner.


(3) CHALLENGES IN SUPPLY, COMMERCIALIZATION AND MARKETING OF MARINE NUTRACEUTICALS

Robert Orr (Canada) Chairman and Co-Founder, Ocean Nutrition Canada Ltd., 101 Research Drive, Dartmouth, Nova Scotia, B2Y 4T6 Canada

For the last decade much of the focus in nutra-science, natural health product and nutraceuticals sectors has been on “Innovation”. However this “Innovation” focus has lead primarily to investments in R&D and most of that in the area of new compound discovery. However when it comes to building Sustainable Competitive Advantage for products or companies there is a need to focus on more than just Innovation. To create financial success in the marine, natural products, sector requires organizations to build competencies beyond innovative research and new compound discovery. Financial success also requires skills in Delivery Technologies, Resource and Supply Chain Management, Regulatory Affairs, Marketing, and Commercialization. In addition, each of these competencies must be developed and implemented in a global context. Our greatest challenge is not the discovery and development of new ideas and products it is the capacity to “get them to customers” and commercialize them in a globally competitive marketplace. Utilizing the experience gained from the building of Ocean Nutrition Canada Ltd., from an idea on a piece of paper into a global leader in the Omega 3 ingredients sector, Robert Orr will explain how to effectively apply these six competencies to meet the commercialization challenges of today’s marketplace. In addition to the six competencies there are two other key components that are the catalysts for commercial success, in this area. Those components are leadership and a supportive, enabling environment or support community. The question for all of us is not “can we learn about these 6 competencies and 2 catalyzing components?” but can we build the systems that can replicate past success and make future success more predictable. (4) BIOTECHNOLOGY AND GENOMICS IN FISHERIES AND AQUACULTURE

RESEARCH Matthew L. Rise (Canada)

Assistant Professor and Canada Research Chair in Marine Biotechnology, Ocean Sciences Centre, Memorial University of Newfoundland, St. John’s, NL, A1C 5S7

With my collaborators and trainees, I build and use functional genomics tools [e.g. suppression subtractive hybridization (SSH) cDNA libraries for targeted gene discovery, and DNA microarrays for global gene expression profiling] to study the genes and molecular pathways involved in fish responses to pathogens (e.g. bacterial and viral), enhanced growth, and environmental stressors (e.g. temperature stress and pollutants). In addition to providing valuable new information on the genetic basis of these biological processes, this research will likely lead to new strategies for improving the health of farmed and wild fish.


(5) FISHMEAL AND FISH OIL UPDATE 2010: TURMOIL AND TRANSITION SINCE 1998 Anthony P. Bimbo (USA) International Fisheries Technology, 55 Cedar Lane, Post Office Box 1606, Kilmarnock, Virginia, USA 22482

At the 43rd AFTC held in St. Johns, Newfoundland we faced turmoil amid transitions in the fishmeal and fish oil industry. Fish caught specifically for fishmeal and oil production accounted for about 25-30% of the world’s landings. Since the aquaculture industry was growing quite rapidly there were concerns about the supply of fishmeal and oil and could they keep pace with the growth in aquaculture. 1997 and 1998 were also plagued with a major El Nino weather event which played havoc with the South American fisheries and that in turn impacted the supply and demand for fishmeal and oil and their prices on the world market. While having prices at record highs (actually unprecedented) appeared to be an advantage, it was actually a 2 edged sword. Pressures from environmental groups concerned with resource sustainability, health issues concerned with trans fatty acids in partially hydrogenated fats and the potential health benefits of the omega 3 fatty acids in fish oil all conspired to put the fishmeal and oil industry into a state of transition. The high prices caused dramatic changes in fishmeal consumption patterns and fostered research to find less expensive replacements. So what has happened since the 1998 AFTC Conference? In 2009 we had a relatively minor El Nino event, environmental groups and various NGO’s continue to raise concerns about the viability of the fish resources. Fishmeal and oil only account for about 15-18% of the world’s fish catch, yet aquaculture continues to grow and must grow to keep up with demand for edible fish. Major earthquakes in Chile and Peru severely damaged the fishmeal and oil plants in those countries. A major oil spill in the Gulf of Mexico has impacted the menhaden fishery in the USA, and volcanic eruptions in Iceland have had some effect on the Scandinavian and European fisheries. Environmental groups are raising concerns about sustainability of the resources. The fish oil industry appears to be in another transition moving from aquaculture to the omega 3 nutritional oil market and prices for both fishmeal and oil in 2010 make the unprecedented 1999 prices look normal. Companies are again looking for alternatives to fishmeal and oil because of these high prices. This paper will focus on several areas to put the industrial fishery into perspective during this latest transition period and draw some comparisons to the previous transition period.


(6) AQUACULTURE WASTE MANAGEMENT – GOING GREEN Cyr Couturier (Canada)

Centre for Aquaculture and Seafood Development, School of Fisheries, Marine Institute, Box 4920, St. John’s, NL Canada A1C 5R3

Farmed sources now account for over 50% of world fish production for human consumption. Aquaculture output is projected to increase 6-9% per annum over the next few decades to meet global seafood demand. Over the same timeframe, capture fisheries production is expected to remain static as it has been for the past three decades. The increase in fisheries output from both extensive and intensive farming sources will continue to generate a variety of pre- and post-harvest waste streams that will need to be dealt with in both economical and environmentally sustainable fashions. The major waste streams are presented in this discussion for pre- and post-harvest operations around the globe, with emphasis on intensive fish production situations. Some of the innovative solutions adopted for rendering wastes into value-added products are discussed. Finally, the focus will be on those technologies that reduce carbon footprint from either waste reduction and / or utilization on farms. By doing so, costs of production are reduced or mitigated, but most importantly efforts to reduce environmental impacts are being developed to ensure the social sustainability of the industry. (7) STEROL CONTENT AND COMPOSITION OF MUSSELS FED DIFFERENT

DIETS Iyad A. Hailat, Christopher C. Parrish, Robert J. Helleur (Canada)

Chemistry Department and Ocean Sciences Center, Memorial University of Newfoundland, St. John’s, NL, Canada A1B 3X7

Dietary sterols known as phytosterols have many health benefits including reduction of the level of cholesterol in blood. One excellent source is mussels. This project examined sterols in cultured blue mussels obtained commercially and from laboratory feeding experiments. Mussels were fed for seven months and the levels of sterols were compared at the beginning and at the end of the experiment. The dietary treatments were feeding on algae, feeding on fish waste in a co-culturing situation, and unfed. Sterols were extracted from whole mussels by a mixture of chloroform/methanol (2:1) and seventeen sterols were identified using GC-MS as their TMS derivative. Six sterols were quantified using GC-MS selected ion monitoring. One way ANOVA and T-tests were used for comparing the levels of sterols. The highest sterol abundance in cultured mussels was cholesterol (321.5), the lowest campesterol (12.6) mg/kg. There were statistically significant differences between the content of sterols in starved mussels and those that either were feeding on algae or fish waste. The concentration of cholesterol, sitosterol and campesterol in waste-fed mussels increased while brassacasterol and desmosterol decreased. The concentration of cholesterol, desmosterol and campesterol in algae-fed mussels increased whereas brassacasterol and sitosterol decreased over the experiment.


