UJNR Aquaculture Panel

The 37th Scientific Symposium

October 28th – 29th, 2008
Yokohama City Port Opening Memorial Hall
Kanagawa 231-0005, Japan

Aquafeed and its future aspects
Aim of the symposium
Further development of aquaculture is highly desired as a production source ofimportant foods for human being all over the world. Stable supplies of safe andeconomical feed ingredients are requisite for the sustainable development of aquaculture.
Nevertheless, in recent years we recognized some serious problems that threatening theuse of important feed ingredients like fish meal and oil have newly appeared as ourmutual areas of future research.
On this symposium we UJNR aquaculture panel members discuss about the
problems deal with aquaculture feeds and attempt to explore the effective means to
overcome these constrains for the further sustainable advancement of aquaculture.

Program
Tuesday, October 28th, 2008

Registration

Opening remarks (Hiroshi Nakano, Japan Panel Chair)
The Aim of this symposium (Makoto Yamasaki)

9:00-9:30
9:30-9:35
9:35-9:40

Session 1: Current status surrounding aquafeeds and feed research(Moderators: Michael Rust and Heisuke Nakagawa)1. Introduction: Present status of aquafeed research in Japan
9:40-10:00
Takao Yoshimatsu (Japan)
2. The issues surrounding aquaculture feed and the current measures against themin Japan
10:00-10:25
Masahide Fujise (Japan)
13. Challenges for development of aquafeeds
Makoto Nakada (Japan)

10:25-10:50

Break
10:50-11:05
4. Maintaining human health benefits in farmed fish: A focused review of theseafood and human health literature
11:05-11:30
Walton Dickhoff (U.S.A.)
5. NOAA/USDA Future of Feeds Initiative
11:30-11:55
Robert Iwamoto (U.S.A.)
Announcement 11:55-12:00
Lunch
12:00-13:00

Session II: Novel findings for the development of aquafeed research
6.
7.

8.

9.

(Moderators: Robert Iwamoto and Shuichi Satoh)Over view of the plant feeds in aquaculture working group13:00-13:25
Michael Rust (U.S.A.)
An overview of progress toward developing an all plant-based diet for rainbow
trout
13:25-13:50
Gibson Gaylord (U.S.A.)
Dietary supplementation strategies to improve performance of rainbow trout
Oncorhynchus mykiss fed plant-based diets
13:50-14:15
Wendy Sealey (U.S.A.)
Taurine: An essential nutrient in fish
14:15-14:40
Shin-Kwon Kim (Japan)
Break
14:40-14:55(Moderators: Walton Dickhoff and Takao Yoshimatsu)
10. Development of a new dietary material from unutilized algal resources using
fermentation skills
14:55-15:20
Motoharu Uchida (Japan)
11. Approach to expand the use of Japanese anchovy as feed material
Noriko Ishida (Japan)
15:20-15:45
12. Larval feeds research in the United States
15:45-16:10
Michael Rust (U.S.A.)
13. Aquaculture feed and seafood quality
16:10-16:35
Cheng-Sheng Lee (U.S.A.)
14. Quality control of cultured fish by feed supplements
16:35-17:00
Heisuke Nakagawa (Japan)
Announcement 17:00-17:05
2

Wednesday, October 29th, 2008

Registration

9:15-9:30

Session III: Problems and new study approaches for breakthrough(Moderators: Cheng-Sheng Lee and Makoto Yamasaki)
15. Protein production advantages in the face of increasing feed costs: Identifying
opportunities within the aquaculture industry
9:30-9:55
Gina Shamshak (U.S.A.)
16. Availability of gene-modified feed ingredient to fish
9:55-10:20
Shuichi Satoh (Japan)
17. Quantification of waste feed and fish feces using stable carbon and nitrogenisotopes
10:20-10:45
Hisashi Yokoyama (Japan)
18. Measurement of swimming behavior and digestive process of cultured fish
Toshinori Takashi (Japan)
10:45-11:10
Break
11:10-11:25
Session IV: General discussion(Moderators: Panel members)

11:25-11:55

Closing remarks (Robert Iwamoto, U.S.A. Panel Chair)

11:55-12:00

3

Presentation 1
PRESENT STATUS OF AQUAFEED RESEARCH IN JAPANTakao Yoshimatsu
Aquaculture Systems Division, National Research Institute of Aquaculture (NRIA),
Fisheries Research Agency (FRA), Mie 516-0193, Japan
E-mail: takaoyos at fra.affrc.go.jp

Japan used to be the leading fish meal manufacturer in the world. For example, in 1980s
there were constant 3-4 million mt or more of sardine Sardinops melanostictus fisheries
production from the surrounding seas of Japan. At that time this huge catch of sardine
substantially supported and maintained the large production of developing sector ofyellowtail Seriola quinqueradiata and red sea bream Pagrus major feeding cultures asmaterials for fish meal and/or Oregon-type moist pellets. However after 1990s Japanesesardine production drastically decreased mainly due to the natural regime shift ofoceanic environment with long-term intervals, and currently, we only have several
hundred thousands mt level of annual production that was scant to support the evergrowing requirements in Japanese aquaculture.
To compensate this collapse we have been importing fish meal products from
Peru, Chile and many other countries, and its total amount reaches 400-500 thousands
mt nowadays. Approximately 50% of those imported fish meal are used as a feed
ingredient for cultured fish and eventually 400-500 thousands mt of formula fish feed is
produced for finfish culture in Japan. Commercial formula feeds produced in Japan aresupplied to yellowtail culture for about 40%, to red sea bream culture for about 30%,
and to freshwater species like eel, trout and common carp for less than 20%.
For the development of aquaculture sector in Japan, NRIA played a very
important role as a governmental research institute so far. Other than the basic
nutritional and metabolic studies, recent research activities on aquafeeds in the Feed
Research Group of NRIA can be categorized as follows: 1) Efficient utilization ofplant-derived feed ingredients like soybean meal, 2) Efficient utilization of plant oil andcarbohydrate as feed ingredients for finfish, 3) Necessity of taurine as an important
nutrient for marine finfish, 4) Development of new fish feeds without fish meal using
bile salts and taurocholic acid, 5) Development of new fish feeds utilizing single-cellmaterials from marine algae, 6) Dealing with the safety issues on aquafeed ingredientsand additives, etc. Also many governmental/private universities, prefectural research
institutes and private feed companies struggle to solve problems on aquafeeds for thedevelopment of aquaculture in Japan and the world.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Yamamoto, T., Suzuki, N., Furuita, H., Sugita, T., Tanaka, N., and T. Goto. 2007.
Supplemental effect of bile salts to soybean meal-based diet on growth and feed4

utilization of rainbow trout Oncorhynchus mykiss. Fisheries Science, 73: 123-131.
A feeding experiment was conducted to evaluate the supplemental effect of bilesalts to a defatted soybean meal-based non-fish meal diet for rainbow trout
Oncorhynchus mykiss. Experimental results suggest that bile salt supplementation to asoybean meal-based diet improves the nutrient utilization by normalizing digestive
processes in rainbow trout.
2) Matsunari, H., Yamamoto T., Kim S. K., Goto T., and Takeuchi T. 2008. Optimum
dietary taurine level in casein-based diet for juvenile red sea bream Pagrus major.
Fisheries Science, 74: 347-353.
This study was conducted to investigate the effect of dietary taurine andcholyltaurine on growth and body composition of juvenile red sea bream Pagrus major.
From the experimental results it can be inferred that the optimal dietary taurine
requirement of juvenile red sea bream is 0.5% on a dry weight basis, and that the
supplementation of taurine in the diet not only improves the growth but also increases
hepatic lipid levels of red sea bream juveniles.
3) Khan M. N. D., Yoshimatsu T., Kalla A., Araki T., and S. Sakamoto. 2008.
Supplemental effect of Porphyra spheroplasts on the growth and feed utilization of
black sea bream. Fisheries Science, 74: 397-404.
A 56-day feeding trial was conducted to evaluate the effects of Porphyra
spheroplasts as a feed additive on growth, carcass composition and feed utilization of
black sea bream. The experimental results obtained in this feeding trial suggest that the
optimum dietary PS supplementation level up to 3% for black sea bream can beconsidered as improved consequence on growth performance, nutrient utilization or
body composition were noticed.

5

Presentation 2
THE ISSUES SURROUNDING AQUACULTUTE FEED AND THE CURRENT
MEASURES AGAINST THEM IN JAPANMasahide Fujise
Fish Ranching and Aquaculture Division, Fisheries Agency, Tokyo100-8907, Japan
E-mail: masahide_fujise at nm.maff.go.jp

In recent years, production of the marine fish aquaculture in Japan has remained stable
above the 250,000 t level, and occupied an important position in the Japanese marine
fishery production. However, the high price of fish meal since 2006 has kept the
inflationary price of aquaculture feed, which have afflicted fish farmers with thedeterioration of managing conditions.
Therefore, it is necessary to promote the sustainable aquaculture production,for example, through development of aquafeed that contains a little or no fish meal andthrough introduction of self-feeding systems to prevent excessive administration of
aquaculture feeds.

6

Presentation 3
CHALLENGES FOR DEVELOPMENT OF AQUAFEEDS- What should be considered for the development of commercial aquafeeds now? –Makoto Nakada
AQUARESEARCH, 2-15-11, Fujimi-shi, Saitama 354-0032 Japan,
E-mail: matona_59 at mac.com

Artificial aquafeeds which is used fishmeal as the major protein source, has been
revealed just about perfect nutrition, good-taste for raising fishes and cheap enough foraquaculture industrial use. Since world production of fishmeal and fish oil has
fluctuated dramatically further improvements in fish feed, including use of alternative
protein source, soybean meal, poultry meal and a certain amount of fish oil, along withsoybean oil, tallow and/or coconut oil, has been undertaken. We are developing theautomatic feeding systems to improve the feeding efficiency and compensate poor
nutritional value for the substitute type feeds, which are low palatability and growth.
Aquaculture in no-feeding or fertilizing system will be the new trend for world foodsupply. We should investigate the fortification feeds to those basic aquaculture types toincrease production and improve the product quality.
Nowadays, more research is needed to develop the cheap and subordinate feed,
appropriate feeding techniques that will be adopted for growth stage and seasons. Veryspecial attention should be taken to the group fish equalize feeding which are swimmingunder invisible water and difficult to care individually, refer to human andstockbreeding. Aquaculture farmer has lost profit because of raised feed prices.
Aquafeeds producers also lose benefit because of too-many kind of EP size and brandfor areas or seasons, which bring worst manufacture efficiency. Our important subject is
to generalize the unification of aquafeeds size and C/P ratio and to pay attention toreasonable feeding test plan by the fish group and the facility environment such as watertemperature and quality.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) N. Ahmed, et al, Edited by Mohammad R. Hasan, Aquaculture Management andConservation Service Fisheries and Aquaculture Management Division FAO Fisheries
and Aquaculture Department, FAO FISHERIES TECHNICAL PAPER 505, 2007,
Economics of aquaculture feeding practices in selected Asian countries, Pp1-219.
This technical paper provides an analysis of the economical implication of,and the reasons for, adopting various feeding practices for different fish species andaquaculture systems in Asia. It comprises of six selected country case study reports
from Asia (Bangladesh, China, India, the Philippines, Thailand and Viet Nam) and an
overall synthesis ending with conclusions and recommendations.
2) M. Nakada, (in press), Capture-Based Aquaculture of Yellowtail, FAO International
7

