United States-Japan Natural Resources (UJNR)
Panel on Aquaculture
41st Scientific Symposium“Advanced Aquaculture Technologies”
October 9-10, 2013
Sapporo, Hokkaido

Annotated Bibliography

Photo: Spawning sea cucumbers (Apostichopus japonicus) induced with “cubifrin” pictured by Dr. K.
Yamano (FRA)UJNR Annotated Bibliography

DEVELOPMENT OF A NEW TYPE OF FISH DIET, NON-FISH MEAL
EXTRUDED-PELLET
Ishida, N
N. Praxhom, Y. Haga, and S. Satoh (2013). Impact of dietary high protein distillers dried grains on aminoacid utilization, growth response, nutritional health status, and waste output in juvenile rainbow trout(Oncorhynchus mykiss)), Aquaculture Nutrition, 19, Issue Supplement s1, 62-71
High protein distillers dried grains (HPDDG) is a co-product of ethanol production that uses
prefractionation technology. A 14-week nutrition trial of HPDDG with rainbow trout (Oncorhynchus
mykiss: initial weight = 7.5–7.7 g fish−1) was conducted to evaluate the effect of replacing corn gluten
meal (CGM) with four levels of HPDDG (0, 50, 100 and 150 g kg−1). HPDDG is higher in crude protein(481 g kg−1, CP) than typical distillers dried grains with soluble (DDGS) (270 g kg −1). We found that 0–150 g kg−1 HPDDG can be used in diets for rainbow trout. Replacement of CGM by HPDDG did notaffect feed utilization efficiency or the health status of fish. The final body weight and phosphorus (P)content of fish increased with increasing HPDDG content in the feed. Although increasing dietary
HPDDG levels resulted in a reduction in lysine (Lys) and methionine (Met) digestibility, the digestible CP,
including Lys and Met, in the experimental diets remained high (above 90%). Better utilization of CP and
P by fish fed dietary HPDDG resulted in less waste excretion by these fish than by those fed the control
diet. HPDDG may be considered as an alternative protein source for aquaculture feeds.

S. Boonyoung, Y. Haga, S. Satoh (2013). Preliminary study on effects on methionine hydroxyl analog andtaurine supplementation in a soy protein concentrate-based diet on the biological performance and aminoacid composition of rainbow trout [Oncorhynchus mykiss (Walbaum)]. Aquaculture Research, 44, Issue

9, 1339-1347
A feeding trial was conducted on the effects of methionine hydroxy analog (MHA) and taurine
supplementation in diets with high levels of soy protein concentrate (SPC) on the growth performance andamino acid composition of rainbow trout, Oncorhynchus mykiss (Walbaum) comparing with fish mealbased diet. The control diet had 520 g kg−1 fish meal. In the methionine deficient diets (5.1 g kg−1), fishmeal was replaced by 490 g kg−1 of the SPC in the SPC49 diet. The SPC49 diet was supplemented witheither MHA (6 g kg−1) only or a combination of MHA and taurine (2 g kg−1). Fish were fed isoproteic(460 g kg−1) and isolipidic (130 g kg−1) diets for 12 weeks. Growth performance (i.e. weight, feed
conversion ratio, and thermal-unit growth coefficient) was inferior in fish fed the SPC49 diet. MHA
supplementation improved growth performance (P < 0.05). No difference was observed when taurine wasadded to the SPC49 and MHA diet (P > 0.05). Whole-body taurine contents increased with taurine
supplementation, whereas plasma methionine increased with MHA supplementation (P < 0.05). In

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conclusion, the substitution of fish meal with SPC supplemented with MHA did not negatively impact
growth, and the addition of taurine did not improve growth performance in rainbow trout.

Md. S. A. Sarker, S. Satoh, S. Kamata, K., Haga, Y., and Y. Yamamoto (2012). Partial replacement offishmeal with plant protein sources using organic acids to practical diets for juvenile yellowtail, Seriola
quinqueradiata, Aquaculture Nutrition, 18, 81-89
An experiment was conducted to investigate the effects of phosphorus (P), citric acid (CA) andformic acid (FA) supplementation on growth and loading of nitrogen (N) and phosphorus (P) in juvenile
yellowtail fed fish meal (FM) and alternate plant protein (APP) diets. Six diets designated as F49 (490 g
kg−1 FM-based), F49 + P (490 g kg−1 FM with 5 g kg−1 inorganic P), F31 + CA (310 g kg−1 FM containing
200 g kg−1 APP with 5 g kg−1 CA), F31 + FA (310 g kg−1 FM having 200 g kg−1 APP with 4 g kg−1 FA),
F23 + CA (230 g kg−1 FM containing 300 g kg−1 APP with 5 g kg−1 CA) and F23 + FA (230 g kg−1 FMhaving 300 g kg−1 APP with 4 g kg−1 FA) were formulated. Yellowtails were fed one of the diets for 12
weeks under on-site conditions at water temperature 22.0–27.0 °C. F49 + P gave the best growth, while
F23 + CA the lowest. Specific growth rate and feed conversion ratio of F31 + CA were similar with
control group. Addition of CA to APP diets significantly increased retention of P; hence, its excretion was
lowered. The results of this study suggest that FM could be replaced up to 70% with the addition of 5 g
kg−1 CA to a low P-containing plant protein sources diet without inorganic P supplementation in juvenile
yellowtail diets, which in turn would enable a reduction in environmental pollution from aquafeeds.
Md. S. A. Sarker, S. Satoh, S. Kamata, K., Haga, Y., and Y. Yamamoto (2012). Supplementation effect oforganic acids and/or lipid to plant protein based diets on juvenile yellowtail, Seriola quinqueradiata
Temminck et Schlegel 1845, growth and, nitrogen and phosphorus excretion, Aquaculture Research, 43,