(8) BIOCHEMICAL COMPOSITION OF BLUE MUSSELS (MYTILUS EDULIS) FED EFFLUENT FROM AN ONSHORE ATLANTIC COD (GADUS MORHUA) AQUACULTURE FACILITY

A. Both, C.C. Parrish, R. Penney, R.J. Thompson (Canada) Ocean Sciences Centre, Memorial University of Newfoundland, St. John’s, Newfoundland and Labrador, Canada, A1C 5S7

Diet influences the nutritional value of mussels including lipid and fatty acid composition. Given the increasing popularity of integrated multi-trophic aquaculture, which uses extractive organisms (i.e. mussels) to mitigate aquaculture wastes, it is important to know how such practices affect mussel composition. Performance of mussels fed effluent from Gadus morhua was compared to a commercial shellfish diet. Dry weight, ash free dry weight, shell length and condition index of mussels fed algae were significantly higher than in those fed effluent. Effluent fed mussels had a lower acetone mobile polar lipid content (3.0±1.2% total lipid) than algae fed mussels (5.9±1.0% total lipid). There was an increased proportion of the NMID 20:2a and the zooplankton marker 20:1�9 in mussels fed effluent (4.3±0.6 and 4.8±0.2% total FA) compared to those fed algae (2.7±0.3 and 3.2±0.3% total FA). Results suggest effluent is an inferior diet to algae; however, it can supplement growth when the normal diet is scarce. Physical and biochemical characteristics of the effluent diet were also determined. Only 36% of the particles present in effluent are suitable for mussel ingestion; however, particles available for ingestion have the greatest potential to spread to surrounding areas. (9) EFFECT OF DIET AND TEMPERATURE ON MUSCLE LIPID AND FATTY

ACID COMPOSITION IN ADULT AND JUVENILE STEELHEAD TROUT (ONCORHYNCHUS MYKISS)

M.P.A. Wijekoon1, C.C. Parrish1, K. Nag2, A.A.H. Mansour3, D. Whelan4 (Canada) 1Ocean Sciences Centre, 2Department of Biochemistry, Memorial University of Newfoundland, 3Fisheries and Oceans Canada, 4Department of Fisheries and Aquaculture, St John’s, Newfoundland

Fluctuating water temperature is stressful for cultured fish at different life stages, compromising growth, survival and flesh quality. Dietary essential fatty acids play a vital role in marine fish culture. Two studies were conducted to examine the effect of temperature fluctuations on muscle lipid and fatty acid composition in juvenile and adult steelhead trout fed 2 dietary combinations. In study 1, adult fish were fed with 3 commercial diets commonly used by aquaculture farmers. The lipid portion of the 3 diets differed based on origin: marine or terrestrial animal, or vegetable oil. In study 2, juveniles were fed with 3 experimental diets in which the lipids were mainly from fish oil, flax seed or sunflower oil. In both studies the temperature was gradually raised while sampling fish every 12 – 14 days after each increment (study 1: 13.5-18.5°C; study 2: 10.0-18.0°C). The effect of diet on muscle lipids was compared between fish in both experiments, including PUFA and MUFA, DHA:EPA and saturated:unsaturated fatty acid ratios, cholesterol levels and cholesterol to phospholipid ratios. Preliminary results indicate lipid class and fatty acid composition respond to dietary manipulation and temperature changes. These data have implications for fish health and nutritive value for the consumers.


(10) ALTERNATIVE SPECIES IN FISHERIES AND AQUACULTURE Annie Mercier (Canada) Ocean Science Centre, Memorial University, St. John’s, NL, Canada, A1C 5S7 Declines in traditional harvests, shifts in world markets and advances in harvesting technologies are fuelling the development of non-traditional fisheries and productions. In Canada, increasing interest is being expressed for scientific (exploratory) fisheries licenses and many alternative aquaculture species are being considered to enhance natural captures or access new market opportunities. Species under study include seaweeds, sea urchins, abalones, jellyfish, toad crabs and porcupine crabs. Furthermore, sea cucumbers are at the forefront of a new emerging fishery in Atlantic Canada. While sea cucumber harvests date back several centuries in the Indo-Pacific and currently target over 80 edible species, commercial sea cucumber harvests in North America are in their infancy. Differences in biological features, stock abundances, catch efforts and socio-economic importance of the resource are easily identifiable and should be carefully evaluated in light of worldwide experience to develop proper management strategies for this resource. (11) FUTURE FISH PROCESSING TECHNOLOGIES Robert W. Verge (Canada) Canadian Centre for Fisheries Innovation (CCFI), St. John’s, NL, Canada Human needs and wants remain much the same from one generation to another. But how they are met changes as technologies improve over time and competition drives adoption of the new technologies. Fish processing technology has been relatively slow to evolve, for a variety of reasons. Understanding those reasons is key to developing new technologies and new applications of technologies already in use elsewhere. Changing customer expectations, aquaculture, new technologies, and competitive pressures all point toward adoption of some new fish processing technologies in the future.


(12) LIPID CLASS AND FATTY ACID CONTENT AND COMPOSITION OF BIVALVES IN THE VICINITY OF ATLANTIC SALMON FARMS IN COASTAL BRITISH COLUMBIA

E. Marie George, Christopher C. Parrish (Canada) Ocean Sciences Centre, Memorial University, St. John’s, NL, Canada A1C 5S7 Finfish aquaculture enriches the local environment through output of food particles and faecal pellets to the benthos. Analyzing the quality and quantity of material exiting the farms and its uptake by surrounding invertebrates, including edible bivalves, indicates the effectiveness of muti-trophic, co-culturing systems where waste from one species is recycled as food for adjacent species. For this study, molluscs were collected in transects surrounding Atlantic salmon cage sites in Clayoquot Sound and the Broughton Archipelago, BC. Regressions showed blue mussel (Mytilus edulis) wet weight was higher closer to the farms (p=0.0313, n=89). In contrast, mussels had lower proportions of the essential fatty acid DHA closer to the farm (p=0.006). Breakpoint analysis of the mussels showed the zooplankton fatty acid feed marker significantly lower at distances greater than 400 m. In addition, mussel DHA was significantly higher at distances >306 m suggesting an abundance of other, non-essential lipid sources closer to the farm. Principal components analysis showed filter feeders, including mussels and the clam (Protothaca staminea), were associated with higher proportions of PUFA and �3 fatty acids. Mussels also had significantly higher proportions of the available essential fatty acids compared to other molluscs, consistent with their use in integrated mutli-trophic aquaculture. (13) LIFE CYCLE ANALYSIS OF BIOFUELS FROM FISH WASTE FROM REMOTE