Workshop on Technical Guidelines for the Responsible use of wild fish and fisherysources for Capture-based Aquaculture production, Hanoi Viet Nam, 8-12 October 2007,
Pp 213-229.
Yellowtail mariculture has developed remarkably due to the abundant supply
and low price of wild-caught juveniles (Mojako). Ear yellowtail culture was depending
upon the locally available trash fishes such as; Sand eel, Anchovy, Mackerel, andSardine for feeds. However, it became apparent that use of trash fish as a sole feed foryellowtail, led to nutritional diseases because of putrefaction and unsuitable protein andenergy levels. High quality and raised cost of artificial feed was acceptable to yellowtail
culture, supported by expensive consumers’ price of fishes in Japan. Yellowtail
Production are stable with 60,000 tones of wild harvest and 150,000 of CBA in Japan．

8

Presentation 4
MAINTAINING HUMAN HEALTH BENEFITS IN FARMED FISH:
A FOCUSED REVIEW OF THE SEAFOOD AND HUMAN HEALTH
LITERATURE
Walton W. Dickhoff
NOAA Fisheries, Northwest Fisheries Science Center, 2725 Montlake Blvd. East,
Seattle WA 98112 USA
E-mail: Walton.w.dickhoff at noaa.gov

It is well established that the fishmeal-fish oil component of fish feeds is beneficial tofish growth and to human health for consumers of seafood. Human health benefits ofseafood include proper neurodevelopment and reducing cardiovascular disease. Animalstudies and some epidemiological studies suggest that seafood consumption may also
prevent a number of inflammatory diseases and promote mental health. A major focus
on identifying factors contributing to human health benefits has been on the long-chain
polyunsaturated fatty acids (LCPUFA) including eicosapentaenoic acid (EPA) anddocosahexaenoic acid (DHA), although other chemical components of seafood also
provide human health benefits. Since aquaculture consumes approximately 50% of the
global supply of fishmeal and 80% of the supply of fish oil, expansion of fish farming
will be limited unless effective alternatives to fishmeal and fish oil are found. Fishreared on feeds containing vegetable oils tend to have less EPA and DHA and higher
levels of other fats, including arachidonic acid (ARA), which may promoteinflammation. Responsible aquaculture should strive to maintain human health benefitsof farmed seafood by incorporating such nutritious components as LCPUFA into fish
feeds.
The strongest case for human health benefit from seafood consumption is for
reducing cardiovascular risks, including risk of heart attack and stroke. A meta-analysisof several dozen human epidemiological studies of cardiovascular disease risk indicatesthat seafood consumption to attain an intake of EPA plus DHA of 250 mg/day provides
maximal benefit. Minimal levels of LCPUFA intake to achieve health benefits forneurodevelopment and reducing risk of inflammatory disease have not been broadly
established. The appropriate ratios of EPA, DHA, and other LCPUFA to maximize
health benefit are also unknown. More studies are needed to establish the relationships
between human health parameters and seafood and LCPUFA consumption to provide
better guidelines for fish nutritionists to maintain nutritious seafood. Also, if alternatives
to fishmeal and fish oil are used, more studies of the dependence of fish growth andhealth on LCPUFA are needed.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Mozaffarian, D., and E. Rimm. 2006. Fish intake, contaminants, and human health:Evaluating the risks and benefits. JAMA 296:1885-1899.
9

The authors present a meta-analysis of human studies relating cardiovascular
events and mortality and neurological development to fish consumption, long chain n-3
fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), methyl
mercury, PCBs and dioxins. They focused on randomized clinical trials and large
prospective studies in many countries – 20 studies total. Intake of EPA + DHA of 250
mg/day decreased risk of coronary heart death by 36%. Very high rates of EPA + DHA(0.9 to 1.8 g/day as in Japan) did not further decrease risk of death, but lowered
non-fatal cardiovascular events. For neurological development, maternal DHA levels
during pregnancy positively correlated with behavioral attention, visual recognition
memory, and language comprehension of infants. With regard to methylmercury intake
with fish, the study support the advice that women of child baring age should consumemodest levels of fish but avoid four species with high mercury. Long-term exposure to
PCBs and dioxins increases cancer risk. However levels of these contaminants in fish
are equal to or lower than in beef, pork, chicken and dairy products. Higher levels of
PCBs and dioxins have been found in farmed versus wild fish, but are below levels ofconcern. Benefits of fish consumption in reduction cardiovascular death outweighcancer risk by 100- to 370-fold for farmed salmon and 300- to over 1000-fold for wild
salmon. Implications for target nutrient and contaminate levels in farmed fish can bederived from the information presented in this paper. Table 2 is a particularly usefulsummary of EPA, DHA, and contaminants in fish, shellfish and other foods.
2) Weaver, K.L., P. Ivester, J.A. Chilton, M.D. Wilson, P. Pandey, and F.H. Chilton.
2008. The content of favorable and unfavorable polyunsaturated fatty acids found incommonly eaten fish. J. Am. Diet. Assoc. 108:1178-1185.
The authors note that over the last century changes in the human diet havefavored greater consumption of saturated fat and n-6 polyunsaturated fats (PUFA) andlower n-3 PUFA. Such changes may impact cardiovascular health and promoteinflammatory diseases. With recent increases in farmed fish and changes in species offish consumed, the profile of fatty acids in the diet is changing. A survey was made of
the n-3 fatty acid composition of 30 species of the most commonly consumed farmed
and wild fish, with highest concentrations in wild and farmed salmonids and lowest
levels in tuna, red snapper, monkfish, and mahi-mahi. A more focused examination of
fatty acids was made of farmed Atlantic salmon, trout, tilapia and catfish. Farmed
salmon and trout contained higher levels (>3g/100g) of n-3 PUFA than tilapia andcatfish (<0.5 g/100g). Ratios of n-6/n-3 PUFA were well below 1 in farmed salmon andtrout, but greater than 2 in tilapia and catfish. Levels of saturated and monounsaturated
fats and arachidonic acid were also higher in tilapia and catfish. Arachidonic acid levels
were highly variable in samples depending on the source of the tilapia. Some tilapia
samples from Central America had arachidonic acid levels greater than 300 mg/110 g,
which is much higher than typically found in beef and pork. Although some laboratory
and clinical studies indicate that high ratios of n-6/n-3 PUFA promote inflammation, theideal ratio of n-6/n-3 in the human diet is controversial and may depend on genetic
factors. Results of this work raise concern about what species of fish is most appropriate
for human health benefit.
3) Fritsche, K. 2006. Fatty acids as modulators of the immune response. Annu. Rev. Nurt.
10

26:45-73.
The author provides an overview and history of the effects of n-3
polyunsaturated acids (PUFA) on such inflammation/immune-mediated human diseases
as asthma, arthritis and infection. Both laboratory and clinical studies are criticallyreviewed. Laboratory and animal studies are clearly supportive of the anti-inflammatory
nature of n-3 PUFA supplementation. Although some clinical studies clearly show somebenefit of n-3 PUFA consumption in treating inflammatory diseases, in general the
results of clinical studies are disappointing given the expectations of laboratory studies.
Some of the discordance between expectation and evidence in this area may be due toproblems with using mice as models for human disease and problems with design ofanimal and clinical trials. Recommendations are made to improve research approaches
by increasing statistical power, improving immune assays and considering genetic
variation.
4) J. Whelan and C. Rust. 2006. Innovative dietary sources of n-3 fatty acids. Annu. Rev.
Nutr. 26:75-103.
The authors briefly review scientific literature on human health benefits of n-3
fatty acids and discuss recommended levels of intake. Various potential sources ofbeneficial n-3 fatty acids are described, including vegetable oils (canola, soybean, flax),
genetically modified vegetable oils, nuts, echium oil, black currant oil, fish and fish oil,and terrestrial meats. A large summary is made of foods that have been enhanced with
n-3 fatty acids, including eggs, dairy products, margarines, salad dressings, pasta andbreads, cereal, infant formulas, baby foods, and others. Commercial sources of n-3 fatty
acids are also listed. This paper may provide a guide for fish nutritionists formulating
diets to enhance n-3 fatty acids in farmed fish.

11

Presentation 5
NOAA/USDA FUTURE OF FEEDS INITIATIVE
Robert Iwamoto* and Michael B. Rust
Northwest Fisheries Science Center, 2725 Montlake Blvd. E, Seattle, WA 98102
E-mail: Bob.Iwamoto at noaa.gov

To meet the growing consumer demand for seafood in the United States, increasing
supplies of finfish and shellfish will be needed. Most experts agree that development ofaquaculture will be the only way to sustainably meet this increase in demand. The
question that must be answered is how to ensure that aquaculture production increases
are sustainable. The development and expansion of farming of carnivorous fish species
will be constrained by a limited supply of fish meal and oil for feeds. Fishmeal and oil
have traditionally made up a large part of the diet of farm-raised carnivorous fish.
Fortunately, there is no dietary requirement for specific amounts of fish meal or fish oil
for fish, so feeds that lessen the reliance on these limited feedstuffs—such as alternative
protein and oil resources—can, and must, be developed. For this reason, the U.S.
Department of Commerce (NOAA) and the U.S. Department of Agriculture (USDA) are
jointly sponsoring an aquaculture feeds initiative to address those issues. The initiative
primarily consists of expert and public consultations that will discuss the future of fish
feeds and the benefits to the U.S. by the development of such alternative feeds.
Elements of the initiative include:
1. An invitation to the general public to comment on the issue, which will indicate
the level of understanding and knowledge that the public has regarding fish
feeds.
2. A consultation with experts who are active researchers in the area of fish feeds
and related topics (Researcher Panel).
3. A consultation with experts who are active stakeholders in the area of fish feeds
and related topics (Stakeholder Panel).
4. A White Paper answering the questions raised by the public comment process
and summarizing the results of the two expert panels.
5. A public meeting to summarize the contents of the White Paper and to allow forcomment by the broader stakeholder community.
A Steering Committee made up of scientists, federal policymakers, communications,public relations experts and logistics experts has been assembled to move the process
through these five steps.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) NOAA/USDA ALTERNATIVE FEEDS INITIATIVE Robert Iwamoto and Michael
12

Rust, Northwest Fisheries Science Center, 2725 Montlake Blvd E, Seattle, WA, 98102
USA. Alternative Feeds Initiative Web Site.
http://aquaculture.noaa.gov/news/feeds.html
The National Oceanic and Atmospheric Administration (NOAA) in partnershipwith the U.S. Department of Agriculture (USDA) launched the Alternative Feeds
Initiative on November 15, 2007, to accelerate the development of alternative feeds foraquaculture. Details of meetings and public comments can be found at this site. This
web site track it’s progress and will be used to provide copies of the white paper when it
is finished2) NOAA 10-Year Plan for Marine Aquaculture, October 2007. Available at
http://aquaculture.noaa.gov
Takes input from reports by the Presidents Commission on Ocean Policy, theNational Marine Aquaculture Summit and stakeholder input to develop a strategy andtactical plan for the NOAA aquaculture program.
3) Offshore Aquaculture in the United States: Economic Considerations, Implications &Opportunities, July 2008 (Pre-Publication Copy).
Available at http://aquaculture.noaa.gov
Ten authors cover 12 chapters on various aspects that impact the economic
considerations that may apply to offshore aquaculture. Chapters cover: US potential,
feeds, technologies, analogous industries, economic models, national impacts,
interactions with wild capture fisheries, current status, markets and the issues
surrounding the current debate in the US.
4) Summary of the National Marine Aquaculture Summit. June 2007. Available at
http://aquaculture.noaa.gov
Summary of a summit of aquaculture leaders, health professionals and seafood
stakeholders addressing the needs for and ramifications of developing a significant
marine aquaculture sector in the United States.
5) US Department of Commerce Aquaculture Policy. Available at
http://aquaculture.noaa.gov
Provides the policy framework for aquaculture research, development andregulation for NOAA and other agencies of the Department of Commerce.
6) Presendents Ocean Commission, 2004. Setting a course for marine aquaculture –Chapter 22. An ocean blueprint for the 21st century. Pages 330-336. Available at
http://aquaculture.noaa.gov
Report of a special commission on the Oceans to recommend policy for the
President of the United States. Chapter 22 deals with marine aquaculture and provides
much of the rational for an invigorated US effort in marine aquaculture.