Issue 4, 538-545

A feeding experiment was conducted to investigate the effect of organic acids and/or
lipid supplementation on growth, utilization and environmental loading of nitrogen (N) andphosphorus (P) in juvenile yellowtail fed fishmeal (FM) and plant protein (PP) diets. Six
diets as FM (FM-based), FM+P (FM with inorganic P), FM+L (FM with lipid), PP+CA (PPwith citric acid), PP+L+CA (PP with lipid and citric acid) and PP+L+FA (PP with formicacid) were formulated. Yellowtails were fed each of the diets in duplicate groups; once a day,
6 days a week to near satiation at water temperature 19.0–25.0 °C for 16 weeks. Fishmeal
with inorganic P gave the best growth while PP+L+FA the lowest. However, growthincreased in PP+CA and PP+L+CA. Addition of lipid significantly increased N and P
retention resulting in significant reduction in N and P excretion. Citric acid and FA
supplementation to PP diets also increased retention of P; hence, its excretion was lowered.
Thus, CA, FA and lipid in juvenile yellowtail diets can help to partially replace FM with PP
sources and reduce inorganic P use to minimize environmental loading from aquafeeds.
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AVAILABILITY OF FISHERIES BY-PRODUCT MATERIALS WITH CADMIUMREMOVAL TREATMENT AS A FEED INGREDIENT FOR FINGERLING BLACK
ROCKFISH SEBASTES SCHLEGELI
Satoh, N
Satoh N, Nobuta S, Wakasugi M, Satoh S, Takeuchi T (2013). Availability of squid viscera meal withcadmium removal treatment as a feed ingredient for fingerling black rockfish Sebastes schlegeli. Fisheries
Science 79: 259-267
Two 12-week feeding experiments were conducted to evaluate the nutritional value of squid viscera
meal with cadmium removal treatment (dCSVM), which contained 1.5-2.0 mg/kg cadmium, as analternative protein source to sardine meal in diets for fingerling black rockfish Sebastes schlegeli. Initialmean body weights in experiments 1 and 2 were 23.0 and 6.4 g, and the replacement rates of sardine meal
with dCSVM in the test diets were 20-80 % and 30-60 %, respectively. In experiment 1, inclusion ofdCSVM at 40 % and higher replacement rates of sardine meal retarded the growth of fish due partly toinferior dCSVM protein digestibility. The cadmium (Cd) concentrations in fish muscle of all treatmentgroups were below the detection limit (<0.1 mg/kg dry matter). In experiment 2 using dCSVM containing
Cd at a lower level and having a higher protein digestibility than experiment 1, no significant differenceswere observed in the growth between the control and 60 % replacement groups, although feed efficiency
gradually decreased with the increase of dCSVM inclusion. These results show that dCSVM is safe anduseful as an alternative protein source and could replace up to 60 % of sardine meal in fingerling black
rockfish diets.

Satoh N, Fujioka T, Nobuta S (2006). Availability of commercial squid viscera meal as diets of fingerling
black rockfish. Nippon Suisan Gakkaishi 72:401-407 (in Japanese with English abstract)
Feeding experiments were conducted to evaluate the growth performance, feed efficiency, total foodintake and heavy metal accumulation (Cadmium and Copper) in fingerling black rockfish Sebastes
schlegeli, fed extruded pellets containing commercial diets supplemented with 0% (control) to 30%
commercial squid viscera meal without Cadmium removal treatment (CSVM) for eight weeks.
Growth performance did not differ significantly in any of the experimental groups, but total foodintake increased in response to increased CSVM. Findings also suggested that diets supplemented with
CSVM promoted fish feeding. However, accumulation of Cadmium and Copper in the liver of fish wasmore apparent in experiment groups fed diets containing 20 or 30 % CSVM. These findings indicate that
CSVM should only be applied at levels of up to10 % in the diets of fingerling black rockfish.

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EFFECT OF FEED INGREDIENTS ON DIGESTIVE ENZYMES SECRETION IN
FISH
Murashita, K
Koji Murashita, Haruhisa Fukada, Ivar Rønnestad, Tadahide Kurokawa, Toshiro Masumoto(2008). Nutrient control of release of pancreatic enzymes in yellowtail (Seriola
quinqueradiata): Involvement of CCK and PY in the regulatory loop. Comparative
Biochemistry and Physiology, Part A 150: 438-443
Cholecystokinin (CCK) and neuropeptide Y (NPY)-related peptides are the key
regulators of pancreatic enzyme secretion in vertebrates. CCK stimulates enzyme secretion
whereas peptide Y (PY), a NPY-related peptide, plays an antagonistic role to that of CCK. In
fish, very little is known about the effects of different nutrients on the synthesis of CCK and
PY in the digestive tract, and the mechanism by which CCK and PY actually regulate
digestive enzyme secretion is not well understood. In order to determine stimulating effects
of different nutrients on the synthesis of CCK and PY in yellowtail (Seriola quinqueradiata),
CCK and PY mRNA levels in the digestive tract were measured after oral administration of asingle bolus of either phosphate-buffered saline (PBS: control), starch (carbohydrate), casein(protein), oleic acid (fatty acid) or tri-olein (triglyceride). In addition, in order to confirm thesynthesis and secretion of digestive enzymes, the mRNA levels and enzymatic activities ofthree digestive enzymes (lipase, trypsin and amylase) were also analyzed. Casein, oleic acidand tri-olein increased the synthesis of lipase, trypsin and amylase, while starch and PBS did
not affect the activity of any of these enzymes. CCK mRNA levels rose, while PY mRNAlevels were reduced in fish administered casein, oleic acid and tri-olein. These results
suggest that in yellowtail, CCK and PY maintain antagonistic control of pancreatic enzyme
secretion after intake of protein and/or fat.
Takahiro Furutani, Toshiro Masumoto, Haruhisa Fukada (2012). Response ofcholecystokinin and digestive enzyme mRNA levels to various feed ingredients in yellowtail
Seriola quinqueradiata. Fisheries Science 78: 1075-1082
Cholecystokinin (CCK) is the key regulator hormone that stimulates the secretion ofdigestive pancreatic enzymes in vertebrates. In fish, little is known about the mechanism of
induction of CCK in the digestive tract by different feed ingredients. To investigate theresponse of CCK and digestive enzymes to fish feed ingredients in yellowtail Seriola
quinqueradiata, we performed a series of experiments in which we measured the mRNAlevels of CCK, trypsin, and lipase after oral administration of a single bolus of various
ingredients. We administered fish meal and fish oil in experiment 1; high and low
concentrations of fish meal in experiment 2; and five different dietary protein sources (fish4