FISH PROCESSING PLANTS IN NEWFOUNDLAND AND LABRADOR Punyama Jayasinghe, Dr. Kelly Hawboldt (Canada)

Faculty of Engineering and Applied Science, Memorial University of Newfoundland, St. John’s, NL, Canada

In areas of substantial fish processing such as Newfoundland and Labrador, deriving biofuels from fish waste and use on-site can have advantages such as; reducing the impacts of waste discharge, and reducing emissions and costs for petroleum fuels use and transport. Currently, the waste is sent to landfill and/or discharged to the ocean and depending on the fish species, between 3-25% of the waste is oil. The study is conducted to determine the feasibility and impacts of using fish waste derived biofuel as a blend for use on-site, in the community, or in marine vessels. Waste from three fish processing plants was characterized for chemical composition, stability, reactivity, and partitioning. A process to separate the oil was developed by modifying the fishmeal process. Recovered oil was analyzed for physical properties such as; density, viscosity, melting properties and thermal stability. Chemical composition was analyzed for sulphur content, lipid classes, and fatty acids. Using energy consumption and oil recoverability data for the proposed process, an overall life cycle analysis is conducted. Emission studies were carried out for in-plant use in furnaces, stationary diesel engines and residential boilers. Significant reductions are seen in CO2, SO2, PM and CO, except for NOx emissions.


September 9, 2010 (14) NOVEL FUNCTIONALITY OF MARINE ALLENIC CAROTENOIDS Kazuo Miyashita (Japan) Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611, Japan

Obesity has increased drastically in recent years and is currently considered to be a major risk factor for type-2 diabetes, hypertension, and dyslipidemia. The cluster of these three diseases is called the metabolic syndrome and its incidence is a worldwide problem. The excessive fat accumulation observed in obesity leads to the dysregulation of adipocytokine production in white adipose tissue (WAT), which is closely involved in the development of metabolic syndrome. A few molecular targets offer the most hope for the metabolic syndrome therapeutics. These promising targets have been quite well validated biologically. This presentation introduces describe the protective effect of allenic carotenoids from marine products against metabolic syndrome with special reference to its regulations on the relative gene and protein expressions. (15) VALUE ADDED UTILIZATION OF SEAFOOD PROCESSING BY-PRODUCTS Hordur G. Kristinsson (Iceland)

Acting CEO, Director, Biotechnology and Biomolecule Division, MATIS - Icelandic Food and Biotech R&D, Reykjavik, Iceland

Vast amounts of marine based raw materials are still in large part underutlized. Major opportunities exist with these raw material sources as they are rich in a large variety of natural and highly functional compounds, which with proper extraction, isolation and processing techniques can find use in various foods, specialty feeds, neutraceuticals, cosmeceuticals and even some medical products. The market for natural products is growing very rapidly, particularly products which possess bioactive properties which can have positive effects on health and performance. The past few years have seen significant advances in the isolation and production of novel ingredients from underutilized raw materials. This includes the production of bioactive fish peptides, cartilage compounds such as chondroitin sulfate, glucosamine, prolamine and various seaweed based compounds, to name a few. Some of these ingredients have very unique functions compared to their non-marine counterparts, and display very high activity. The industry is realizing that very significant value addition can be realized with underutlized raw materials. However for this industry to become successful and compete in the marketplace, patience is needed as well as continued support to research and development. Particular attention to marketing strategies is also important for these ingredients to stand out in the marketplace. �


(16) EXTRACTION AND PROCESSING OF OMEGA-3 OIL FROM MICROBIAL BIOMASS

Udaya N. Wanasundara (Canada) Senior Scientist and Project Leader, POS Pilot Plant Corporation, 118 Veterinary Road, Saskatoon, SK, Canada, S7N 2R4

There has been a considerable interest in the use of microbial biomass as feed stocks for production of high value lipids. Marine algae typically have a high content of long-chain omega-3 polyunsaturated fatty acids (PUFA) considered to be a valuable biomass to extract high PUFA-lipids. Algae oils destined for food and nutraceuticals or natural health products often require unique procedures aimed at minimizing the losses of valuable components during extraction and further processing. These procedures include special extraction and oil processing methods different from other plant feed stocks. This presentation will provide an overview of procedures that are employed in extracting and further processing of oil from algal sources.

(17) UNDERSTANDING THE IMPACT OF MARINE MAMMALS ON FISH RESOURCES IN ATLANTIC CANADA

Garry Stenson (Canada) Science Branch, Department of Fisheries and Oceans, St. John’s, NL,Canada The impact of marine mammals, particularly seals, on the recovery of depleted fish stocks is a controversial issue and the focus of significant research efforts. Three species of seals are considered important predators in the northwest Atlantic, harp, hooded and grey seals. Consumption of important prey species by seals has been estimated using bioenergetics models and a number of studies have attempted to determine the impact of seals on fish stocks, particularly the impact of harp and/or grey seals on Atlantic cod. In general, these studies have indicated that although seals consume substantial amounts of commercial fish species and important forage species, the impact of these removals on the current fish stocks is difficult to determine. Seals are important predators of both large and small cod and could be playing a role in the non-recovery of cod stocks, but seal predation can not account for a large component of mortality in most areas and therefore, the total impact of seal predation cannot be determined. Although less is known about the impact of whales, preliminary consumption estimates suggest that they may also be important predators.


(18) FISH PROTEIN HYDROLYSATES FROM COD/SALMON THAT COULD BE PROMISING FOR ANTI-DIABETIC FUNCTIONAL FOODS

Fida Hasan1, Andre Marette2, Tom A. Gill1 (Canada) 1Canadian Institute of Fisheries Technology, Dalhousie University, Halifax, Nova Scotia, B3J 2X4; 2INAF, Laval University, Quebec City, G1V 4G2