13

Presentation 6
OVER VIEW OF THE PLANT FEEDS IN AQUACULTURE WORKINGGROUP
Michael B. Rust
Aquaculture Program, Northwest Fisheries Science Center, 2725 Montlake Blvd E,
Seattle, WA, 98102 USA
E-mail: mike.rust at noaa.gov

A large number of researchers from several countries and representatives from US
federal technical agencies came together on several occasions to develop an integrated(cross-disciplinary), strategic approach to improve understanding of the factors limiting
inclusion of plant feedstuffs in the diet of carnivorous fishes, with the goal to increase
inclusion of sustainable plant nutrients in feeds for aquaculture. A roadmap of theresearch required to reach that goal was drafted. The specific objectives of the working
group were to define goals, performance measures, present status, and two and four year
targets. The defined goals are; 1) Establish standardized research approaches andprotocols for systematic evaluation of plant feedstuffs across carnivorous fish species.
2) Enhance fish germplasm and discover genes. 3) Enhance the inherent composition ofcrops to provide a beneficial balance of bioactive compounds in order to optimize theiruse in aquafeeds for carnivorous fish. 4) Improve understanding of the interactions
between plant-based diets and intestinal microflora and their effects on the intestinal
morphology, nutrient transport, gut-associated immune responses, and disease resistancein carnivorous fish species. 5) Improve and optimize ingredient processing, feed
manufacturing technology, and feed formulations to increase inclusion of plant-derived
ingredients in the diet of carnivorous fish. 6) Optimize the storage, nutritional andsensory quality of aquaculture species for human consumption. 7) Establish aninternational communications network for research on optimizing plant products inaquafeeds. This information has been written into a strategic plan which outlines
research needs. In addition two reviews on the use of plant feeds in aquaculture havebeen produced by the working group, and a tactical plan is being developed. A web site(http://www.aquafeed.com/ppa-about.php) has been established to encouragecoordination and international collaboration to improve the use of plant products inaquafeeds for species grown in all parts of the world. Participation is open to all
researchers interested in this field. Those interested should visit the web site.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Over view of the plant feeds in aquaculture working group. Michael B. Rust,
Aquaculture Program, Northwest Fisheries Science Center, 2725 Montlake Blvd E,
Seattle, WA, 98102 USA. Plant Products in Aquafeeds Working group. 2005.
Expanding the Utilization of Sustainable Plant Products in Aquafeeds White Paper.
Available at http://www.aquafeed.com/ppa-about.php
14This “white paper” considers various types of plant feedstuffs which currently
are or potentially may be incorporated into aquafeeds to support the sustainableproduction of various fish species in aquaculture. The nutritional composition of thesevarious feedstuffs are considered along with the presence of any bioactive compounds
that may positively or negatively affect the target organism. Specific strategies andtechniques to optimize the nutritional composition of plant feedstuffs and limit
potentially adverse effects of bioactive compounds are also described.
2) Plant Products in Aquafeeds Working group. 2005. Aquafeed Working Group
Strategic Research Plan available at
http://www.aquafeed.com/ppa-about.php
This paper provides a guide for coordinating and enhancing the development of
new research efforts. It is a comprehensive and coherent research agenda for addressing
the problems encountered when increasing the amount of plant protein used in feed for
carnivorous fish species. Seven writing teams drafted a “roadmap” of the research steps
or performance measures needed to accomplish the seven goals identified by the group.
With a plan that will stand up to review by peers and program managers, researchers
have the competitive advantage when responding to solicitations for proposals, andwhen proposing new programs.
3) Gatlin, Delbert, M. III, Frederic T Barrows, Paul Brown, Konrad Dabrowski, T
Gibson Gaylord, Ronald W Hardy, Eliot Herman, Gongshe Hu, Åshild Krogdahl,
Richard Nelson, Kenneth Overturf, Michael Rust, Wendy Sealey, Denise Skonberg,
Edward J Souza, David Stone, Rich Wilson, Eve Wurtele. 2007. Expanding theutilization of sustainable plant products in aquafeeds: a review Aquaculture Research,
Volume 38, Issue 6: 551-579
Based on the 2005 white paper “Expanding the Utilization of Sustainable Plant
Products in Aquafeeds” this peer-reviewed publications expands on the original.
4) Barrows, Frederic T., Diane Bellis, Åshild Krogdahl, Jeffrey T. Silverstein, Eliot M.
Herman, Wendy M. Sealey, Michael B. Rust and Delbert M. Gatlin III. 2008. Report of
the Plant Products in Aquafeed Strategic Planning Workshop: An Integrated,
Interdisciplinary Research Roadmap for Increasing Utilization of Plant Feedstuffs inDiets for Carnivorous Fish. Reviews in Fisheries Science, Volume 16, Issue 4:449 – 455.
Based on the Plant Products in Aquafeeds Strategic plan, this peer-reviewed
publication expands on the original.

15

Presentation 7
AN OVERVIEW OF PROGRESS TOWARD DEVELOPING AN ALL
PLANT-BASED DIET FOR RAINBOW TROUT
T. Gibson Gaylord1*, Frederic T. Barrows1, Kenneth E. Overturf1, Keshun Liu2, andGongshe Hu2
1

USDA-ARS Small Grains and Potato Germplasm Research Unit, Hagerman, Idaho,
USA
2
USDA-ARS Small Grains and Potato Germplasm Research Unit, Aberdeen, Idaho,USAE-mail: Gibson.Gaylord at ars.usda.gov

Feeds for rainbow trout have historically contained high quantities of fish meal and fish
oil as cost effective sources of both essential amino acids and fatty acids. Research
results from around the world have identified ingredients to replace fish meal and fishoils, but limits still exist in completely removing these ingredients from the diet.
The USDA-ARS Trout-Grains Project, officially titled “The integration ofnutritional, genetic and physiological approaches to improve production efficiency ofrainbow trout (Oncorhynchus mykiss)” was initiated with a primary goal to identify costeffective replacements for fish meal. The project team consists of two fish nutritionist, afish geneticist, a cereal grain chemist and a plant geneticist. This group of scientist
addresses a range of issues including; selection/development of cereal grains withimproved nutritional profiles, development of methods to fractionate valuable
components of the grains, and development of families of trout that are better able toutilize nutrients from plant-based feeds. The nutrition component of the project focuseson evaluation of new and traditional ingredients, nutrient requirements for specific life
stages, and identification of nutrients in fish meal not present in plant meals.
Finally we are developing a database of nutrient availability from plant-based
ingredients that will have potential to supply the nutritional needs of rainbow trout inaquaculture.
Some highlights of our research findings have been:
1.) Taurine may be conditionally indispensable when only plant-derived protein
sources are utilized.
2). Vitamin premix formulations require modification when plant derived ingredientsare utilized in lieu of fish meal in extruded trout feeds.
3) Starch amylose/amylopectin ratios in energy feed ingredients will affectcarbohydrate and energy digestibility.
Results of laboratory and pilot scale studies of feeding fish meal free diets torainbow trout will be presented.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) USDA-ARS Trout-Grains Project. Developing Plant-Based Feeds for Rainbow Trout.
16

Barrows, F., Gaylord, T.G., Sealey, W.M., Smith, C.E., Porter, L. 2008. The effect ofprotein source and vitamin premix on growth efficiency of rainbow trout, Oncorhynchus
mykiss. Aquaculture 283:148-155.
The authors tested the adequacy of current NRC guidelines for minimal vitamininclusion levels in fishmeal-based and plant-based diets for rainbow trout. The authors
estimated losses of vitamins due to cooking extrusion processing of the feeds anddetermined that vitamin premixes required adjustments based in dietary ingredients(fishmeal vs. plant-based). Fish performance variables showed an interaction between
vitamin inclusion levels and plant vs. fish based diets. The authors therefore
recommended a modified vitamin premix formulation that adjusted for processing andstorage loss in plant-based trout feeds.
2) Lunger, A.N., Mclean, E., Gaylord, T.G., Kuhn, D., Craig, S.R. 2007. Taurine
supplementation to alternative dietary proteins used in fish meal replacement enhances
growth of juvenile cobia (Rachycentron canadum). Aquaculture. 271:401-410
The authors found that fish meal levels could be reduced to less than 15% of thediet for juvenile cobia when amino acids and taurine were supplemented with increasinglevels of yeast protein. Taurine supplementation appeared to be a major factor incontributing to the yeast protein replacement in the authors’ first trial. The authors’
second trial showed less positive results, but may have been due to other limiting aminoacids inhibiting performance. The authors highlight the importance of quantitative
amino acid requirements for cobia and the potential improvements in growth and feed
efficiency that taurine supplementation may have in alternative ingredient diets fed tocobia.
3) Gaylord, T.G., Barrows, F., Teague, A.M., Johansen, K.A., Overturf, K.E., Shepherd,
B.S. 2007. Supplementation of taurine and methionine to all-plant protein diets forrainbow trout (Oncorhynchus mykiss). Aquaculture 269:514-525.
The authors tested the ability to supplement methionine and/or taurine toplant-based diets fro rainbow trout. The authors observed a small improvement inweight gain with taurine supplementation while methionine supplementation inhibited
growth. Taurine supplementation also improve fillet yield in trout when no methionine
was supplemented. The authors discuss the confounding responses of methionine andtaurine supplementation in plant-based trout feeds and the need for further research toaddress optimal supplementation levels of amino acids.
4) Gaylord, T.G., Teague, A.M., Barrows, F.T. 2006. Taurine supplementation of
all-plant protein diets for rainbow trout, Oncorhynchus mykiss. Journal of the World
Aquaculture Society. 37:509-517.
The authors address the potential for taurine supplementation in all-plant protein
based diets for rainbow trout in a factorial design experiment utilizing diets with andwithout fishmeal protein. Taurine supplementation improved growth, feed conversion
ratios, protein retention efficiencies and energy retention efficiencies of fish fed theplant protein diets. No effects of taurine supplementation were observed for these
response factors in fish fed the diets containing fishmeal. The authors concluded thattaurine is conditionally indispensable to trout fed plant-based feeds.
175) Gaylord, T.G., Barrows, F.T., Rawles, S.D., Liu, K., Bregitzer, P., Hang, A., Obert, D.,
and Morris, C. (2008) Apparent digestibility of nutrients in extruded diets from cultivarsof barley and wheat selected for nutritional quality in rainbow trout (Oncorhynchus
mykiss). Aquaculture Nutrition. On-line early
The authors addressed the potential differences in feed value of lines of barley
and wheat selected for differences in nutrient composition. The authors performed adigestibility experiment and found that phosphorus availability ranged from 17 to 78%
and was influenced by starch type in wheat. Apparent protein digestibility ranged from53 to 125% and differences were observed between wheat varieties based on starch type.
Apparent energy digestibility ranged from 32 to 63%, with waxy barley varieties having
higher energy digestibility coefficients than normal starch varieties. Waxy starchvarieties had higher starch digestibility in both barley and wheat because of the greater
digestibility of amylopectin than amylose. The authors concluded that the higher energy
digestibility of waxy barley lines suggests that these varieties warrant further attention
as feed ingredients for rainbow trout.
6) Barrows, F., Gaylord, T.G., Sealey, W.M., Haas, M.J., Stroup, R.L. 2008. Processing
soybean meal for biodiesel production; effect of a new processing method on growth
performance of rainbow trout, Oncorhynchus mykiss. Aquaculture.283:143-147.
A new method of soybean meal processing has been developed, which may
simplify the process of biodiesel production. This method, ‘in situ transesterification’,eliminates hexane extraction to remove the oil, combining the extraction andtransesterification steps so as to synthesize biodiesel via a single treatment conducted
directly on a lipid-bearing solid material. If the resulting meal is comparable innutritional value to commercially available hexane extracted soybean meal (SE-SBM)
the new process could become widely used in the bio-fuel industry. Two levels (17.5
and 35%) of each of three types of soybean meal were fed to triplicate lots of rainbow
trout. The three types of soybean meal included SE-SBM, experimentally produced
hexane extracted SE-SBM (ESE-SBM), and meal produced using in situ
transesterfication (IS-SBM). There was no effect of source of soybean meal on weightgain of trout. The fish fed the meal processed by the new method, IS-SBM, gained asmuch weight as fish fed either of the two control meals, within an inclusion level. The
fish fed the diets containing IS-SBM, however, did have higher feed intakes (2.51%bw/d) as compared to fish fed the ESE-SBM or SE-SBM, 2.38 and 2.46% bw/d,
respectively. Protein and energy retention values were lower for the fish fed the IS-SBM
diets. ADC for protein was lower for the IS-SBM (85.9%) than for the ESE-SBM(89.3%). Feeding IS-SBM did not decrease weight gain in this study, but due to theincreased feed intake and FCR, long term feeding trials should be conducted to further
evaluate the meal.