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meal, soybean meal, soy protein concentrate, corn gluten meal, and glutamic acidfermentation by-products) in experiment 3. In experiments 1 and 3, only fish meal
significantly increased the mRNA level of CCK and digestive enzyme. In experiment 2, ahigh concentration of fish meal [20 % (w/v)] significantly increased the CCK and trypsin
mRNA levels, but a low concentration of fish meal [1 % (w/v)] did not. These results suggestthat high concentrations of fish meal (the protein source in fish feed) has the most potent
effect on stimulation of CCK synthesis and secretion of digestive enzymes in yellowtail.
Takahiro Furutania, Toshiro Masumoto, Haruhisa Fukada (2013). Molecular cloning andtissue distribution of cholecystokinin-1 receptor (CCK-1R) in yellowtail Seriola
quinqueradiata and its response to feeding and in vitro CCK treatment. General andComparative Endocrinology 186: 1-8
In vertebrates, the peptide cholecystokinin (CCK) is one of the most importantneuroregulatory digestive hormones. CCK acts via CCK receptors that are classified into two
subtypes, CCK-1 receptor (CCK-1R; formally CCK-A) and CCK-2 receptor (formally
CCK-B). In particular, the CCK-1R is involved in digestion and is regulated by CCK.
However, very little information is known about CCK-1R in fish. Therefore, we performed
molecular cloning of CCK-1R cDNA from the digestive tract of yellowtail Seriola
quinqueradiata. Phylogenetic tree analysis showed a high sequence identity between thecloned yellowtail CCK receptor cDNA and CCK-1R, which belongs to the CCK-1R cluster.
Furthermore, the expression of yellowtail CCK receptor mRNA was observed in gallbladder,pyloric caeca, and intestines, similarly to CCK-1R mRNA expression in mammals,suggesting that the cloned cDNA is the yellowtail CCK-1R. In in vivo experiments, theCCK-1R mRNA levels increased in the gallbladder and pyloric caeca after feeding, whereas
in vitro, mRNA levels of CCK-1R and digestive enzymes in cultured pyloric caeca increasedby the addition of CCK. These results suggest that CCK-1R plays an important role indigestion stimulated by CCK in yellowtail.

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AN ANALYSIS OF THE CAUSALITY BETWEEN THE MARKET PRICE OF
IMPORTED FISHMEAL AND MARKET PRICE OF MARINE FARMED FISH
Takahashi, Y

Ariji M, Takahara A, Kurata T and Munekiyo M(2002). A study on resource management and market ofqueen crab in Kyoto prefecture: Cointegration test with vector error correction model, Journal of Regional
Fisheries, 43(1) : pp.93-111
In this study, they clarified some useful facts for fishery policy. Maizuru and Central market is notaffected by impulse of other markets, this result indicates policy makers do not have to care for market
intervention if policy makers intervene in Maizuru and Central market. Also, they revealed that impulse
caused by Maizuru market is very strong, and small scale markets around Maizuru are affected by that
impulse. Therefore, soaring queen crab price in Maizuru market cause positive effects to small scale
markets, if fishermen in Maizuru make progress in technological innovations such as high quality
production or market expansion
Ariji M, Kurata T, Sasaki Y and Munekiyo M (2004). Study about clarifying the market circulation andthe price decision factor of the tile fish market in Kyoto, Journal of Regional Fisheries, 45(1) : pp.85-100
In this study, they clarified market circulation and the price decision factor of the tile fish market in
Kyoto and examined about the validity of the introducing the ECR (Efficient Consumer Response) system.
First, they qualitatively examined characteristics of the tile fish and its market and the structure of the tile
fish circulation. Secondly, They analyzed the market circulation and the price decision factor of the tile
fish in Kyoto using analysis of inverse demand function of ECM (Error Correction Model). As a result of
the analysis, it became clear that introducing ECR from a stage of the Central Wholesale Market that the
introduction of ECR in a stage of a retail market was effective against branding.

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INDUCED SPAWNING IN THE SEA CUCUMBER APOSTICHOPUS JAPONICUS
BY NEUROPEPTIDE, CUBIFRIN
Yamano, K
Kato S, Tsurumaru S, Taga M, Yamane T, Shibata Y, Ohno K, Fujiwara A, Yamano K,
Yoshikuni M (2009) Neuronal peptides induce oocyte maturation and gamete spawning ofsea cucumber, Apostichopus japonicus. Dev Biol 326: 169-176
Extracts prepared from tissues containing buccal ring nerve or longitudinal radial nerveof sea cucumber induce oocyte maturation and ovulation from ovarian tissues. We purified
two small peptides, a pentapeptide and a heptapeptide, from the buccal tissues of Japanesecommon sea cucumber, Apostichopus japonicus. Both peptides induced oocyte maturationand gamete spawning. The pentapeptide was identified as NGIWY-amide. This peptide
induced in vitro germinal vesicle breakdown and ovulation of fully- grown oocytes at lessthan 1 pM and in vivo spawning at 10 nM. A synthetic derivative of the pentapeptide,NGLWY-amide, was 10–100 times more potent compared to the natural NGIWY-amide. The
heptapeptide was less potent, inducing ovulation at 1 μM. NGIWY-amide and
NGLWY-amide induced a characteristic spawning behavior when injected into sexuallymatured individuals. Mature eggs artificially spawned were fertilized, and developed
normally and metamorphosed into young sea cucumbers. The details of the production andthe mechanism of action of NGIWY-amide are still unclear, but the high biopotency of thepeptide will aid understanding of the neuronal and hormonal control of reproduction of sea
cucumber.

Fujiwara A, Yamano K, Ohno K, Yoshikuni M (2010) Spawning induced by cubifrin in theJapanese common sea cucumber Apostichopus japonicus. Fish Sci 76: 795–801
The neuropeptide cubifrin-I and its derivative cubifrin-L have recently been
demonstrated as potent substances that induce oocyte maturation in vitro and spawn in theJapanese common sea cucumber Apostichopus japonicus. Here, the reproductive behavior
provoked by injection of cubifrin-L into the body cavity of A. japonicus was examined witha view to the practical application of the peptide for induction of spawning in the hatchery.
Ovarian fragments with oocytes larger than 155 µm in diameter responded to cubifrin-L in
vitro. The in vitro responsiveness of ovarian fragments was well correlated with thespawning success induced in vivo by a cubifrin-L injection. Mature sea cucumbers injectedwith cubifrin-L displayed sequential reproductive behaviors, which comprised climbing theside wall of the tank toward the water surface, waving of the head, and shedding of gametes.
Gamete shedding started about 60 min and 80 min after the injection in males and females,
respectively, and was completed almost simultaneously in the two sexes about 2 hours after
the administration. Repeated injections of cubifrin-L at intervals of about 10 days
successfully induced multiple spawnings in males and females. This study demonstrated thatcubifrin-L is an effective inducer of spawning in Japanese sea cucumber cultivation.