Recently published articles (V´eronique et al., Diabetes Care 30:2816–2821, 2007; Picot et al., Process Biochemistry 41:1217–1222, 2006; Gildberg et al., Comp. Biochem. Physiol. l14B:97-101, 1996; Tremblay et al., Annu. Rev. Nutr. 27:293–310, 2007; Lavigne et al., Am J Physiol Endocrinol Metab 281: E62–E71, 2001) suggest that proteins from cod/salmon may be promising for the development of functional foods and nutriceuticals. However, there have been only limited scientific studies regarding the true biological activity of fish proteins compared to the well known benefits provided from ingestion of omega-3 lipids found in fatty fish species. Since fish proteins are far too large to be transported across the intestinal mucosa, it was logical for us to begin our work with fish protein hydrolyzates (FPHs) produced in vitro using artificial digestive juices. Fresh farm-raised salmon and cod frames were provided by Cooke Aquaculture Inc., Saint John, NB (industry research partner). The cod/salmon frames were deboned with a Bibun Model SDX-16, meat/bone separator equipped with a drum containing 5 mm diameter holes (Bibun Manufacturing, Hiroshima, Japan). FPHs were made from deboned minced cod and salmon treated with one of four proteases (Alcalase, Pepsin, Pancreatin and Papain) or by sequential digestion with Pepsin followed by Trypsin and Chymotrypsin. Another sequential digestion (Pepsin followed by Pancreatin) was also tested. Crude fractionation of the FPHs was carried out in a Millipore Prep/Scale spiral wound ultrafiltration module (Millipore Inc., Nepean, ON, Canada) equipped with a 10 kDa, a 5 kDa or a 1 kDa TFF membrane. Molecular weight distributions of cod/salmon hydrolysate peptides were subsequently determined for each fraction from each FPH using size exclusion chromatography in FPLC mode on a Superdex Peptide HR 10/300 column (AKTA FPLC; GE Healthcare). The FPLC fractions were subsequently analyzed by Reverse Phase-HPLC using gradient solvent system using C18 column. The reproducibility of protein solubilization and peptide production was confirmed by SDS-PAGE and RP-HPLC peptide elution profile. RP-HPLC elution profiles from various digestions confirmed that a considerable number of low molecular weight peptides (< 1000 Da) were produced. Anti-diabetic activities of individual freeze-dried fractions are currently being measured on cell lines grown in vitro (Laval University).


(19) FISH PROTAMINE (PTM), A PROMISING ANTIBACTERIAL BIOACTIVE: MECHANISM OF INTERNALIZATION THROUGH OUTER MEMBRANE CHANNEL PROTEINS (PORINS) OF ESCHERICHIA COLI AND OTHER GRAM-NEGATIVE BACTERIA

Hasan M. Fida1, Mukund Mohan, Mathias Winterhalter2 and Tom A. Gill1 (Canada) 1Canadian Institute of Fisheries Technology, Dalhousie University, P.O. Box 1000, Halifax, Nova Scotia. [email protected] 2School of Engineering and Science, Jacobs University Bremen, D-28759, Bremen, Germany

Protamine (Ptm), is a cationic antimicrobial peptide (CAP) derived from fish milt (spermatic cells) and used in medical applications as a carrier for injectable insulin, a heparin antagonist and more recently as an antibacterial ingredient in some food products. It is an arginine-rich CAP (Mr 4-4.2 kD) with a pI of 11-13. It is well known that Ptm has no stable secondary structure due to the even distribution of positive charges along the peptide backbone. Ptm is perhaps an unusual CAP in that it appears to be able to internalize certain Gram-negative bacteria without forming permanent pores or causing cell lysis. The outer membranes of Gram-negative bacteria contain barrel-shaped porins for the passage of hydrophilic solutes, basic amino acids and small peptides less than 650 Da. This study was concerned with the mechanism of Ptm internalization. The patch clamp experiments on synthetic bacterial membranes confirm that Ptm does not bind to DPhPC (phospolipid) bilayers, and therefore will not likely internalize via poration. Furthermore, we have shown with computer modeling studies that the probability of Ptm crossing a porin-free membrane (DPhPC) is very low. Upon insertion of the purified porins OmpF, OmpD and OprF isolated from E. coli 29522, S. typhimurium 14028 and P. aeruginosa PAO1 27853, respectively, we found that Ptm can translocate through an outer membrane via a porin. Single channel recordings suggested that OmpF OmpD and OprF porin channels were strongly blocked and unblocked for defined periods of time, suggesting translocation. Translocation of Ptm inside the channel caused fluctuations in the ion current, reflecting the molecular interactions with the channel wall. The study was complemented with computer based molecular modeling and immunoelectron microscopy and we show here for the first time that CAPs may be internalized via porin channels without causing substantial membrane disruption, poration or lysis.


(20) PROTAMINE INTERACTING WITH INNER MEMBRANES OF GRAM-NEGATIVE BACTERIA

David A. Pink, Bonnie Quinn (Canada) Physics Department, St. Francis Xavier University, Antigonish, NS Canada B2G 2W5 A minimal (“coarse-grained”) model has been created to study the interactions of the CAP protamine (Ptm), with the inner membranes of Gram-negative bacteria. The intent is to establish the extent to which Ptm is bound to these membranes with a view to predicting whether Ptm penetrates these membranes via pores or other defects in the membrane or whether some other mechanism for internalization is required. The inner membrane is represented by a phospholipid bilayer composed of phosphatidyl-ethanolamine and –glycerol. We include screened electrostatics and hydrogen bonding. We represent monovalent ions in the aqueous solution by a combination of implicit ions, described by a Debye screening length, and explicit ions represented by charged spheres. All divalent ions are represented by charged spheres. We found that (i) Ptm does not get bound to the inner membrane and predict that (ii) excess Calcium ions will bring about an increase in the MIC and MBC. Some of the experiments proposed have been carried out and we shall outline further experiments to be done. Work supported by AFNnet-NCE, NSERC of Canada and ACENET. (21) SUPERCRITICAL FLUID EXTRACTION AND CHROMATOGRAPHY OF

MARINE BIOACTIVES Isabelle Vu Trieu (USA) Waters Corporation, Milford, USA This presentation will cover the fundamentals of Supercritical Fluid Extraction (SFE) and Supercritical Fluid Chromatography (SFC) and their applicability for the extraction and analysis of bioactives from marine biomass. In SFC and SFE we use supercritical CO2 as the extraction solvent or the chromatography mobile phase. The advantages are little or no residual solvent, superior purity and yield, faster separations, low operations costs, scalability (from the laboratory to the production plant), compatibility with thermally labile products, increased safety and environmental friendliness compared to solvent-based systems. SFC and SFE are green technologies that can significantly reduce or eliminate organic solvent use and offer recycling capabilities. We will describe some of the key physical properties of supercritical CO2 and their implication on extraction and separation. Typical hardware and SFC detection schemes will be presented. Application examples will illustrate the talk, and will include the SFE of natural products from marine biomass, SFC of biodiesels and SFC of fish oil. Cost benefits and the “green” advantage will be discussed.