18

Presentation 8
DIETARY SUPPLEMENTATION STRATEGIES TO IMPROVEPERFORMANCE OF RAINBOW TROUT Oncorhynchus mykiss FED
PLANT-BASED DIETS
Wendy M. Sealey1*, Frederic T. Barrows2, Charlie E. Smith3 and Ronald W. Hardy1
1

Hagerman Fish Culture Experiment Station, University of Idaho, 3059F National Fish
Hatchery Road, Hagerman, ID 83332, USA
2
Trout Grains Program, United States, Department of Agriculture, Agricultural ResearchService, 3059F National Fish Hatchery Road, Hagerman, ID 83332, USA
3
Aquaculture & Pathology Consulting, Bozeman, MT 59715, USA
E-mail: wsealey at uidaho.edu

Higher inclusion levels of plant-based feedstuffs in diets for rainbow trout necessitate
identification of methods to mitigate the negative effects on fish growth and health.
Thus, an increasing number of novel and re-purposed dietary supplements are becomingcommercially available to counteract the anti-nutritional effects of plant-based
ingredients. Specifically, probiotic bacteria and yeast products, which have previously
been primarily investigated for their anti-pathogenic effects in fish, are now promoted toaid in plant-based diet utilization and digestive tract health. Additionally, dietary
supplementation of commercially produced anti-inflammatory antibodies used to reduce
gastrointestinal tract inflammation in terrestrial animals may also have potential to
improve digestive tract health and performance of aquatic animals fed plant-based diets.
The present paper describes a series of studies conducted at the Hagerman Fish Culture
Experiment Station that investigated the ability of dietary probiotics, yeast, andanti-phospholipase A2 antibody to improve performance of rainbow trout fed soybean
meal-based diets.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Bakke-McKellep, A.M., M.H. Penn, P.M. Salas, S. Refstie, S.Sperstad, T. Landsverk,
E. Ringo and A. Krogdahl. 2007. Effects of dietary soybean meal, inulin andoxytetracycline on intestinal microbiota and epithelial cell stress, apoptosis andproliferation in the telost Atlantic salmon (Salmo salar L.) British Journal of Nutrition,
Volume 97. Pp 699-713.
The authors explore the role of the intestinal microbiota of salmon in thepathogenesis of soybean meal-induced enteritis. A comprehensive approach is described
involving identification of both descriptive and functional indices of gut health.
Detailed methodology for the varied indices is provided allowing for more thoroughevaluation of the effects of alternative protein sources in salmonids as well as lessstudied species. Results of the study indicate that soybean meal-induced enteritis isaccompanied by induction of distal intestine epithelial cell responses and changes in19

microbiota. These results strengthen the putative linkage between gut health andmicrobiota and indicate that further research in this area is warranted.
2) Gatesoupe, F.J. 2007. Live yeasts in the gut: Natural occurrence, dietary introduction
and their effects on fish health and development. Aquaculture. Volume 267. Pp 20-30.
The author summarizes the present state of knowledge regarding the importanceof yeast in the fish gut. The putative role of yeast is discussed in regards to both thenatural occurrence and the use of “industrial” yeast in aquaculture. Functions discussed
include pathogencity, colonization and competition, modulation of immune responses
and alteration of host (fish) metabolism. Species, and yeast subcomponents ofimportance (i.e. glucans) are identified and supporting references are included.
3) Kesarcodi-Watson, A., H. Kaspar, M. J. Lategan, and L. Gibson. 2008. Probiotics inaquaculture: The need, principles and mechanisms of action and screening processes.
Aquaculture. Volume 274. Pp 1-14.
The authors review recent and historic literature regarding the use of probiotics
in aquaculture. Special emphasis is placed on differences between the use of probiotics
in mammalian and aquatic species. These differences illustrate and the authors
described why the definition of probiotic as described by Fuller (1989), “a live
microbial feed supplement which beneficially affects the host animal by improving its
intestinal balance” must be expanded for aquatic species.
Definitions which more accurately reflect the mode of actions observed forprobiotics in aquatic species are proposed. Modes of action including improving the useof feed or enhancing the nutritional value are discussed.

20

Presentation 9
TAURINE: AN ESSENTIAL NUTRIENT IN FISH
Shin-Kwon Kim 1*, Masahito Yokoyama 1, Toshio Takeuchi 2
1

Aquaculture Biology Division, National Research Institute of Aquaculture, FRA, Mie
516-0193, Japan
2
Tokyo University of Marine Science and Technology, Tokyo 108-8477, Japan
E-mail: ksk4116 at affrc.go.jp

Fish meal is the most important protein source in aquaculture feed for finfish. In
addition to protein, fish meal contains many functional substances such as essential fatty
acids, vitamins, minerals, and so on. Fish meal also contains much amount of taurine,
which is recently considered to be a candidate nutrient having potential as a growthpromoter for marine finfish. We conducted some dietary experiments by feeding taurine
free diet for Japanese flounder to elucidate the nutritional essentiality of taurine in fish.
Different taurine level diets were prepared by the supplementation of taurine tothe basal composition. Taurine was eliminated from fish meal as low as possible bywashing with 70% ethanol. This fish meal was used as the sole protein source forexperimental diets.
Abnormal feeding behavior was observed in the taurine deficient fish group,
whereas normal in the taurine supplemented group, during experimental period. Fish fed
taurine deficient diet showed low growth performance. The taurine contents of thewhole body and tissues increased with increase of the taurine supplementation. The
concentrations of taurine conjugated bile acids, such as taurocholic acid andtaurochenodeoxycholic acid, in the gall bladder increased by feeding the high taurine
level diet. These findings of the taurine deficiency indicate that taurine is essential
nutrient for normal growth and development of Japanese flounder.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Kim, S.K., Matsunari, H., Takeuchi, T., Yokoyama, M., Furuita, H., Murata, Y., Goto,
T. 2008. Comparison of taurine biosynthesis ability between juvenile of Japaneseflounder and common carp. AMINO ACIDS, Vol:35:1. Pp 161-168.
This study was conducted to investigate taurine deficiency and the ability of
taurine biosynthesis in both juvenile Japanese flounder (JF) and juvenile common carp
(CC) in vivo using low taurine level diets. Three different taurine level diets wereprepared by the supplementation of taurine to the basal composition (JF – 0, 0.5 and1.5% in JF; CC – 0, 1, 3% in CC). The final average body weight and feed efficiency of
JF fed the JF – 1.5% was significantly higher than those of fish fed on the JF – 0%. On
the other hand, no significant difference was observed in CC fed with CC – 0, 1, and
3% diets. The taurine retention rate was negative in the case of JF-fed with thetaurine-free supplement (JF – 0%). On the other hand, the taurine retention rate wasabout 280% in the case of CC-fed with the taurine-free supplement (CC – 0%). These21

findings indicate that while taurine is essential for growth of JF, it is not essential for thegrowth of CC.
2) Kim, S.K., Matsunari, H., Nomura, K., Tanaka, H., Yokoyama, M., Murata, Y.,
Ishihara, K., Takeuchi, T. 2008. Correlation of dietary taurine levels and lipid contents
in diets on the conjugated bile acid composition and growth performance of juvenile
Japanese flounder Paralichthys olivaceus. Fisheries Science, Vol:74:4. Pp 875-881.
The effects of dietary taurine levels and lipid contents on the conjugated bile
acid composition and growth performance of juvenile Japanese flounder Paralichthys
olivaceus were investigated. Six types of diet (three different levels of taurine at two
different levels of lipid) were fed to juveniles (average body weight, 0.04 g). Fishmealthat was washed with 70% ethanol to remove taurine was used as the sole protein source.
Feeding experiments were carried out at 20°C for 6 weeks. The body weight andpercent weight gain of the juveniles were improved by the dietary taurine
supplementation. The taurine contents of the whole body and tissues increased with theincrease of the dietary taurine contents. The conjugated bile acids in the gall bladderconsisted of taurocholic acid and taurochenodeoxycholic acid, which increased with theincrease of the dietary taurine level. Taurocholic acid accounted for more than 95% of
the total conjugated bile acids. This indicates that taurine is the sole amino acid toconjugate bile acid in Japanese flounder.
3) Kim, S.K., Matsunari, H., Takeuchi, T., Yokoyama, M., Murata, Y., Ishihara, K. 2007.
Effect of different dietary taurine levels on the conjugated bile acid composition andgrowth performance of juvenile and fingerling Japanese flounder Paralichthys olivaceus,
Aquaculture, Vol:273. Pp 595-601
This study investigated the effects of different taurine levels on the conjugatedbile acid composition and growth performance of juvenile and fingerling Japaneseflounder. Five level diets of dietary taurine were prepared by the supplementation oftaurine (0, 0.1, 0.2, 0.5 and 1.5%) to a basal composition. Fish meal washed with 70%
ethanol to remove taurine was used as the sole protein source. Two feeding experiments
were carried out at 20 °C by using different sized fish (BW: 0.7 g and 9.6 g). Both sizegroups of fish were fed the experimental diets for 6 weeks. The body weight and feed
efficiency of Japanese flounder were improved by taurine supplementation in theexperimental diets. The taurine contents of the whole body and tissues increased with
the increase of the taurine supplementation. The conjugated bile acids in gall bladderwere composed with taurocholic acid (Expt.I: 33.0–146.8 mg/ml; Expt.II: 64.8–145.3
mg/ml) and taurochenodeoxycholic acid (Expt.I: 0.8–5.5 mg/ml; Expt.II: 3.5–4.9
mg/ml). These bile acids increased with the increase of the dietary taurine level.
Taurocholic acid accounted for more than 95% of the total conjugated bile acids. No
other conjugated bile acids except the taurine conjugated bile acids were detected in thegall bladder of Japanese flounder. This means that taurine is the sole amino acid toconjugate bile acid in Japanese flounder. Dietary taurine intake affects the conjugatedbile acid composition in juvenile and fingerling Japanese flounder.
4) Kim, S.K., Takeuchi, T., Yokoyama, M., Murata, Y., Kaneniwa, M., Sakakura, Y.
2005. Effect of dietary taurine levels on growth and feeding behavior of juvenile
22

Japanese flounder Paralichthys olivaceus. Aquaculture, Vol:250. Pp 765-774.
This study was conducted to investigate the effect of dietary taurine levels on
growth and feeding behavior of juvenile Japanese flounder. Three different taurine level
diets were prepared by supplementation of taurine (T-0%, 0.5% and 1.5%) to the basal
diet. Fish meal washed with 70% ethanol to remove taurine was used as the sole protein
source. Feeding experiments were carried out twice at 20°C by using different size offish (average body weight: 0.3 g in Experiment I and 3.7 g in Experiment II). The
feeding behavior of fish was observed throughout the experimental period. The final
average body weight and feed efficiency of juvenile Japanese flounder fed the T-1.5%
diet was significantly higher than those of fish fed the T-0% diet in Experiments I and II.
Abnormal feeding behavior such as multiple feeding while swimming in the watercolumn was observed in the T-0% group in Experiment I. These findings indicate thattaurine is essential for normal growth and development of normal feeding behavior ofjuvenile Japanese flounder.