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MASS PRODUCTION OF ARTIFICIAL SEED OF JAPANESE COMMON SEA
CUCUMBER (APOSTICHOPUS JAPONICUS) IN HOKKAIDO, JAPANSakai, Y
Yanagisawa, T.(1998) Aspect of the biology and culture of the sea cucumber.
Sena S. De Silva, ed. Tropical Mariculture, p.291-308, Academic Press, London,
Seed production and releasing technique of Japanese sea cucumber Apostichopus japonicus had
introduced in this paper include economic aspects.

Sakai, Y. and Konda, Y. (2008) Manamako jinkou syubyou no syokugai boujyo giijyutu-ni tuite (Newmass production technique to prevent the predation of artificial juveniles from copepods) Hokusuishi
Dayori 76: 8-14. (in Japanese)
Major two new methods to prevent the predation of Apostichopus japonicus by copepods (Tigriopus
japonicus) increased in the juvenile rearing tank had reported in this paper.

Sakai, Y. (2012) Syubyou seisan to saiabi gyogyou (Artificial seed production and Aquaculture of
Apostichopus japonicus), Takahashi, A. and Okumura, S. (eds). Namakogaku: Seibutu, Sangyo andBunka (Sea Cucumber: Biology, Industries and Culture). p101-114, Naruyamadou, Tokyo.
(in Japanese)
Recent status of mass production technique of A. japonicus includes intermediate culture and future
aspects had summarized.

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IMPROVING THE FOOD QUALITY OF SEA URCHIN GONADS BY
SUPPRESSING GAMETOGENESIS
Unuma, T
Unuma, T. 2002. Gonadal growth and its relationship to aquaculture in sea urchins. In "TheSea Urchin: From Basic Biology to Aquaculture" ed. by Yokota, Y., Matranga, V.,
Smolenicka, Z., Swets & Zeitlinger, Lisse, pp.115-127.
The sea urchin gonad contains two main types of the cells: nutritive phagocytes (NPs)and germ cells (GCs). NPs store nutrients necessary for gametogenesis, such as proteins,carbohydrates and lipids. The most abundant protein in NPs is a glycoprotein called the
major yolk protein (MYP), originally identified as the most predominant component of yolk
granules in the eggs. The proportion of NPs and GCs varies with the maturation of the
gonads. Before gametogenesis, NPs fill the gonadal lumina and increase in size byaccumulating nutrients derived from food. As gametogenesis proceeds, NPs supply MYP
and other nutrients to the developing GCs and decrease in size. In fully mature gonads, anumber of ripe ova or spermatozoa fill the gonadal lumina, whereas NPs lose their nutrientsand shrink. The quality of the gonad as a food product usually deteriorates while GCs
proliferate and develop. Promotion of NP growth and suppression of gametogenesis are
prerequisites to achieving high profitability in sea urchin aquaculture.

Unuma, T., Walker, C.W. 2009. Relationship between gametogenesis and food quality in sea
urchin gonads. Proceedings of the 36th U.S.-Japan Aquaculture Panel Symposium. pp.45-54.
The sea urchin gonad contains two main types of the cells: germinal cells (GCs) and somatic
nutrient storage cells called nutritive phagocytes (NPs). The proportion of GCs and NPs varies during
the year. Before gametogenesis, NPs fill the gonadal lumina and increase in size by accumulating
nutrients derived from food. As gametogenesis proceeds, NPs decrease in size as nutrients aremobilized and transferred to GCs. In fully mature gonads, ova or spermatozoa fill the gonadal lumina
and NPs shrink to their smallest size. The best season for eating gonads is restricted to a few months
around the initiation of gametogenesis when NPs begin to mobilize their nutrients. Before that period,
the size of the gonad is too small. After that period, as gametogenesis proceeds, the quality of gonads
as food products gradually decreases. The tissues of the gonads become fragile as they mature. Afterthe ripe gonads are removed from the test, ova or spermatozoa ooze from the gonoduct, reducing
their commercial value. In some species, strong bitterness develops in the ovary as oogenesis
proceeds. To prolong the period during which commercially valuable sea urchin gonads can be
harvested and to improve the quality of the gonad, two strategies are proposed. One is to accelerate
the growth of NPs by feeding sea urchins with an artificial feed instead of macroalgae. The other is to9

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suppress gametogenesis by manipulating environmental cues that trigger gametogenesis or by
generating infertile, triploid sea urchins.

Walker, C.W., Lesser, M.P., Unuma, T. 2013. Sea urchin gametogenesis - structural,functional and molecular/genomic biology. In "Sea urchins: biology and ecology, 3rd edn."
ed. by Lawrence, J.M., Elsevier, Amsterdam, pp.25-38.
Gametogenesis and intra-gonadal nutrient storage and utilization are linked processes
during sea urchin reproduction. Dynamically interacting germinal and somatic cellular
populations make up the germinal epithelium of the sea urchin gonad. Uniquely among
edible animal reproductive products, sea urchin gonads grow in size not only because
gametogenesis increases the size and/or numbers of germinal cells present but also because
somatic cells within the germinal epithelium, the nutritive phagocytes, store extensive
nutrient reserves before gametogenesis begins. Here we provide an up-to-date discussion of
the unique nature of sea urchin gametogenesis including: gonad structure, the dual functionsof nutritive phagocytes, annual structural and molecular/genomic variation andenvironmental control. Knowledge of these phenomena has lead to successful manipulationof sea urchin reproduction and will increasingly provide opportunities for aquaculture
worldwide.

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MATURATION CONTROL OF THE SHORT-SPINED SEA URCHIN,STRONGYLOCENTROTUS INTERMEDIUS, BY LOW TEMPERATURE REARING
USING DEEP-SEA WATER, WITH THE AIM OF EXTENDING THE MARKETSEASON
Kayaba, T
Unuma T (2002) Gonadal growth and its relationship to aquaculture in sea urchins. In: Yokota et al (eds)
The sea urchin: from basic biology to aquaculture. Swets & Zeitlinger, Lisse, pp 115-127
The sea urchin gonad contains two main types of cells: nutritive phagocytes (NPs) and germ cells(GCs). NPs store nutrients necessary for gametogenesis, such as proteins, carbohydrates and lipids. The
most abundant protein in NPs is a glycoprotein called the major yolk protein (MYP), originally identified
as the most predominant component of yolk granules in the eggs. The proportion of NPs and GCs varies
with the maturation of the gonads. Before gametogenesis, NPs fill the gonadal lumina and increase in sizeby accumulating nutrients to the developing GCs and decrease in size. In fully mature gonads, a numberof type ova or spematozoa fill the gonadal lumina, whereas, NPs lose their nutrients and shrink. The
quality of the gonad as a food product usually deteriorates while GCs proliferate and develop. Promotionof NP growth and suppression of gametogenesis are prerequisites to achieving high profitability in sea
urchin aquaculture.