(22) DETECTING TAINT IN SEAFOOD DURING THE GULF OF MEXICO OIL SPILL

Lucina E. Lampila (USA) Seafood Technology, Louisiana State University of Food Science, Baton Rouge, LA, USA 70803

There is a great deal of concern over the safety of seafood since the explosion of the Deepwater Horizon drilling platform and subsequent oil spill. Rather than posting signs indicating that they are proudly serving Louisiana seafood, restaurants are posting signs stating that they are not serving Louisiana seafood. Negative media attention is perpetuating this misconception. The first line of defense to detect if seafood has been contaminated by oil is olfactory. Oil contains polycyclic aromatic hydrocarbons (PAHs) which impart a distinctive odor. Field screeners are trained to detect the presence of PAHs to 10 ppm while Expert screeners have sensitivity to 0.5 ppm. Rapid screening is critical due to the size of landings in the gulf and analytical screening is expensive with relatively few facilities able to conduct the official method of analysis. This presentation will focus on the screening process by sensory evaluation. (23) EFFECT OF ENZYMATIC RANDOMIZATION ON POSITIONAL

DISTRIBUTION AND STABILITY OF OMEGA-3 OIL TRIACYLGLYCEROLS Jiankang Wang, Fereidoon Shahidi (Canada) Department of Biochemistry, Memorial University, St. John’s, NL Canada A1B 3X9 Randomization has been commonly used to modify chemical and physical properties of natural fats and oils. In this study, seal blubber oil (SBO) and menhaden oil (MHO) were modified through enzymatic randomization using Novozyme 435 and the effect on positional distribution of fatty acids was investigated using gas chromatography (GC). The effect of randomization on the stability of the original oils and their randomized counterparts was analyzed by comparing conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS) values after accelerated oxidation at 60 �C for 4 days. The data obtained indicated that polyunsaturated fatty acids were redistributed more evenly among the terminal (sn-1, sn-3) and middle (sn-2) positions in the randomized oils when compared with their unrandomized counterparts. The stability of the randomized oils was also affected to different degrees due to redistribution of fatty acids and changes of �-tocopherol content.


(24) ENZYMATIC SYNTHESIS OF PHYTOSTEROLS ESTERS WITH OMEGA-3 PUFA FROM MARINE SOURCES

Zhuliang Tan, Fereidoon Shahidi (Canada) Department of Biochemistry, Memorial University, St. John’s, NL, Canada, A1B 3X9 Phytosteols occur in the cell membranes of plants. They are present at low levels in grains, fruits, vegetables as well as marine algae. High content of phytosterols or their ester derivatives (60-80%) have also been reported in surf clam, Greenland cockle, blue mussel, and icelandic scallop. In addition to their well-known effect in lowering low-density lipoprotein (LDL) cholesterol, phytosterols possess other biological activities, such as anti-inflammatory, anti-antherogenicity activity, and anti-cancer potential. Long-chain polyunsaturated fatty acid (PUFA), especially omega-3 PUFA, have been demonstrated to possess cardioprotective and immune-enhancing effects. Esterification of phytosterols with omega-3 PUFA may render better physiochemical properties such as solubility, miscibility, oxidative stability and hence bioactivity and bioavailability. Thus, phytosterol esters with omega-3 PUFA may offer both the benefits of phytosterol and those of long-chain omega-3 PUFA, possibly in a synergistic manner. This contribution describes a method for enzymatic synthesis of phytosterol esters of omega-3 PUFA from marine sources. Keywords: Phytostreols, Omega-3 PUFA, Enzymatic Synthesis (25) EXPLORATORY FISHING SURVEYS AND BIOLOGICAL RESOURCE

ASSESSMENT OF ATLANTIC HAGFISH (MYXINE GLUTINOSA) IN THE NEWFOUNDLAND REGION

S.M. Grant, W. Hiscock, G. Bishop, R. Sullivan (Canada) Marine Institute of Memorial University, St. John’s, NL, Canada A1C 5R3 Hagfish fisheries the world over have generally been characterized as boom and bust, which may in part be attributed to a lack of sound science in advance of commercial development. Exploratory Atlantic hagfish (Myxine glutinosa) fisheries conducted in the Newfoundland region have provided valuable information on the relative abundance, distribution, and biology of this underutilized resource as well as determined the selective properties of baited traps. Hagfish are concentrated in the warmest available bottom temperatures, females dominate (>98%) the commercial catches, and length at maturity varied geographically. Low reproductive potential indicates a high susceptibility to over exploitation. Juveniles and undersized Atlantic hagfish were found to be highly susceptible to capture in baited traps. This study demonstrates the importance of cooperation between science and industry and the development of a well laid out research plan when investigating the commercial potential of underutilized species.


(26) RECOVERY AND CHARACTERIZATION OF A SERINE COLLAGENOLYTIC EXTRACT FROM SNOW CRAB (CHIONOECETES OPILIO) BY-PRODUCTS

Nathalie Souchet, Serge Laplante, Piotr Bryl (Canada) Centre technologique des produits aquatiques, 96, mont�e de Sandy Beach, Bureau 1, 07, Gasp�, Quebec, Canada G4X 2V6

Sequential acidic precipitation followed by a single chromatographic step (gel filtration) allowed the recovery of a collagenolytic fraction containing several proteases from snow crab (Chionoecetes opilio) by-products. The partial purification was particularly efficient to recover trypsic (purification fold = 1352.5; yield = 110%) but also chymotrypsic, elastolytic and collagenolytic activities. A temperature of 40°C and pH 8.0-8.5 were optimal for enzyme activity, which was stable for two hours under these conditions. Calcium was not required for stability. The isoelectric points of the protein components ranged from 3.7 to 4.6. Zymography revealed a 29-kDa major component and others between 22 kDa and 56 kDa. Enzymes were inhibited by PMSF and TCLK but were insensitive to TPCK. Inhibition by EDTA could be due to another mechanism than Ca2+ chelation. In view of these properties, the proteases likely belong to the serine collagenase group. Using a food system (ground herring), the fraction was more proteolytic than a commercial bacterial protease (Protamex®), suggesting potential applications in enzymatic hydrolysis processes on marine by-products. (27) ENHANCING SUSTAINABLE MUSSEL INDUSTRY PRODUCTION AND

GROWTH THROUGH ASSESSMENT AND REMOVAL OF CONSTRAINTS IN SEED SUPPLY Tracy Granter1, Christopher Dawe1, Kiley Best1, Thomas Brown1 and Darrell Green2 (Canada) 1Centre for Aquaculture and Seafood Development, Marine Institute of Memorial University of Newfoundland, St. John’s, NL A1C 5R3, Canada; 2Newfoundland Aquaculture Industry Association, 11 Austin Street, Suite 209, St. John's, NL, A1B 4C1, Canada

The Newfoundland Aquaculture Industry Association (NAIA) requested the Marine Institute’s Centre for Aquaculture and Seafood Development’s (CASD) assistance to evaluate the amount and the performance of mussel seed collected from traditional and new collection sites in Placentia Bay, Bonavista Bay and Notre Dame Bay over the past 5 years. The objective was to determine each site’s potential as a source of quality mussel seed to aid future mussel aquaculture industry development. Sampling sites were selected by a steering committee each year, larval monitoring was completed at each of the sites and experimental seed collectors were deployed at the selected sites. Oceanographic data and seed samples were collected in the spring and fall for each site and morphometric and genetic analysis were performed on each seed sample. Seed transfers were also completed from Bonavista Bay and Notre Dame Bay donor sites and transferred to a recipient mussel farm in Placentia Bay. This project has been supported by NRC-IRAP, the Department of Fisheries and Oceans (ACRDP), Department of Fisheries and Aquaculture, the Canadian Center for Fisheries Innovation (CCFI) and Newfoundland mussel industry partners. Memorial University of Newfoundland’s biology department provided the expertise for the mussel seed genetic analysis.