23

Presentation 10
DEVELOPMENT OF A NEW DIETARY MATERIAL FROM UNUTILIZEDALGAL RESOURCES USING FERMENTATION SKILLS
Motoharu Uchida*, Tatsuo Miyoshi
National Research Institute of Fisheries and Environment of Inland Sea, FRA,
Hiroshima 739-0452, Japan
E-mail: uchida at affrc.go.jp“Marine silage (MS)” is fish dietary materials prepared from algae by a fermentative
processing. On the 34th U.S.-Japan Aquaculture Panel Symposium, we had reported themethod to prepare the MS from seaweed by lactic acid fermentation. The objective of
this work is to report the recent progress of this study.
The fermentation of seaweeds can be performed by the enzymaticsaccharification by cellulase, followed by the fermentation process with the use of lactic
acid bacteria. This method can be applied on any kind of seaweeds, but quantity oflactic acid production was varied at least more than six times among the different lot
samples of seaweed. Lactic acid fermentation of microalgae is also possible, but
decomposition of cell wall is only partial with presently available enzyme products.
Developing MS is interesting from three viewpoints. Firstly, it makes possible
to convert unutilized algal biomass resources into dietary materials for aquaculture.
Secondly, fermentation product is acidic and long-preservative at room temperature
without energy cost. Therefore, for example, surplus microalgae culture can bepreserved and supplied depending on a demand. Thirdly, fermented materials areusually expected to have some functions to contribute to keep health. Red sea breamthat are challenged with iridovirus and fed a diet containing the MS at 10% (w/w)
resulted in the significant promotion of the survival rate (%) against those fed thecontrol diet. We believe that aquaculture with MS is eco-friendly and contribute toculture fish that is free from drugs.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Uchida, M., and M. Murata. 2002. Fermentative preparation of single cell detritus
from seaweed, Undaria pinnatifida, suitable as a replacement hatchery diet forunicellular algae. Aquaculture, 207. 345-357.
Single cell detritus (SCD) is a product ca. 5.8-11.5μm in diameter prepared bydecomposing seaweed to a cellular level, and is expected to be utilized as afisheries–diet in place of unicellular algae. The present paper details the culture
conditions for the effective preparation of SCD from Undaria pinnatifida based on afermentative method. The presently developed method needs no facilities fortemperature conditioning, lightening, and air supplying, but needs only a plastic tank for
obtaining the SCD. Here, we suggest a novel fisheries-diet of “marine silage”, which is
produced from the combination of two principles, conversion of seaweed to SCD and24

induction of lactic acid fermentation utilizing activities of a microbial mixture
composed of a lactic acid bacterium and yeast.
2) Uchida, M., and M. Murata. 2004. Isolation of a lactic acid bacterium and yeast
consortium from a fermented material of Ulva spp. (Chlorophyta). Journal of Applied
Microbiology, 97. 1297-1310.
Three kinds of microorganisms composed of a lactic acid bacterium,
Lactobacillus brevis, and yeasts, Debaryomyces hansenii and a Candida
zeylanoides-related specimen, were isolated from a fermented material of Ulva spp.
Lactic acid and ethanol fermentation could be induced in various kinds of seaweed byadding this microbial consortium along with cellulase. This is the first report ofdemonstrative performing of lactic acid and ethanol fermentation in seaweed, which is
expected to provide a new material for food and dietary applications.
3) Uchida, M., K. Numaguchi, and M. Murata. 2004. Mass preparation of marine silage
from Undaria pinnatifida and its dietary effect for young pearl oyster. Fisheries Science,
70. 456-462.
Marine silage (MS) is a new dietary item prepared by decomposing seaweed toa cellular unit and performing lactic acid fermentation. The present paper demonstrated
a large (10 L)-scale preparation and a long-term (18 months)-preservation of the MS
using Undaria pinnatifida as a substrate. Furthermore, feeding trials demonstrate the
dietary value of the MS to the Japanese pearl oyster Pinctada fucata martensii. The
present study is the first to demonstrate a mass preparation of MS and its positive
dietary effect for a bivalve.
4) Uchida, M., H. Amakasu, Y. Satoh, and M. Murata. 2004. Combinations of lactic acidbacteria and yeast suitable for preparation of marine silage. Fisheries Science, 70.
508-518.
To examine a suitable combination of starter microorganisms, marine silage(MS) was prepared with different compositions of LAB and yeasts from a substrate of
Undaria pinnatifida. Lactobacillus brevis showed the highest predominating ability in
MS among the tested, while L. acidophilus (casei-type) IAM 10074 and L. plantarum
IAM12477T also showed predominating ability with relatively inferior scores. The
present study demonstrated that a single use of LAB, including food industry-familiar
species, is suitable for preparation of MS, while the superiority of L. brevis B5201 wassuggested in predominating ability in marine silage.

25

Presentation 11
APPROACH TO EXPAND THE USE OF JAPANESE ANCHOVY AS FEED
MATERIAL - Efficacy of thiamine supplementation Noriko Ishida1*, Kenji Nakanishi2, and Hisashi Murata3
1

National Research Institute of Fisheries Science, FRA, Yokohama 236-8648, Japan
Miyazaki Prefectural Fisheries Experimental Station, Miyazaki 889-2162, Japan
3
University of Miyazaki, Miyazaki 889-2192, Japan
E-mail: larry at affrc.go.jp
2

Japanese anchovy (Engraulis japonicus) resource is considered to be extensive,
however not effectively utilized. In order to resolve this issue and eventually increase
the anchovy catch, Fisheries Agency has launched a project study for effective
utilization of anchovy to increase the value of anchovy.
Here, we introduce the project study schema and five research issues
respectively, such as fish quality, fish machinery, surimi, fish meal, and feed material.
Then, we would like to focus on the results of the feed material issue.
Moist pellets are still used as a major feed material in amberjack (Seriola
dumerili) farms in western Japan. Among the fish species used for feed materials,
anchovy is definitely the cheapest. However anchovy is not often applied to moist
pellets for farming amberjack because of the development of thiamine deficiency. It issaid that thiamine deficiency developed in farming yellowtail (Seriola quinqueradiata)
which were fed anchovy. The amberjack farmers are concerned that the same diseasedevelopment will occur in amberjack and do not use anchovy for moist pellets.
From these circumstances, we investigated the development of thiamine
deficiency in farmed amberjack. We designed three experimental groups which wererespectively fed anchovy-based moist pellets, thiamine supplemented anchovy-based
moist pellets, and mackerel-based moist pellets that the farmers often use. These diets
were adjusted to the same caloric value. We concluded that thiamine supplementation isnecessary for anchovy-based moist pellets. The thiamine supplementation for anchovypellets prevented thiamine deficiency disease, but also achieved as the same level of thegrowth, disease-resistance, and meat quality as the cultured fish which was fed
mackerel-based moist pellets.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Ishihara, T., M. Yasuda, and S. Kashiwagi. 1974. Studies on thiaminase I in marine
fish- VI Preventive effect of thiamine on nutritional disease. Bull. Japan. Soc. Sci. Fish.,40 (8): 775-781.
Yellowtail fed on raw anchovy as a sole feeding stuff with or without thiamine
was analyzed for thiamine, pyruvic acid, lipid and protein contents in various tissues.
The results obtained may be summarized as follows: (1) In the fish kept on raw anchovywith no added thiamine, contents of thiamine, cholesterol and triglyceride in various
26tissues were significantly decreased, while pyruvic acid in the blood increased. The
pyruvic acid and thiamine contents however, were restored to normal levels whenthiamine was added to the meat. (2) The fish reared on raw anchovy with added
thiamine during the entire period showed a similar body composition to the fish given
horse mackerel, mackerel and sand launce which contain no thiaminase. [In Japanesewith English summary]2) Masumoto, T., R. W. Hardy, and E. Casillas. 1987. Comparison of transketolase
activity and thiamin pyrophosphate levels in erythrocytes and liver of rainbow trout(Salmo gairdneri) as indicators of thiamin status. American Inst. Nutr., 117: 1422-1426.
Yearling rainbow trout (Salmo gairdneri) were fed a purified diet with andwithout thiamin supplementation for 30 wk, at which time overt signs of thiamindeficiency appeared in the deficient group. Overt signs of thiamin deficiency wereanorexia, darkening and ataxia. Death rapidly followed the development of overt
thiamin deficiency. Transketolase activity and thiamin pyrophosphate levels were
measured monthly in erythrocyte and liver samples. Significant differences inerythrocyte transketolase activity between fish fed the thiamin-deficient and control
diets were measured after 24 wk of feeding. No significant difference in livertransketolase activity was found between rout fed diets with or without thiaminsupplementation. Thiamin pyrophosphate levels were significantly lower in erythrocytes
and liver of fish fed the thiamin-deficient diet after 16wk of feeding. Thiamin
pyrophosphate levels in erythrocytes and liver were found to be a more sensitive
indicator of thiamin status of rainbow trout than erythrocyte or liver transketolase
activity.
3) Nyoman Adiasmara Giri, I., A. Kanazawa, S. Teshima, and S. Koshio. 1996. Effects
of dietary thiamin on growth and thiamin content on the hepatopancreas of kuruma
prawn (Penaeus japonicus) juveniles. Suisanzoshoku, 44 (3): 325-333.
Penaeus japonicus (0.99 ± 0.06g) were fed with diets containing 0, 5, 15 and 20
mg thiamin per 100g dry diet for 12 weeks to investigate their effects on survival,
growth and thiamin content in the hepatopancreas of prawns. Prawns fed
thiamin-deficient diet showed significantly lower survival, weight gain, and thiamin
pyrophosphate (TPP) content in the hepatopancreas than those from other treatments.
While a significant difference in weight gain was first observed by the eighth week offeeding, the TPP content in the hepatopancreas of prawn fed thiamin-deficient diet wasmarkedly decreased already by the fourth week of feeding, being also significantly
lower compared to other treatments at the end of experiment. These results indicatedthat the TPP content in the hepatopancreas may be more sensitive indicator to detect thehealthy condition of prawn than growth. A positive relationship between weight gain
and TPP content in the hepatopancreas was found in the present study. Based on theweight gain and on TPP content in the hepatopancreas of prawn, the minimum thiaminrequirements of kuruma prawn juveniles were estimated to be 13.98 mg and 14.31 mg
per 100g dry diet, respectively.