Kayaba T, Tsuji K, Hoshikawa H, Kikuchi Y, Kawabata K, Otaki I, Watanabe T (2012). Effect of low
temperature rearing, using deep-sea water, on gonadal maturation of the short-spined sea urchin,
Strongylocentrotus intermedius, in Rausu, Hokkaido. Fish Sci 78: 1263-1272
It has long been hoped that sea urchins could be served to visitors to seafood restaurants, hotels, andsummer festivals in Rausu, located in a world natural heritage site “Shiretoko,” during the summer tourist
season. However, to date this has not been feasible because of the sea urchin spawning season (July to
September), during which the quality of gonads, the edible part of sea urchins, decrease due to maturation.
In this study, we examined the possibility of suppressing gonadal maturation and maintaining high-quality
sea urchin gonads by low-temperature rearing using deep-sea water. Unripe sea urchins captured before
the spawning season were reared under two temperature conditions from June to September. In thosegroups reared at ambient temperatures (2.8–19.6°C), gametogenesis in both sexes progressed rapidly withincreased temperature, and almost all sea urchins reached full maturity by late July. In contrast, in groups
reared at low temperatures (2.1–5.1°C), gametogenesis progressed slowly and over 60% of the sea urchins
did not reach maturity even by early September. The feeding experiment also revealed that feeding withlive Saccharina diabolica could increase the gonadal volume efficiently under low-temperature. These
results demonstrate that low-temperature rearing, supplemented with feeding, is effective in suppressing
gametogenesis to allow for the harvesting of high-quality sea urchins during the summer tourist season.
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Bottger SA, Devin M, Walker CW (2006). Suspension of annual gametogenesis in North American green
sea urchins (Strongylocentrotus droebachiensis) experiencing invariant photoperiod – Applications forland-based aquaculture. Aquaculture 261: 1422-1431
Sea urchin fisheries are valuable commercial resources in the United States with processed gonads
sold in Japanese and American markets and maximum US salea of $150M US dollars in 1996. Wild
populations of sea urchins on all coasts of the US have been dramatically over-fished. Aquaculture of sea
urchins in land-based facilities can help restore commercial populations and preserve this ecologicallyimportant herbivore. In this study, we used invariant summer photoperiod to prevent gametogenesis in the
North American green sea urchins (Strongylocentrotus droebachiensis) maintained in a land-based
acuaclture system and provided a commercially available formulated feed that promotes maximum growthof intra-gonadal somatic nutrient storage cells called nutrient phagocytes. Results were compared withindividuals fed the same formulated feed under ambient photoperiod in cages in the ocean. Monthly
samples of the gonads from both treatments were evaluated for gonad index, volume fractions of cellularconstituents of the germinal epithelium, oocyte diameter and taste. Over the 5 months of this study, gonad
indices increased significantly (p < 0.001) in both treatments from 4.8% ± 0.9 (all values ± SE) initially to
20.5% ± 2.1 under invariant and 23.2% ± 1.4 under ambient photoperiod with no significant difference
between treatments (p = 0.55). Volume fractions of nutritive phagocytes increased to 80.3% ± 5.9 (initial
37.9% ± 7.1) in males and 71.0% ± 6.7 (initial 10.3% ± 4.0) in females (p < 0.001) only under invariant
photoperiod. Nutritive phagocyte lengths increased under both photoperiod treatments, but the volume
fraction containing nutrients was higher under invariant photoperiod. Volume fractions ofgonial/gametogenic cells increased significantly (p < 0.001) only under ambient photoperiod from 20.4%± 5.5 to 37.8% ± 1.8 in males and 0% to 22.6% ± 3.6 in females. The volume of fraction of residual
oocytes from last year’s oogenesis increased under invariant photoperiod while that of both residual andnew oocytes increased under ambient photoperiod. Residual oocyte diameters increase from 56.2 m ±2.2 initially to 93.5 m ± 3.7 under invariant and those of residual and new oocytes to 126.0 m ± 7.3
under ambient photoperiod. Invariant photoperiod yields gonads in both sexes of S. droebachiensis that do
not initiate fall gametogenesis but attain large size as their nutritive phagocytes grow substantially in size.
A Canadian study of wild-collected S. droebachiensis indicated that gonads taste best when they containpre-dominantly nutritive phagocytes and not copious gametes, however gonad taste in our study was
unsatisfactory suggesting that the only commercially available sea urchin diet requires modification to
support commercial development of land-based aquaculture.

Nabata S, Hoshikawa H, Sakai Y, Funaoka T, Oohori T, Imamura T (1999) Food value of several algae
for growth of the sea urchin, Strongylocentrotus nudus (in Japanese with English abstract). Sci Rep
Hokkaido Fish Exp Stn 54: 33-40
Rearing experiments of the sea urchin, Strongylocentrotus nudus, were carried out during June toJuly, 1995 and June to August, 1996, using as food marine algae which settled and grew on the coralline
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UJNR Annotated Bibliography

flats after the removal of sea urchins. Feeding rate and growth rate at 17°C of the sea urchin fed on
Laminaria, Undaria, Costaria were high and those for Sargassume, Polysiphonia, Dictyopteris,Desmarestia were low in the single food item experiment. Among the algae supplied as food, the daily
amount of food eaten was high un large-sized groups of the sea urchins, while the small-sized groups
show the highest feeding rate. To examine any effect on gonad growth, we fed 3 algae, Laminaria,
Sargassume and Polysiphonia to sea urchins. Two month later, the gonad index was found to be thehighest in the Laminaria fed group. Among the algae fed, based on the gonad growth, the highest feeding
rate and the highest growth rate, we estimated the Laminariales are the most nutritionally valuable algae
for growth of Strongylocentrotus nudus.