(28) DEVELOPMENT OF AN UNDERWATER CAMERA SYSTEM TO OBSERVE THE HARVESTING TECHNIQUES OF COMMERCIAL SPECIES

Melanie Underwood, Paul Winger, George Legge (Canada) Fisheries and Marine Institute, Memorial University, P.O. Box 4920, St. John’s, NL, A1C 5R3

The commercial fishery is an expensive industry and requires a multitude of resources to move the product from the ocean to the consumer. Fuel costs and unintentional species caught in harvesting gear have come under considerable scrutiny in recent years, encouraging harvesting gear to become more efficient and species selective. However, prior to any gear modifications, a better understanding of the species behaviour and interaction to the harvesting gear is required. The use of an underwater camera system to quantify species behaviour in relation to capture gear is a relatively new application to the fishery, and has only recently been developed to incorporate high definition (HD 1080i/720p) digital technology. Inspired by cameras used in remotely operated vehicles (ROVs), the system designed for this research was developed to observe fish behaviour, and is ideal for capturing objects in motion as they relate to operating trawls. This type of research has previously led to new gear designs that have reduced the quantity of non-targeted and undersized species often caught in commercial fisheries. The camera system developed is not limited to trawl research but can be applied to stationary gear, such as pots or traps, and other forms of mobile gear.


September 10 (29) UTILIZATION OF MARINE OILS IN FOOD AND NON-FOOD APPLICATIONS Roland Verhe1, V. Van Hoed1, C. Echim1,2, W. De Greyt2 (Belgium) 1Ghent University, Belgium; 2De Smet Ballestra, Zaventem, Belgium The beneficial health effects of marine oils containing high amounts of omega-3 polyunsaturated fatty acids have attracted the interest of the public for the use in nutraceuticals. However, marine oils have to undergo a series of refining steps in order to be used in food products. Refining involves degumming for the removal of phospholipids, a bleaching step and deodorization for the elimination of oxidation products and free fatty acids. Due to the high unsaturation , conventional refining processes have to be adapted in order to guarantee a high concentration of the omega-3 fatty acids. In addition, marine oils can be contaminated by dioxins, benzofurans and polychlorinated biphenyls. A process has been developed for the removal of the contaminants involving a combined adsorption with active carbon followed by stripping at 190 °C in vacuum. Marine oils not suitable for human consumption and animal feed can be used as a feedstock for the production of biodiesel using a transesterification producing either methyl or ethyl esters. A comparison of biodiesel produced from marine oils and waste frying oils will be made. Generally, the quality of biodiesel from both resources is similar but biodiesel from marine oils suffers from a low oxidative stability. An alternative use of marine oils is the production of a biofuels for the generation of power and heat which is an excellent feedstock for green electricity. (30) PROTEIN AND LIPID OXIDATION IN FISH AND FISH PRODUCTS Caroline P. Baron (Denmark)

Division of Industrial Food Research, Technical University of Denmark, National Food Institute, Søltofts Plads, Building 221, DK.-2800 Kgs Lyngby, Denmark

Most of the research on oxidation in fish and fish products has been done on the oil fraction probably because their oxidation is immediately noticeable as it results in the formation of significant off-flavour and gives a characteristic rancid taste. In contrast, protein oxidation has only received some attention very recently. Perhaps, this is because oxidation of food proteins results in more subtle changes that are not immediately perceives by the human senses but also due to the complexity of the reactions involved. Compared to lipids, proteins are very large and complex molecules organised in large structures and oxidation may lead to a great number of protein modifications. This presentation will focus on protein oxidation and present the current understanding of protein oxidation in foods. In addition, the importance of protein oxidation in fish and fish products will be demonstrated by taking two examples from our research. The first example will deal with protein and lipid oxidation in frozen rainbow trout and the second example will deal with protein and lipid oxidation in salted herring. A more fundamental understanding of the mechanism is still needed but protein oxidation in fish and fish products should not be overlooked.


(31) USE OF BIOREACTOR FOR PRODUCTION OF MARINE NUTRACEUTICALS Se-Kwon Kim (Korea)

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

Many bioactive substances, such as antitumor, anti-inflammation, anti-allergy, antioxidant, antifungal, anti-HIV and antihypertensive, have been identified and investigated from marine organisms using marine biotechnology. Recently, because of the land pollution and limitation in new material development using inland organisms, there has been an increasing interest on utilization of marine organisms and marine biotechnology for development of bioactive substances which can be used as nutraceuticals. Moreover, bioconversion of unutilized byproducts to valuable nutraceuticals using marine biotechnology also has a great potential in marine bioprocess industry. With respect to investigate and develop marine nutraceuticals, many studies have been conducted to develop various biotechnologies, such as membrane bioreactor, bioconversion and continuous mass producing process technology and etc. Membrane bioreactor equipped with ultrafiltration membranes is recently emerging bioconversion technology for the development of nutraceuticals and considered as a potential method to utilize marine organisms and byproducts efficiently.

(32) VITELLOGENIN FOR DETERMINATION OF FISH SEXUAL AND REPRODUCTIVE MATURATION AND PRESENCE OF XENOBIOTICS

Joseph H. Banoub (Canada) Fisheries and Oceans Canada, Science Branch, Environmental Sciences Division, Special Projects, P.O. Box 5667, St John’s, Newfoundland, A1C 5X1 and Department of Chemistry, Memorial University, St. John’s, Newfoundland, A1B 3V6, Canada

Vitellogenin is a complex phosphoglycolipoprotein that is secreted into the bloodstream of sexually mature, female, oviparous animals in response to circulating estrogens. It is then incorporated into the ovaries by receptor mediated endocytosis, where it is further cleaved to form the major constituents of the egg yolk proteins. It is generally accepted that these protein and peptide products serve as the main nutritional reserve for the developing embryo. Quantification of vitellogenin in blood is useful for different purposes. The reproductive status and degree of sexual maturation of oviparous animals can be assessed according to the levels of vitellogenin in plasma. The expression of this protein can also be induced in males under the effect of estrogenic compounds. Relying on this observation, vitellogenin has been used as a unique biomarker of environmental endocrine disruption in many species. In this respect, vitellogenin levels could potentially be used to assess the estrogenic activity. In this conference, we review a technique developed in our laboratories for measuring vitellogenin plasma levels of different fish species using high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry.