27

Presentation 12
LARVAL FEEDS RESEARCH IN THE UNITED STATESMichael B. Rust
Aquaculture Program, Northwest Fisheries Science Center, 2725 Montlake Blvd E,
Seattle, WA, 98102 USA
E-mail: mike.rust at noaa.gov

This presentation will examine the various types of microparticulate diets currently
being developed in the United States, several new tools to help improvemicroparticulate diets and some international effort to improve collaboration in thisfield. I will also spend the last few minutes on my thoughts on feeds for larval eel.
Following on the pioneering work of Kanazawa and co-workers in Japan, there are threelabs in the US currently working on a new generation of larval feeds. These labs havefocused attention on a variety of microparticle types and combinations of these particles
which are termed complex microparticles. In addition our lab has focused attention on
developing tools that can be used to better test microparticulate diets. So far three tools
have been developed. The first one allows marking diets and live feeds so that
consumption and diet preference can be directly measured. The second one adapts theapparent digestibility methods used by fish nutritionists worldwide on larger fish tosmall larvae. The third method measures mass leaching in real time from microparticle
diets. At the end of this presentation I will defend the idea that eel larvae are parasites
and give my thought about how to test this hypothesis and potential improve larval
nutrition for this important and delicious species.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Larval feeds research in the united states. Michael B. Rust, Aquaculture Program,Northwest Fisheries Science Center, 2725 Montlake Blvd E, Seattle, WA, 98102 USA
Barrows, F. T and W. A. Lellis, 2006. Effect of Diet Processing Method and Ingredient
Substitution on Feed Characteristics and Survival of Larval Walleye, Sander vitreus.
Journal of the World Aquaculture Society, Vol 37:2, 155-160.
Details of producing two types of agglomerated microparticulate diets and theresulting performance of these diets when fed to larval walleye are presented. Dr.
Barrow’s has numerous other papers dealing with larval fish diets and nutrition.
2) Cook, M.A., Johnson, R.B., Nicklason, P.N., Rust, M.B. 2008. Marking live feeds
with inert metal oxides for fish larvae feeding and nutrition studies, Aquaculture
Research.
The authors developed a simple and accurate method to mark live andmicroparticulate feeds in order to quantify consumption. This method can be used to
optimize a variety of conditions in order to maximize feeing success. The method also isthe first step toward determining apparent digestibility in fish larvae.
283) Johnson, R.B., Nicklason, P.N., Cook, M.A., Rust, M.B. in press, Determination of
apparent protein digestibility of live Artemia and a microparticulate diet in 8-week-old
Atlantic cod Gadus morhua larvae. Aquaculture.
The study of larval nutrition has been hampered by a lack of tools to measure
diet performance. In larger fish, one of the key tools is the determination of apparent
digestibility. This paper for the first time, adapts and tests a procedure for determinationof apparent digestibility to larval fish. In this case, the cod are pre-metamorphic, but theauthors are in the process of extending the method back as far as possible to the first
feeding stage.
4) Langdon, C., Nordgreen, A., Hawkyard, M., Hamre, K. in press, Evaluation of wax
spray beads for delivery of low-molecular weight, water-soluble nutrients andantibiotics to Artemia. Aquaculture.
The latest work from Dr. Langdon’s lab describes production and use of lipid
walled microcapsules. Numerous authors have demonstrated that low-molecular weight,
highly soluble nutrients leach within seconds of contact with water for the majority of
microparticle types. Lipid walled beads are one of the few technologies that is capableof retaining highly soluble nutrients. This paper uses the lipid walled microcapsules todeliver highly soluble nutrients to enrich Artemia.
5) Nicklason, P.N., Johnson, R.B. in press, Real-time measurement of protein leachingfrom micro-particulate larval fish feeds, Aquaculture Research.
This paper describes a relatively inexpensive, quick and simple method tomeasure the leaching qualities of microparticulate diets produced by different processesor treated differently. This method could help to better define the physicalcharacteristics of diets used across labs, experiments and species.
6) Finn, R. N. and B. G. Kapoor II (editors). 2008. Fish Larval Physiology. Science
Publishers, Enfield, NH, 724 pages.
Treatise on various aspects on the physiology of larval fishes necessary forunderstanding the underlying biology as it may relate to larval nutrition. Written by 38international experts in 20 chapters and seven sections. Sections address ontogeny,
respiration and homeostasis, nutrition and energy, sensory physiology, movement,
control and defense and functional changes in form.

29

Presentation 13
AQUACULTURE FEED AND SEAFOOD QUALITY
Cheng-Sheng Lee
Center for Tropical and Subtropical Aquaculture, c/o The Oceanic Institute, 41-202
Kalanianaole Hwy. Waimanalo, HI 96795
E-mail: cslee at oceanicinstitute.org

Seafood has been an important additional animal protein source in many less developed
countries. Contribution of seafood from aquaculture has been increasing annually overthe past decades as the harvest from capture fisheries decreased. According to FAO
projection, aquaculture has to continually increase to meet the increasing future demandof seafood. The challenges for aquaculture in the 21st century will include not only
quantity to meet the increasing demand but also quality for health consideration. The
consumers want to have safe and nutritious seafood.
Seafood quality has broad definition from texture, taste, nutritional value tosafety etc. Unlike the seafood harvested from the sea, the quality of aquaculture
products can be controlled by many factors from the production phase to the dinnertable. This report will only discuss factors such as pond management and feeding
contribute to the change of seafood quality.
Since farmed fish feed on the food provided by farmers, the final
nutrients/chemical compositions can be manipulated in farmed fish through feeding.
External appearance such as coloration of fillet can be determined by Astaxanthin
contents in diet. Replacement of fish meal and oil by vegetable protein and oils orchange of oil content in fish feed will change odor and fatty acid composition such as
Polyunsaturated fatty acid (PUFA) levels and consequently, taste in farmed fish. Muscle
lipid content affects the texture of fish as well. The decontamination procedures andenvironmental monitoring during the farming practice may also be a possible means of
reducing some contaminant levels. This presentation is to overview the presentation at
the workshop “Seafood Quality and Aquaculture” held in October, 2007 in Hawaii.
Exchange the means of manipulating the final nutrient composition or boosting a
selected chemical component will increase the product value and benefit the consumers
and producers.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Andersen, U.B., M.S. Thomassen & A.M.B. Rora. (1997) Texture properties of
farmed rainbow trout (Oncorhynchus mykiss): Effects of diet, muscle fat content andtime of storage on ice. Journal of the Science of Food and Agriculture 3, 347-353.
2) Bell, J.G., McEvoy, J., Tocher,D.R., McGhee F., Campbell, P.J. and Sargent, J.R.
2001. Replacement of fish oil with rapeseed oil in diets of Atlantic salmon (Salmosalar) affects tissue lipid compositions and heptocyte fatty acid metabolism. J. Nutr.
30

131: 1535-1543.
3) Boggio, S. M., Hardy, R.W., Babbitt, J.K. and Brannon, E.L. 1985. The influence of
dietary lipid source and alpha-tocopheryl acetate level on product quality of rainbow
trout (Salmo gairdneri). Aquaculture 51, 13-24.
4) Brodtkorb, B.T., G. Rosenlund & O. Lie. (1997) Effects of dietary levels of 20:5n-3
and 22:6n-3 on tissue lipid composition in juvenile Atlantic salmon, Salmo salar, withemphasis on brain and eye. Aquaculture Nutrition 3, 175-187.
5) Cowey, C.B., Adron, J.W., Hardy, R., Smith, J.G.M. and Walton, M.J. 1979.
Utilization by rainbow trout of diets containing partially rendered hide fleshings.
Aquaculture 16: 199-209.
6) De Francesco, M., Giuliana, P., Medale, F., Lupi, P., Kaushik, S. and Poli, B.M. 2004.
Effect of long-term feeding with a plant protein mixture based diet on growth andbody/fillet quality traits of large rainbow trout (Oncorhynchus mykiss). Aquaculture
236: 413-429.
7) Dosanjh, B.S., D.A. Higgs, D.J. McKenzie, D.J. Randall, J.G. Eales, N. Rowshandeli,M. Rowshandeli & G. Deacon. (1998) Influence of dietary blends of menhaden oil andcanola oil on growth, muscle lipid composition, and thyroidal status of Atlantic salmon(Salmo salar) in sea water. Fish Physiology and Biochemistry 2, 123-134.
8) Dupree, H.K., Gauglitz, E.J. and Houle, C.R. 1979. Effects of dietary lipids on thegrowth and acceptability (flavour) of channel catfish (Ictalurus punctatus). Pages
87-103 in J.E. Halver and K. Tiews, editors. Finfish Nutrition and Fishfeeds Technology,
Heenemann, Berlin.
9) Grisdale-Helland, B. and Helland, S.J. 1997. Replacement of protein by fat andcarbohydrate diets for Atlantic salmon (Salmo salar) at the end of the freshwater stage.
Aquaculture 152: 167-180.
10) Halver, J.E. 2002. The Vitamins. Pp. 613-141 in J.E. Halver and R.W.Hardy, editors,
Fish Nutrition, 3rd Edition. Academic Press Inc., New York, NY.
11) Hamre, K. & O. Lie. (1997) Retained levels of dietary α-, γ- and δ-tocopherols intissues and body fluids of Atlantic salmon (Salmo salar L.). Aquaculture Nutrition 3,
99-107.
12) Hardy, R.W., Scott, T.M. and Harrell, L.W. 1987. Replacement of herring oil withmenhaden oil, soybean oil, or tallow in the diets of Atlantic salmon raised in marine
net-pens. Aquaculture 62: 267-277.
13) Hatae, K., A. Tobimatsu, M. Takeyama & J.J. Matsumoto. (1986) Contribution of
the connective tissues on the texture differences of various fish species. Bulletin of31