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UJNR Annotated Bibliography

RELATIONSHIP BETWEEN SPAT DENSITY, FOOD AVAILABILITY, AND
GROWTH OF SPAWNERS IN CULTURED JAPANESE SCALLOP(MIZUHOPECTEN YESSOENSIS) IN FUNKA BAY: CONCURRENCE WITH ENSO
Baba, K
Baba K, Miyazono A, Matsuyama K, Kohno S, Kubota S (2007). Occurrence anddetrimental effects of the bivalve-inhabiting hydroid Eutima japonica on juveniles of theJapanese scallop Mizuhopecten yessoensis in Funka Bay, Japan: relationship to juvenile
massive mortality in 2003. Marine Biology 151: 1977-1987
In November 2003, we first observed prevalent occurrence of a hydroid, Eutima
japonica, on soft body tissues of age zero Japanese scallop (Mizuhopecten yessoensis)
juveniles cultured in large areas of Funka Bay, Hokkaido. The occurrence coincided withmassive death of juvenile scallops. A major objective was to clarify ecological relationships
between the symbionts, and to infer the relationship between symbiosis and the massive
mortality. To do this, we investigated distributions of association rates of hydroids withjuvenile scallops at 15 to 34 sites over three years (2003-2005), with age one adult scallops
at 24 sites in 2003, and with mussels at 13 sites in 2004. We studied seasonal changes inassociation rates with juvenile scallops, and numbers of polyps per juvenile scallop at threesites from November 2003 to June 2004. We also quantified the hydroid impacts on juvenile
scallop shell length growth and triglyceride accumulation in the digestive gland. The
association rate of E. japonica polyps with juvenile scallops was high in large areas of Funka
Bay in 2003, and overlapped the distribution of mussels bearing polyps. Association rates
with age one adult scallops were very low in November 2003, even at the sites where polyps
were very common on juvenile scallops. Levels of hydroid occurrence in juvenile scallops
vary by year. We found that hydroids presence in juvenile scallops declined drastically in2004 and 2005. The association rates with juvenile scallops, and numbers of polyps per
juvenile scallop declined during winter, until they disappeared completely in the following
June. Since polyps were rare in adult scallops, we believe that infection of juvenile scallops
was probably initiated from the planulae produced by medusae released from polyps
growing on Mytilus spp., especially M. galloprovincialis. Subsequently, the inhabitation
spread intraspecifically and interspecifically. In juvenile scallops, inhabitation of polyps
reduced shell length growth by 43 %, and triglyceride accumulation in digestive glands by
24 to 47 %. Inhabitation of E. japonica on juvenile scallop is best regarded as parasitism,
rather than inquilinism or commensalism. Occurrence of polyps was probably not a direct
lethal factor for juvenile scallops, because there were some sites where association rates
were high, but mortalities were low. Massive mortalities in 2003 may have resulted fromsimultaneous impacts of heavy polyp load and stresses caused by the way in which theanimals were handled (transferred from cages for pre-intermediate culture to cages for
14

UJNR Annotated Bibliographyintermediate culture), because the massive mortality occurred within a month of the transfer.
The presence of polyps in juvenile scallops does not affect the quality of the product inFunka Bay, because market size scallops are hydroid-free.

Baba K, Miyazono A, Kohno S, Kubota S (2007). The symbiosis of the bivalve-inhabitinghydroid Eutima japonica in juvenile of Japanese scallop Mizuhopecten yessoensis in Funka
Bay: preventive measure against the symbiosis. Sci.Rep.HokkaidoFish.Exp.St 76: 45–49
To make preventive measures against symbiosis of the bivalve-inhabiting hydroid
Eutima japonica with cultured juvenile (age zero) scallop Mizuhopecten yessoensis, weinvestigated the ecological relationships between the hydroid, the main host Mytilus
galloprovincialis, and the juvenile scallop in Funka Bay, Hokkaido, Japan. Interannual
variation of the association rate with juvenile scallop was very large; the mean rates were
between 9.3 % (2004) and 69.5 % (2006). The period of the inhabitation in the juvenile
scallop was from September to February, and the spread and cessation of the inhabitation
occurred rapidly within about one and two months, respectively. The peak of medusa bud
was July in M. galloprovincialis, whereas the inhabitation occurred from beginning of
September in the juvenile scallop. The symbiosis probably initiates when the juvenile scallop
encounter with the planula larva. Since the hydroid does not inhabit in adult scallop nor age
zero M. galloprovincialis, high age M. galloprovincialis, especially inhabiting at the culture
facilities of the scallop, is probably the source of the planula larvae of the hydroid for the
inhabitation in the juvenile scallop. Therefore, the cleaning up the facilities after harvest of
the scallop is considered to be effective to prevent the inhabitation of the hydroid in thejuvenile scallop.
Baba K, Miyazono A, Hara A (2007). Relationship between vitellin concentration in
hemolymph and oocyte necroses during annual reproduction cycle in the Japanese scallop
Mizuhopecten yessoensis. Invertebrate Reproduction and Development 45: 175–184
A specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed
for measurement of vitellin in the hemolymph and ovary of the Japanese scallop(Mizuhopecten yessoensis). We observed two peaks of vitellin concentration in thehemolymph during the annual reproductive cycle. The peaks corresponded with theoccurrence of two types of oocyte necrosis, type-I and type-II. Type-I necrosis occurs in theearly phase of vitellogenesis and is characterized by necrotic oocytes that do not have agerminal vesicle. Type-II necrosis occurs during the mid- to late-phase of vitellogenesis andpeaks just before spawning. It is characterized by necrotic oocytes whose germinal vesicle isstrongly stained by eosin - i.e., the germinal vesicle has lost basophilic properties. In
immunohistochemical examinations, oocytes and auxiliary cells (which are in contact with15

UJNR Annotated Bibliography

oocytes), reacted with anti-vitellin serum; mid-gut gland, adductor muscles, intestine,
gonoduct, kidney, and gill do not. These results suggest that the presence of vitellin in thehemolymph is a result of leakage of yolk protein from necrotic oocytes.