(33) THE USE OF HIGH PRESSURE TREATMENT FOR INCREASING THE SHELF-LIFE OF SEAFOOD-BASED CHOWDER Malco C. Cruz-Romero1, R. Erdilal2, C. Legrand1, D. Gilroy1, J.P. Kerry1 (Ireland) 1School of Food & Nutritional Sciences, Food Packaging Group, University College Cork, Cork, Ireland; 2Faculty of Fisheries, Akdeniz University, Antalya, Turkey and 3Department of Biological Sciences, Cork Institute of Technology, Cork, Ireland

High-pressure (HP) processing, a ‘residue-free’ preservation technique, is a physical method that can be used for cook-chill meals like chowder in enhancing their food safety design. The objectives of this research were to study the effects of HP treatment at 300, 400 or 500 MPa for 5 min on the microbiological and physicochemical changes associated with chowder stored under vacuum packaging conditions at 4°C. After HP treatment chowder had significantly (P < 0.05) higher pH, lipid oxidation (TBARS) and CIE L-values compared to untreated samples; values being dependant on the pressure of the treatment applied. During storage, TBARS and total volatile base nitrogen (TVBN) values increased significantly (P < 0.05) in untreated samples; however TBAR’S and TVBN values did not increase significantly in HP-treated samples. From a microbiological perspective, immediately after HP treatment, the microbiological load was below the detection limit and HP delayed subsequently microbial growth compared to untreated samples. In samples HP-treated at 400 or 500 MPa, Staphylococcus aureus, Bacillus cereus and E. coli and coliforms were absent throughout product storage, with LAB being the dominant spoilage microflora. The shelf-life of chowder HP-treated at 500 MPa was extended by 28 days compared to the untreated sample. HP treatment, in combination with adequate chilled storage, can extend the shelf-life and increase the safety of seafood-based chowder. (34) THE INTERACTIVE CENTER FOR FISH (“CENTRE INTERACTIU DEL

PEIX”, CIP) IN BARCELONA, SPAIN Reyes Pla (Spain) Tecnologia dels aliments, Departament de ciència animal i dels aliments, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain

The Guild of Fish and Seafood Wholesalers of Barcelona was started in 1978. Currently, Guild membership includes all of the companies operating in the city’s Central Fish Market (Mercabarna). The idea of promoting the development of the Interactive Center for Fish (“Centre Interactiu del Peix”, CIP) arose within the Guild’s proactive Marketing Committee, whose mission is to make the Central Fish Market, a modern market for now and into the future, tailored to the 21st century. Currently, approximately 2,000 fishmongers’ shops, smaller municipal markets and supermarkets, and restaurants buy fish products at this market, which serves some 4,000,000 consumers in the great Barcelona area. The CIP wants to be the focal point in Catalonia for everything related to fish as food, including developing 6 exhibition areas. The main objectives of these exhibits are to familiarize children with fish as a healthy and tasty food, to encourage the consumption of fish by the adult population, to advocate on behalf of the fishing industry and to communicate information about the use of fishery products, to help with the promotion of the commercial structures associated with the industry, to support the inclusion and use of fish in restaurants and foodservice, and to help with research and development related to fish such as new products, new methods of handling and preservation and better presentation.


(35) DEVELOPMENT OF A FISH OIL DERIVED BIODIESEL PRODUCTION SYSTEM FOR NEWFOUNDLAND AND LABRADOR Heather Manuel, Wade Murphy (Canada) Centre for Aquaculture and Seafood Development, Marine Institute of Memorial University of Newfoundland, St. John’s, NL A1C 5R3, Canada

The Centre for Aquaculture and Seafood Development (CASD), in partnership with Memorial University’s (MUN) Faculty of Engineering and Applied Science, has completed year 1 of a 3 year research program to advance current biodiesel conversion technologies for specific applications to marine waste oil feedstocks. The main objective of the current research program is to develop an economically viable, fish oil derived biodiesel production system for rural communities in Newfoundland and Labrador. The major research activities being undertaken include: The development of simple and effective methods to produce biodiesel from waste fish oils; Establishing a pilot scale fish oil biodiesel demonstration system at the Marine Institute’s Atlantic Canada Fisheries and Aquaculture By-Products Research Facility; To design a process to optimize biodiesel yields from waste fish oils; and to technically evaluate the performance of fish oil biodiesel for use in diesel engines/boilers/generators, etc. The focus of year 1 for the CASD was to establish a pilot scale biodiesel demonstration system at the Fishery By-product Research Facility. Funding was provided through the Fisheries Technology and New Opportunities Program (FTNOP) and the Canadian Centre for Fisheries Innovation (CCFI) to acquire a customized mobile 55 gallon batch biodiesel processor. To test the system three production runs were conducted using 570 L of spent vegetable oil provided by Country Ribbon. A yield of 82.5% of vegetable derived biodiesel was obtained. Seaward farms provided 600 L of cod liver oil for this study. However, further pre-processing steps including filtration and an acid pre-treatment are required before it can be converted to biodiesel.




POSTERS (P1) SPECTROSCOPIC AND CHEMICAL INVESTIGATION ON NATURAL

ASTAXANTHIN-RICH-SHRIMP OIL Nad�jda Tchoukanova, Subramanian Balaji, Claude Pelltier, Mathieu Ferron, Yahia Djaoued Institut de Recherche sur les Zones – Moncton University Shippagand, NB, Canada (P2) A NOVEL METHOD FOR RAPID DETECTION OF SODIUM

TROPOLYPHOSPHATE IN PENAEUS AZTECUS AND SETIFERUS SHRIMP Alisa Todd Department of Food Science, Louisiana State University, Baton Rouge, LA 70803 USA (P3) ENZYMATIC HYDROLYSIS OF TWO PLAGIC SPECIES: COMPOSITION OF

LIPID RECOVERY AND DISTRIBUTION Marie-Elise Carbonneau, Piotry Bryl, Serge Laplante Centre Technologique des Produits Aquatiques, Gaspe, Quebec, Canada (P4) THE EFFECT OF FENTON’S REACTANTS AND ALDEHYDES ON THE

CHANGES OF MYOGLOBIN FOR EASTERN LITTLE TUNA (EUTHYNNUS AFFINIS) DARK MUSCLE

Yaowapa Thiansilakul1, S. Benjakul1, M.P. Richards2 1Prince of Songkla University, Hat Yai, Thailand and 2University of Wisconsin, Madison, USA