Japanese Society of Scientific Fisheries 52, 2001-2007.
14) Hites, R.A., Foran, J.A., Carpenter, D.O., Hamilton, M.C., Knuth, B.A., andSchwager, S.J. 2004. Global assessment of organic contaminants in farmed salmon.
Science 303: 226-229. Available at
http://www.sciencemag.org/cgi/content/full/303/5655/226.
15) Horvli, O., L. Aksnes & O. Lie. (1998) Tissue distribution of vitamin D3 in Atlantic
salmon (Salmo salar): Effect of dietary level. Aquaculture Nutrition 4, 89-97.
16) Ikonomou, M.G., Higgs, D.A., Gibbs, M., Oakes, J., Skura, B., McKinley, S., Balfry,
S.K., Jones, S., Withler, R. and Dubetz, C. 2007. Flesh quality of market-size farmed
and wild British Columbia salmon. Environ. Sci. & Techn. 41: 437-443. Abstract
available at
http://pubs.acs.org/cgi-bin/abstract.cgi/esthag/2007/41/i02/abs/es060409+.html.
17) Jobling, M. 2003. Do changes in Atlantic salmon, Salmo salar L., fillet fatty acids
following a dietary switch represent wash-out or dilution? Test of a dilution model andits application. Aqua. Res. 34(13): 1215-1221.
18) Jobling, M. and Johansen, S.J.S. 2003. Fat distribution in Atlantic salmon Salmosalar L. in relation to body size and feeding regime. Aqua. Res. 34(4): 311-316.
19) Lie, O. 2001. Flesh quality- the role of nutrition. Aqua. Res. 32(Suppl. 1): 341-348.
20) Mozaffarian, D. and Rimm, E.B. 2006. Fish intake, contaminants, and humanhealth: Evaluating the risks and the benefits. J. Amer. Med. Assn. 296: 1885-1899.
Abstract available at http://jama.ama-assn.org/cgi/content/short/296/15/1885.
21) Nakagawa, H. and Montgomery, W.L. 2007. Algae. Pp. 133-167 in H. Nakagawa, M.
Sato, and D.M Gatlin III, editors. Dietary supplements for the health and quality ofcultured fish. CAB International, Oxon, UK22) Nakagawa, H., Sato, M. and Gatlin III, D.M., editors. 2007. Dietary supplements forthe health and quality of cultured fish. CAB International, Oxon, UK. 244 pp.
23) National Research Council (NRC) (1993), Nutrient Requirements of Fish. National
Academy Press, Washington, D.C. 114 pp.
24) Nettleton, J.A. and Exler, J. 1992. Nutrients in wild and farmed fish and shellfish. J.
Food Sci. 57(2): 257-260.
25) O’Keefe, T.M. and Noble, R. 1978. Storage stability of channel catfish (Ictaluruspunctatis) in relation to dietary level of α–tocopherol. J. Fish. Res. Bd. Canada 35:
457-460.
3226) O’Neill, S.M., West, J.E. and Hoeman, J.C. 1998. Spatial trends in the concentrationof polychlorinated biphenyls (PCBs) in chinook (Oncorhynchus tshawytscha) and coho
salmon (O. kisutch) in Puget Sound and factors affecting PCB accumulation: resultsfrom the Puget Sound monitoring program.
http://www.psat.wa.gov/Publications/98_proceedings/pdfs/2b_oneill.pdf.
27) Sato, K., R. Yoshinaka, M. Sato & Y. Shimizu. (1986) Callagen content in themuscle of fishes in association with their swimming movement and meat texture.
Bulletin of Japanese Society of Scientific Fisheries 52, 1595-1600.
28) Sigurgisladottir, S., C.C. Parrish, R.G. Ackman & S.P. Lall. (1994b) Tocopherol
deposition in the muscle of Atlantic salmon (Salmo salar). Journal of Food Science 59,
256-259.
29) Shearer, K.D. 1994. Factors affecting the proximate composition of cultured fisheswith emphasis on salmonids. Aquaculture 199(11): 63-88.

33

Presentation 14
QUALITY CONTROL OF CULTURED FISH BY FEED SUPPLEMENTS
Heisuke Nakagawa
Emeritus Professor, Hiroshima University, Hiroshima 739-8528, Japan
E-mail: naka1524 at enjoy.ne.jp

Advancements in fish nutrition have progressed considerably with regard to growth
performance. However, much concern has arisen over the quality of fish flesh andhealth in aquaculture. In response, a variety of feed supplements have been investigatedwith respect to their effects on sensory evaluation, protein metabolism, lipid metabolism,
vitality and disease resistance of cultured fish.
Regular ingestion of substances by wild fish suggests positive function as feed
supplements. The importance of feed supplements was determined in substances that arefound in digestive tracts of wild fish. For example, a variety of wild fishes ingest algae,
including some primarily carnivorous fishes. Crustaceans are important and often the
major food organisms of larval fish and contain considerable amount of chitin. A small
amount of medium-chain fatty acids was found in the stomach content of wild fish.
Therefore, the substance was supplemented to the composed diet of ayu (Plecoglossus
altivelis), red seabream (Pagrus major) and black seabream (Acanthopagrus shlegelii).
The dietary supplements commonly improved physiological condition and normalized
metabolism without depression of growth performance.
Whereas incorporated level of essential nutrients in cultured fish was found to be
fairly low in comparing with those of wild fish, effects of fortification of essential
nutrients such as EPA, DHA and vitamin C were discussed in terms of the above pointof view.
Recently application of new technologies such as molecular biology has
approached to evaluate fish quality and to confirm the effect of feed supplements.
Therefore, some examples of effect of feed supplements determined by gene expression
technique are introduced.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Ji H., A.D. Om, T. Umino, T. Yoshimatsu, M. Hayashi, H. Nakagawa, M. Asano, and
A. Nakagawa 2005. Effect of dietary laurate on lipid accumulation and vitality injuvenile Pagrus major. J. Fish. China, Vol. 29: Pp 804-810.
Based on the fact that medium-chain triglycerides suppress accumulation ofreserved lipids in mammals and some fish species, the effect of dietary laurate on lipid
accumulation and vitality was further examined in red seabream. Ethyl laurate (0.5%)
supplemented to the diet was fed to zero-year fish (0.3 g) for 47 days. While dietarylaurate did not influence growth performance, increasing of muscle ratio and decreasingof liver weight were observed. The dietary laurate led to reducing liver triglycerides,
intraperitoneal fat body ratio, adipocyte diameter, and liver lipid. In addition, DHA,
34

functioning as a physiologically active nutrient, tended to increase in the muscle, liver,heart, brain, and eyes. Dietary laurate activated resistance against air-dipping and liverfunction, as seen the case of dietary DHA fortification. It is concluded that dietarylaurate could depress lipid accumulation and improve vitality, while the influence of
laurate on the tissue DHA deposition might relate to its efficacy.
2) Nakagawa H., M Sato, and D.M. Gatlin III (eds.) 2007. Dietary Supplements for the
Health and Quality of Cultured Fish, 1st ed. CABI Publisher, UK, p. 133-167 and
168-177.
The book addressed current information on the effects of non-macronutrients suchas micronutrients and the efficacious substances from plants, animals, and bacteria, withregard to quality and health of cultured fish. Reports of the nutritional merits of various
substances are based on evidence available in published scientific literature of from yet
unpublished data presented at scientific meetings or acquired as personalcommunications from researchers. Although some feed supplements have been used infish culture without scientific evidence to support their efficacy, the book focuses onsubstances that have been experientially assessed by fish culturists of proven to havebeneficial effects in mammals.
The chapters of the book deal with four parts, as follows;
I. How to evaluate and assess the quality
II. Essential nutrients (amino acids, fatty acids, vitamins, minerals)
III. Feed supplements originating from organisms(microorganisms, terrestrial plants, algae, chitin, plant saponins, nucleotides)
IV. Nutrigenomics, Economics, and food safety

35

Presentation 15
PROTEIN PRODUCTION ADVANTAGES IN THE FACE OF INCREASING
FEED COSTS: IDENTIFYING OPPORTUNITIES WITHIN THE
AQUACULTURE INDUSTRY
Gina Louise Shamshak and James L. Anderson
University of Rhode Island, 1 Greenhouse Road, Kingston, Rhode Island 02882 USA
E-mail: shamshakg at yahoo.com

This presentation will provide an overview of the trends in the global production of raw
feed materials. Recent trends impacting the production and prices of key raw feed
ingredients will be highlighted and discussed. This will include a discussion of thecurrent and future trends in production and prices for both agricultural sources of key
feed ingredients (soybeans, corn) as well as marine based (fishmeal, fish oil) sources.
The focus will then turn to a discussion of feed ingredients and feed costs as they relate
to the production of key proteins, including both agriculture and aquaculture sources of
production (e.g. beef, chicken, pigs, salmon, and catfish). It will then be argued that theaquaculture sector is in a strong position to provide quality protein at competitive prices
despite increases in raw feed prices. A critical factor will be the degree to which theaquaculture sector, and further, certain species within the aquaculture sector, cansubstitute across key feed ingredients. Substitution gives farmers the opportunity to
reduce feed costs in the face of rising feed prices by allowing them to find feedcombinations that are cost effective and also nutritionally sufficient. Thus, given thepotential for substitution within the aquaculture sector, producers have the opportunity
to manage feed costs in a manner that maintains production of quality protein atcompetitive prices.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Offshore Aquaculture in the United States: Economic Considerations, Implications
and Opportunities. National Oceanic and Atmospheric Administration. National
Aquaculture Program. Silver Spring, MD. July 2008 (Pre-Publication Copy)
http://aquaculture.noaa.gov/news/econ.html
The report by leading fisheries and resource economists and business experts,
Offshore Aquaculture in the United States: Economic Considerations, Implications &Opportunities, examined a range of topics on the industry’s potential and found that asignificant domestic offshore aquaculture industry could develop and be successful overthe next 20 years with a clear regulatory framework. One chapter in particular (Chapter4) examines the future of aquaculture feeds and feed costs by examining the role of fishmeal and fish oil. The authors suggest that the limited supply of fish meal and fish oil is
not likely to be a constraint on the expansion of U.S. aquaculture as alternative feed
ingredients from soybeans and other plants, algae, yeasts, fish processing waste, andother products are being developed.
362) Jackson, Andrew. 2007. Challenges and Opportunities for the Fishmeal and Fish oil
Industry. Feed Technology Update. Volume 2, Issue 1. January 2007.
http://www.aquafeed.com/newsletter_pdfs/nl_000245.pdf
This paper first examines the current state of the global fish meal and fish oil
industry and then examines the claim that fish meal and fish oil might limit the future
growth of the aquaculture industry. The author asserts that fishmeal and fish oil will
become more strategic dietary ingredients, rather than limiting the growth of theaquaculture industry.
3) Kristofersson, Dadi. and James L. Anderson. 2006. “Is There a Relationship between
Fisheries and Farming? Interdependence of Fisheries, Animal Production andAquaculture” Marine Policy, Vol. 30, No. 6: 721-72
These authors examine the role of fishmeal as a critically important feed
ingredient for use in animal (e.g., poultry and pork) and aquaculture production (e.g.,salmon, trout, and shrimp). Their research suggests that since 1998 market for fishmeal
has changed. An important explanation is likely to be the increasing use of specialized
feed formulations in the pork, poultry and aquaculture sectors. It is clear that growth inaquaculture production alone is insufficient to explain the structural change since
growth in the use of fishmeal in aquaculture has slowed considerably despite thecontinued rapid growth in global aquaculture sector. The increasing relative price offishmeal is likely to increase costs for animal producers, act as a stimulus for innovationand have considerable implications for the management of pelagic fisheries.

37

Presentation 16
AVAILABILITY OF GENE-MODIFIED FEED INGREDIENTS TO FISH
Shuichi Satoh* and Pitchaya Chainark
Laboratory of Fish Nutrition, Tokyo University of Marine Science & Technology, Tokyo
108-8477, Japan
E-mail: ssatoh at kaiyodai.ac.jp

The feeding studies were conducted to assemble the information on the availability ofgene-modified (GM) feed ingredient for rainbow trout, using diet containing GM
defatted soybean meal (SBM) as an alternative protein source to provide and ensure
good protein accessibility and product safety. The utilization of genetically modified
defatted soybean meal (GM SBM) as feed by rainbow trout was investigated, incomparison with non genetically modified defatted soybean meal (non-GM SBM). The
nutrient utilization showed that there was no significant difference in growth and feed
performance between GM and non-GM SBM groups in 12 week feeding experiment.
However, the cauliflower mosaic virus 35S promoter fragment of the GM SBM wasdetected in the muscle of fish receiving GM SBM diet by Nested-PCR. Additionally, thepromoter fragment vanished by the 5th day after changing the diet to non-GM diet.
Subsequently, the study was carried out to examine the degradation and the possible
carryover of foreign DNA fragment by means of measuring it from transgenic plant andhost plant contained in GM or non-GM SBM and evaluate the safety for fish. These
foreign DNA fragments were not completely degraded in stomach and intestine andmight be taken up into organ via the garsrointestinal (GI) tract. However, foreign DNA
was not detected after the withdrawal period. Judging from these findings, the novel
feed ingredients derived from GM SBM could be considered as having equivalentnutritional quality and verifying the safety as feed ingredient.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) P. Chainark, S. Satoh, T. Hino, V. Kiron, I. Hirono, and T. Aoki, 2006. Availability ofgenetically modified soybean meal in rainbow trout (Oncorhynchus mykiss) diets.
Fisheries Science, 72 (5), 1072-1078.
The use of genetically modified(GM) defatted soybean meal as rainbow trout
feed was investigated, in comparison with non-GM SBM. Both SBMs were included at
levels of approximately 15 and 30% in four diets. The diets were fed to juvenile fish for
12 weeks. The nutrient use showed that there were no significant differences in growthand feed performance between GM and non-GM SBM groups at both inclusion levels at
the end of 12th week. The cauliflower mosaic virus 35S promoter fragment of the GM
SBM was detected in the muscle of fish receiving GM SBM diet by nested PCR.
Additionally, the promoter fragment was not detected by the fifth day after changing thediet to non-GM. Conversely, the promoter fragment was not detected from fish fed thenon-GM SBM diet. The results demonstrated that the availability of protein in GM
38