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UJNR Annotated Bibliography

OYSTER CULTURE IN HOKKAIDO, JAPAN
Hasegawa, N
Inukai T, Nishio S (1937). A limnological study of Akkeshi Lake with special reference to the
propagation of the oyster. Journal of the Faculty of Agriculture, Hokkaido Imperial University 40: 1-33.
This paper published in 1930s when fisheries of natural oyster were declining in Akkeshi, Hokkaido,
Japan and reported that the greater part of this area where oysters were formerly produced is covered onlywith dead shells, and live oysters are found with great difficulty at that time. This paper also reported
research data about the environmental condition in this estuary and discuss the reconstructive procedurefor oyster fisheries as follows; Temperature of the surrounding medium is the primary importance, firstfor the general metabolism and, then, the reproductive activity of the oyster. In Akkeshi-ko estuary(Akkeshi Lake) the water temperature is always above 8˚C from the beginning of May to the end of
November in ordinary years and this is by no means short compared with the other oyster producing
localities. Unfortunately, the water in Akkeshi-ko rarely has a temperature higher than 25˚C and even if it
becomes warmer than 25˚C for short time. Thus it often happens that there are few and sometimes noseeds oysters in this Lake. The only economical method of cultivating oyster in the lake is secured bytransplanting the oyster seed from districts such as Miyagi.

Usuki H (2002). Evaluation of characteristics and preservation of Pacific oyster, Crassostrea gigas, inview of the genetic resources. Bulletin of Fisheries Research Agency 40: 40-104.
In this paper, differences in characteristics among local populations of Pacific oyster (Crassostrea
gigas) in Japan (Hiroshima, Miyagi, Ariake and Goto), evaluation of several characteristics in their
genetics and environments, estimation of heritability for improvement of economic traits, andpreservation of sperm and larvae were studied on the basis of genetic resources.
Particularly, in this studies concerning adults, the differences in characteristics among local
populations were reported as follows; The Miyagi population grew fastest and the Ariake and Goto
populations grew slower for both generations (parental and progeny groups cultured in Hiroshima Bay).
Though the mortality was not significantly different for progeny cultured in pocket mesh bags, theprogeny from Goto showed significantly higher mortality among progenies cultured by the collector
method. Progeny from Ariake showed significantly lower mortality among the other progenies when
cultured in narrow (10 mm) opening mesh bags and is thought to be the most tolerant to deterioration ofliving environment. Progeny of the Ariake population also had a spiny projection on their valves andshowed a higher biomass rate.

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UJNR Annotated Bibliography

Itoh N, Oda T, Ogawa K, Wakabayashi H (2002). Identification and Development of a Paramyxean
Ovarian Parasite in the Pacific Oyster Crassostrea gigas.

Fish Pathology 37: 23-28.

The intracellular ovarian parasite causing unaesthetic appearance in the ovary of Pacific oysters
Crassostrea gigas in Japan has remained unidentified since the first report of its occurrence in the
1930s.The parasite was observed through transmission electron microscopy, and identified as theparamyxean Marteilioides chungmuensis, which was first reported from Pacific oysters in Korea. In orderto obtain information on the development of the parasite, the diameters of both the parasite and theinfected ova were measured. The growth of the parasite was highly correlated with the growth andmaturation of host gonad cells. Histological observations suggest that M. chungmuensis invades immature
ova, which move to the center of the follicle along with its development, and infected ova are finallyreleased outside through the genital canal.

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UJNR Annotated Bibliography

MODELING INTRASPECIFIC GENETIC EFFECTS FOR MANAGEMENT OF
AQUACULTURE PROGRAMS
Volk, J
Jensen, Ø. T. Dempster, E. B. Thorstad, I. Uglem, A. Fredheim. 2010. Escapes of fishes from
Norwegian sea-cage aquaculture: causes, consequences and prevention. Aquaculture Environment
Interactions, 1: 71-83.
The authors broadly discuss the magnitude of the problem of escapes from salmon and cod cage-farming
aquaculture operations in Norway and provide specific recommendations to prevent escapes. Current
knowledge about the extent of threats presented by escapes in terms of economic and ecological impacts are
discussed in the context of experiences in the Norwegian aquaculture industry. While escapes occur due toseveral internal and external factors, reports from fish farming companies indicated that cage failure was by
far the most common cause of large-scale escape in Atlantic salmon farming operations. They report studies
that show the mechanisms of escape are not the same across species. Atlantic cod may cause more wear tonets and be more likely to escape through tears in the net. Consequences of escape, such as disease transfer,interbreeding, competition, and predation are generally discussed as areas for further research. The main
message of this paper is that prevention is the best tool to reduce the risk of escapes. The authors report
evidence that the level of escapes from cages was greatly reduced in Norway following legislation that has
specific requirements for design of farms and the handling and use of equipment. They recommend countries
develop similar measures to reduce escapes such as mandatory reporting of escapes, a process to use thesereports to develop better standards, mandatory technical assessments following reported “large” escape
events, technical standards for equipment, and finally, identification of key operational components that havea higher potential to cause an escape event, including training of staff to reduce human errors. This paper
was used to identify mechanisms of escape through cage failure and operational scenarios, which are central
to development of the escapes component of the OMEGA simulation model.

Meager, J. J., J. E. Sklæraasen, A. Fernö, and S. Løkkeborg. 2010. Reproductive interactions between
fugitive farmed and wild Atlantic cod (Gadus morhua) in the field. Canadian Journal of Fisheries andAquatic Sciences, 67:1221-1231.
The authors present a study of spawning interactions between cultured and wild Atlantic cod andtested the potential for hybridization between farm escapes and wild conspecifics. Using a spatial andtemporal analysis of wild and farmed cod tracked through biotelemetry, positioning of fish based on sex
and origin indicated that farmed fish behave differently from wild fish relative to spawning groundlocation. However, despite these differences, hybridization is likely, especially between farmed females
and wild males. The results illustrate that behavioral differences between cultured and wild fish may notpreclude spawning interactions. The authors conclude there is a high potential for farmed cod to hybridize
with wild fish. They recommend further research should be a priority to further understand theconsequences of interbreeding and to identify methods for escape prevention.
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UJNR Annotated Bibliography

Ford, M. J. 2002. Selection in captivity during supportive breeding may reduce fitness in the wild.
Conservation Biology 16:815–825.
Much of the concern surrounding effects of escaped cultured fish involves interbreeding with wild
conspecifics and potential loss of genetic fitness of the wild population. Ford presents a single trait
phenotypic model that assumes different optimum trait values for the culture and natural environments.
The Ford model describes how mean phenotype values of captive and wild fish shift relative to optimum
values for the environments based on gene flow between escapees (or captive breeding) and wild fish. The
results suggest that cultured fish can have a strong influence on the fitness and sustainability of wild
populations depending on the amount of interbreeding. The level of effect depends on the details of themodel such as differences in optimum trait value, selection pressure, and trait heritability. Controllinggene flow between wild and cultured fish can potentially reduce the domestication effect in wild
populations. Overall outcomes of fitness in the wild also depend heavily on habitat capacity andpopulation dynamics. This model has been used in several other studies to explore the potential
consequences to wild population fitness from captive breeding to increase the size of wild populations,and from the unintended straying of cultured fish to wild populations.