POSTER ABSTRACTS (P1) SPECTROSCOPIC AND CHEMICAL INVESTIGATION ON NATURAL

ASTAXANTHIN-RICH-SHRIMP OIL Nad�jda Tchoukanova, Subramanian Balaji, Claude Pelltier, Mathieu Ferron, Yahia Djaoued (Canada) Institut de Recherche sur les Zones – Moncton University Shippagand, NB, Canada Astaxanthin (ASTX) (3,3’-dihydroxy-��’-carotene-4,4’-dione) is a ketocarotenoid pigment widely found naturally in some type of microorganisms such as algae, as well as in marine habitants such as shrimps, lobsters etc. It is very well known for its antioxidative activity, biological function as vitamin A precursor, and immunomodulating activities. The Coastal Zones Research Institute in collaboration with the Island Fishermen Cooperative Association Ltd (IFCA) developed a new technology to recover astaxanthin-rich shrimp oil. Our objective is to investigate the recovered astaxanthin-rich shrimp oil obtained during the processing of shrimps. This study gains significance as ASTX is known to exhibit prolonged antioxidant functionality in the presence of fatty acids.

We developed a standard protocol to study astaxanthin-rich shrimp oil by confocal micro-Raman microscope combined with a FTIR-micro analysis system. The molecular structure of ASTX has 9 conjugated bonds attached to the hexagonal rings on either side each having one hydroxyl and ketone groups at 3,3’ and 4,4’ positions, with alkyl groups attached to 9,9’,13,13’ positions. In general, Raman spectrum exhibits strong bands at 1514 cm-1 due to C=C conjugated bonds and C-C stretching vibrations at 1158 cm-1, and the CH wagging modes is seen in the 600 - 800 cm-1 interval. Raman spectrum of �-Carotene, which is isostructural with ASTX, exhibits cis-isomeric forms by showing a significant blue shift to the �(C=C) modes. By comparing the obtained Raman spectra of astaxanthin rich shrimp oil with that of �-carotene, we could identify the trans and cis isomers of ASTX in shrimp oil. IR spectra show modes that are complimentary to Raman spectra. Further they evidence the presence of astx in esterified form, supporting the Raman data. The chemical characterization of the recovered astaxanthin-rich shrimp oil showed a rich profile for fatty-acids and is proven to have very important anti-oxidant activity. These results indicate that, ASTX rich shrimp oil has a potential to exhibit prolonged antioxidant activity. Further Raman and IR spectral features can be used as finger prints to study natural ASTX present in the shrimp oil recovered during the processing of shrimps


(P2) A NOVEL METHOD FOR RAPID DETECTION OF SODIUM TROPOLYPHOSPHATE IN PENAEUS AZTECUS AND SETIFERUS SHRIMP

Alisa Todd (USA) Department of Food Science, Louisiana State University, Baton Rouge, LA 70803 USA Species substitution, mislabeling, undercounting, and over-treating with sodium tripolyphosphate to produce excess water weight have become major concerns in the seafood industry. The objectives of this study are to determine a nondestructive and rapid method of phosphorus detection in shrimp treated with sodium tripolyphosphate at different concentrations and time intervals utilizing NIR spectroscopy. Wild caught Penaeus setiferus and aztecus Louisiana gulf coast shrimp were submerged in deionized water, 1% sodium chloride (NaCl) and 1% sodium tripolyphosphate (STPP), 2% sodium chloride and 2% sodium tripolyphosphate, 2.5% sodium tripolyphosphate, and 5% sodium tripolyphosphate solutions for 30, 60, 120, 240, 480, 960, 1920 minute time intervals. The total moisture was determined and correlated with the water peaks at 2840, 3260, 5587, and 7100 nm on the resulting NIR spectra. Mineral analyses were performed by Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) for the total phosphorus allowed comparison with the baseline spectra at 1908 nm with the spectra-structure of P-OH. There was a detectable shift in the spectra of the treated and control shrimp, which is being investigated further to fully determine the application of NIR technology as a rapid detector for the presence of sodium tripolyphosphate in shrimp. (P3) ENZYMATIC HYDROLYSIS OF TWO PLAGIC SPECIES: COMPOSITION OF

LIPID RECOVERY AND DISTRIBUTION Marie-Elise Carbonneau, Piotry Bryl, Serge Laplante (Canada) Centre Technologique des Produits Aquatiques, Gaspe, Quebec, Canada Two pelagic species, mackerel (Scomber scombrus) and herring (Clupea harengus), were going through the same pilot-scale enzymatic hydrolysis process using Protamex®, followed by various fractionation steps. A great difference in oil extraction efficiency and lipid global distribution was obtained between both fish species. A recovery rate of 84.1% oil was obtained with mackerel, whereas only 50.6% with herring. In the latter case, a higher part of lipids (14,3%) was retained from solids after decantation (the first fractionation step). Oils extracted from mackerel and herring contained respectively 91.5% and 94.8% of triglycerides. From biomass to oil extracted, the omega-3 fatty acid content in mackerel slightly decreased from 24.1% to 23.1%. With herring, a more important decrease was observed, from an already lower initial content of 17.9% to 12.9%. Similarly, the phospholipids concentration was passing from 3.0% to 1.3% (mackerel) and from 2.6% to 0.5% (herring). Those results reveal that enzymatic hydrolysis process conditions were more convenient for mackerel to recover an oil extract with better nutritional qualities, without using high temperatures or solvent. However, process optimization based on the type of enzyme and period of hydrolysis would improve the extraction of herring oil and its nutritional qualities.


(P4) THE EFFECT OF FENTON’S REACTANTS AND ALDEHYDES ON THE CHANGES OF MYOGLOBIN FOR EASTERN LITTLE TUNA (EUTHYNNUS AFFINIS) DARK MUSCLE

Yaowapa Thiansilakul1, Soottawat. Benjakul1, Mark.P. Richards2 (Thailand) 1Department of Food Technology, Faculty of Agro-Industry Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand 2Meat Science and Muscle Biology Laboratory, Department of Animal Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA

The changes of oxymyoglobin and metmyoglobin from Eastern little tuna (Euthynnus affinis) dark muscle as affected by Fenton’s reactants (H2O2 and FeCl2) and aldehydes (hexanal and hexenal) were studied. H2O2, could induce the formation of ferrylmyoglobin. In the presence of Fe2+ and/or H2O2, the changes in fluorescent intensity of myoglobin were noticeable, but there were no changes in aggregation ratio. The oxidation of oxymyoglobin and conformational changes of globin were more pronounced when aldehydes were incorporated. Hexenal had a great impact on cross-linking of oxymyoglobin and metmyoglobin via covalent modification. Alteration of myoglobin redox state might be enhanced by conformational changes of globin induced by both Fenton’s reactants and aldehydes. Keywords: myoglobin; ferrylmyoglobin; H2O2; cross-linking; hexanal; hexenal


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