SBM was similar to that of non-GM SBM, and the promoter fragments found in themuscle of fish were not detectable after changing the diet to non-GM, verifying theavailability of the GM SBM in rainbow trout feed.
2) P. Chainark, S. Satoh, I. Hirono, T. Aoki, and M. Endo, 2008. Availability ofgenetically modified feed ingredient II – Investigations of ingested foreign DNA in
rainbow trout, Oncorhynchus mykiss. Fisheries Science, 74 (2), 380-390.
The foreign DNA fragment from genetically modified defatted soybean meal
(GM SBM) in rainbow trout was traced by nested-PCR and located by in situ
hybridization. GM or non-GM SBM formulated diet (42% protein) was fed to fish (Av.
Wt. 50.5 g) for 2 weeks. The degradation results showed that the cauliflower mosaic
virus 35S promoter (220 bp) fragment was detected in the contents of digestive system
only in fish fed GM SBM diet, and there was not detected on the 3 rd day after changing
the diet to non-GM SBM diet. For the possible transferal results, the promoter fragment
was detected in the leukocyte, head kidney and muscle of only fish fed GM SBM diet,afterward it was not detected on the 5th day after changing the diet to non-GM SBM diet.
These results suggested that foreign DNA fragment was not completely degraded andmight be taken up into organ via the gastrointestinal tract. However, foreign DNA was
not detected after the withdrawal period. Thus, the data might be considered that uptakeof DNA from GM SBM might not remain in the tissues of the fish fed GM SBM diet.

39

Presentation 17
QUANTIFICATION OF WASTE FEED AND FISH FECES USING STABLE
CARBON AND NITROGEN ISOTOPES
Hisashi Yokoyama*, Yuka Ishihi, Katsuyuki Abo, and Toshinori Takashi
Aquaculture Systems Division, National Research Institute of Aquaculture, FRA, Mie
516-0193, Japan
E-mail address: hyoko at fra.affrc.go.jp

For improving the economic and environmental sustainability of fish farming, it isnecessary to optimize feeding regimes and to decrease waste feed. We developed amethod to quantify waste feed and fecal matter in sediment trap materials and sediments
beside and beneath fish cages using stable carbon and nitrogen isotope ratios. The
contribution ratio of three sources (waste feed, fecal matter and natural organic matter)in sedimentary organic matter in and around a red sea bream (Pagrus major) farm inGokasho Bay, central Japan, was estimated using an isotopic mixing model. The result
showed that waste feed (29% in the bulk organic matter) exceeded fecal matter (12%) in
the fish farm area, suggesting the overfeeding in this farm. Then we monitored thegrowth and mortality of red sea bream, C and N fluxes and sediment chemistry at twocommercial fish cages; cage 1 (conventional satiation feeding) and cage 2 (restricted
feeding: same feeding frequency but 18% reduction in the feed amount) for 276 days.
The restricted feeding achieved normal growth, increased feed conversion efficiency
and reduced mortality of red sea bream and reduced contents of waste feed-derived
organic matter in the sediment. Determining the relative amount of waste feed and fecal
matter in settling and sedimentary organic matter is effective for the evaluation of the
optimum ration level from the viewpoint of the minimization of waste feed.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Yokoyama, H., K. Abo, and Y. Ishihi. 2006. Quantifying aquaculture-derived organic
matter in the sediment in and around a coastal fish farm using stable carbon andnitrogen isotope ratios. Aquaculture, Vol: 254. Pp 411-425.
The stable carbon and nitrogen isotope ratios of the sedimentary organic matter(SOM) collected from 41 stations in and around a coastal fish farm in Japan were
measured to quantify aquaculture-derived organic matter in the sediment. SOM in thefish-farm area (within 30 m from the edge of cages) is characterized by its reduced δ13C
(mean ⊿δ13C = –0.4‰) and enriched δ15N (mean ⊿δ15N = +0.9‰) values, whichreflect the deposition of C3-plant-derived and fish-derived elements, respectively.
Compositions of waste feed (WF) and fecal matter (FM) in SOM at each station wasdetermined based on the isotopic compositions of feed (δ13C = –20.2‰, δ15N = 9.7‰),
fish feces (δ13C = –24.3‰, δ15N = 6.3‰) and marine organic matter in the sediment(δ13C = –19.9‰, δ15N = 5.5‰). The sediment in the fish-farm area was characterized byhigh WF and FM ratios in SOM (28.8 and 11.9%). As the distance from the fish cages
40

increased, aquaculture-derived organic matter decreased exponentially. The spatial
extent of waste dispersal extended to an area up to 300 m, whereas dissolved oxygen of
the bottom water and acid volatile sulfides in the sediment were affected even at stations
600 m away from the fish farm. There was a significant negative relationship between
the aquaculture-derived nitrogen content in the sediment and the mean current velocity,suggesting that areas (water depth = ca. 18 m) where the near-bed current velocity is >8cm/s will not receive excessive accumulation of organic wastes.
2) Yokoyama, H., and Y. Ishihi. 2007. Variation in food sources of the macrobenthos
along a land-sea transect : a stable isotope study. Marine Ecology Progress Series,
Vol:346. Pp127-141.
To evaluate the relative importance for the macrobenthos of possible foodsources, including riverine particulate organic matter, reeds, benthic microalgae,
seaweeds, coastal phytoplankton and aquaculture-derived matter, over 194 macrofaunal
species were collected from 9 intertidal and subtidal stations along the axis of Gokasho
Bay, Japan, and their isotopic compositions analyzed. The δ13C values for terrestrialorganic matter were very different from those of the consumers (>5‰), suggesting anegligible trophic role for this source. Differences in δ15 N between seaweeds andprimary consumers were small, suggesting a minor contribution of seaweeds. The δ13C
values of the majority of consumers fell between coastal phytoplankton and benthic
microalgae, allowing the calculation of the relative contribution of these microalgae tothe diets of consumers. The estimated contribution ratios and the between-sitecomparison of the δ13C value for the same species along the bay axis suggest a shift ofthe main food source from benthic microalgae on the mudflat to coastal phytoplankton
at the subtidal seaward stations. Several consumers collected from the fish-farm area
had enriched δ15N and depleted δ13C values relative to samples of the same species that
occurred outside of the farm area. The δ13C values of the consumers at the fish farm
were significantly more depleted than those at the other stations. These findings suggestthat the consumers inhabiting the farming area incorporate 15 N-enriched fish meal and13
C-reduced cereals that are major constituents of fish feed.
3) Uede T., and T. Takeuchi. 2007. Postprandial changes in digesta, leftovers and feces,and their carbon and nitrogen contents in cultured red sea breams, Pagrus major.
Aquaculture Science, Vol:55:3. Pp. 409-415.
For establishing a sustainable aquaculture for red sea bream, we examined
postprandial changes in digesta and suspending matter discharged as leftovers and feces.
Gastral digestion for extruded pellet (EP) needed 24 h in the fish weighing 158 g atwater temperature range of 20.3-26.6°C. Enteral digestion for EP almost ceased at about
72 h after feeding. The C/N ratio of suspending matter was similar to EP until 3 h after
feeding, and rapidly rose after then. This suggested that suspending matter until 3 h after
feeding and after then was originated from leftovers and feces, respectively. EP given
was discharged as leftovers and feces ranges of 3.6-15.4 and 4.1-14.7%, respectively.
Moist pellet (MP) given was also discharged as leftovers and feces ranges of 10.1-19.2
and 2.6-10.8%, respectively. Feed intake, leftovers and feces of MP were1.9, 3.5 and 1.5
times greater than those of EP, respectively. Until 3 h after feeding EP, N in leftovers
was at the range of 48.3-57.4% of total N discharged. Thereafter, it is intensely
41

recommended that preferable feeding restriction with EP is effective for conducting
nitrogen load reduction and sustainable aquaculture for red sea bream. [In Japanese withEnglish summary]42

Presentation 18
MEASUREMENT OF SWIMMING BEHAVIOR AND DIGESTIVE PROCESSOF CULTURED FISH
Toshinori Takashi1*, Wataru Sakamoto2, Kubo Toshihiko2, Yasushi Mitsunaga3, Osamu
Murata21

Aquaculture Systems Division, National Research Institute of Aquaculture, FRA, Mie
516-0193, Japan
2
Fisheries Laboratory, Kinki University, Wakayama 649-2211, Japan
3
Faculty of Agriculture, Kinki University, Nara 631-8505, Japan
E-mail: ttakasi at fra.affrc.go.jp

The biotelemetry method, which can measure fish behavior and physiologicalconditions such as body temperature and cardiac beat, is a useful tool for aquaculture to
manage health and feeding cultured fish. In this study, we introduce two topics which
are about monitoring of cultured Pacific bluefin tuna (PBT) using micro data-logger. (I)Using the acceleration data logger, swimming depth, tail beat and body angle of PBT
were monitored in two different sizes of net cage. The monitored data indicatedinfluence of net case size on cultured PBT behavior. The swimming behaviors of PBTs
were apparently different between in the large and small net cages. In the small net cage,
the PBT kept their swimming speed slowly, and frequently change the body angle innighttime. In contrast, the PBT in the large net cage swim faster and body angle wasstable. (II) We also monitored abdominal cavity temperature of PBT to estimate basal
physiological condition in a net cage. The PBT maintains higher body temperature inseveral degrees than ambient water temperature. However, the body temperature waschanged by the feeding and temperature fluctuation. The results suggested that theabdominal cavity temperature fluctuation was influenced by the ambient temperature,swimming activity, and food digestion and assimilation.
ANNOTATED BIBLIOGRAPHY OF KEY WORKS
1) Kubo T., W. Sakamoto, O. Murata AND H. Kumai. 2008. Whole-body heat transfer
coefficient and body temperature change of juvenile Pacific bluefin tuna Thunnus
orientalis according to growth. Fisheries Science, Vol.74, pp.995-1004.
Bluefin tuna maintain a higher body temperature than ambient sea water. Body
heat is derived mainly from metabolic heat to elevate and maintain regional bodytemperature that is higher than the ambient, while heat loss is caused by heat transfer
throughout the whole body surface and gills. Retention of high body temperature isthought to differ at each growth stage, so that a larger body mass maintains a higher
body temperature. We evaluated the whole-body heat transfer coefficient, thermal
difference between each tissue and water temperature, and metabolic heat in tissues
during swimming of juvenile bluefin tuna as a function of fork length (FL) using a small
thermometer and a treadmill-type flow tank. A system for maintaining high body43

temperature was well developed in fish with FL greater than 20.0 cm. Whole-body heattransfer coefficient was fitted to a – 0.695 power of mass. Juvenile bluefin tuna showeda transition speed of 3.0 FL/s at which they switched from aerobic to anaerobic motion.

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