Paquet, P.J., T. Flagg, A. Appleby, J. Barr, L. Blankenship, D. Campton, M. Delarm, T. Evelyn, D.
Fast, J. Gislason, P. Kline, D. Maynard, L. Mobrand, G. Nandor, P. Seidel & S. Smith (2011):
Hatcheries, Conservation, and Sustainable Fisheries—Achieving Multiple Goals: Results of the
Hatchery Scientific Review Group's Columbia River Basin Review, Fisheries, 36:11, 547-561
The Hatchery Scientific Review Group (HSRG) was established by the U.S. Congress to review
salmon and trout hatchery programs in the Pacific Northwest with the goal of recommending hatcheryreform guidelines while still retaining the goal of providing fish for harvest and conservation goals fornatural populations. This paper presents the approach used and recommendations that included anassessment of 178 hatchery programs and 351 salmonid populations within the Columbia River Basin.
This approach included a scientific framework and three principles to guide their assessment: 1) “clear
and specific quantifiable goals for harvest and conservation,” 2) “be scientifically defensible,” and 3)“include monitoring and evaluation of benefits and risks.” HSRG used the All-H Analyzer (AHA) model
to evaluate dynamics of populations in the Columbia River system through an integrative analysis ofseveral factors related to hatchery operations, and productivity and capacity of wild populations. Using
model simulation results, HSRG formulated a working hypothesis for baseline conditions and to evaluate
strategies to better achieve stated goals for hatcheries and wild populations. The HSRG concluded thatthrough following recommended management and harvest practices, such as broodstock selection andselective harvest, hatcheries can serve dual goals of contributing to harvest while remaining compatible
with or contributing to conservation goals. The guiding principles stated by the HSRG are the core
purpose of the OMEGA model.

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UJNR Annotated Bibliography

FLUOROCHROME MARKING OF OUT-PLANTED GREEN SEA URCHINS,STRONGYLOCENTROTUS DROEBACHIENSIS, FOR SEA RANCHING ANDRESTOCKING PROGRAMS IN THE GULF OF MAINE, USA
Eddy, S
Dumont, Clément, John H. Himmelman, and Michael P. Russell. 2004. Size-specific movement ofgreen sea urchins Strongylocentrotus droebachiensis on urchin barrens in Eastern Canada. Marine
Ecology Progress Series 276:93-101.
The authors were the first to use fluorescent tagging as a tool to follow the movement of green sea
urchins in the field. They tagged all urchins within 1m2 plots with calcein fluorochrome and quantified
their abundance 9 to 40 days later in the release plots and at different distances away from the release. Theysaw that movement was size dependant, with urchins > 15 mm diameter more likely to move into or out ofthe plot areas. This movement was most likely in response to food availability. Urchins < 15 mm weremore sedentary and therefore were more likely to be found within the release area. These movement
patterns will likely result in size partitioning of sea urchins released for out-planting.

Ellers, O. and A.S. Johnson. 2009. Polyfluorochrome marking slows growth only during themarking month in the green sea urchin Strongylocentrotus droebachiensis. Invertebrate Biology 128:126-144.
The authors demonstrate the use of polyfluorochrome marks, whereby urchins are sequentially taggedwith different fluorochromes to permit unique identification of individuals or cohorts. They used alizarin
compexone, calcein, calcein blue, and tetracycline as markers, administered by immersion with small green
sea urchins (S. droebachiensis) and injection with larger urchins. Multiple or sequential marking can beused to measure multiple growth points. They showed that there was only a transient affect on juvenile
growth in the first month following marking but no long term effects on growth or survival. They
quantified growth using a nominal diameter, or fitted constant, times the cube root of weight. This
increased the precision of measurements by a factor of 6 relative to measured diameter. The authors showthat growth rate was a decreasing function of diameter except for a growth lag in the smallest urchins.

Kirchhoff, N., Eddy, S., Harris, L. and N. Brown. 2008. Nursery-phase culture of green sea urchin
Strongylocentrotus droebachiensis using "on-bottom" cages. Journal of
Shellfish Research. 27(4):921-927.
This study was the first in the Gulf of Maine to show differences between ocean lease sites used as
nurseries for green sea urchins. Two adjacent lease areas with differing bottom substrates, known as Job
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UJNR Annotated Bibliography

and Sloop, were compared as nursery areas for juvenile growth of green sea urchins S.
droebachiensis. Juvenile urchins (avg. 7.9 ±0.7 mm diameter) were captured from the lease areas,measured, and placed into mesh tubes attached to on-bottom oyster cages. A small amount of kelp wasplaced in each tube at the start, but no additional feed was added during the ensuing 6 months. Groups ofurchins were counted and measured 5 weeks, 3 months, and 6 months after placement to gauge growth, andsurvival. Final survival indicated that cobble bottom type supported the highest survival at both Job Island(89%) and Sloop Island (71%), followed by Sloop mussel (59%) and Sloop ledge (56%). After 6 months
the average diameter reached 11.08 +/- 1.49 mm. Final average test diameter was significantly larger at
Sloop ledge (12.17 mm) and Sloop mussel (12.58 mm), than at Sloop cobble (9.83 mm) and Job cobble(9.66 mm). These results suggest on-bottom culture through the critical nursery phase is technicallyfeasible and may represent an economical way to rear hatchery produced green sea urchin seed to the‘planting out’ size.

Johnson, A.S., Salyers, J.M. Alcorn, N.J., Ellers, O., and J.D. Allen. 2013. Externally visible
fluorochrome marks and allometrics of growing sea urchins. Invertebrate Biology, Early View - Wiley
Online Library DOI: 10.1111/ivb.12024: 1-19.
The authors marked 2 month old S. droebachiensis with the fluorochromes calcein, calcein-blue andtetracycline by batch marking with immersion. They show for the first time that fluorochrome tags can be
externally visualized for up to 342 days on the skeletal plates, demipyramids, or spines in 100% of green sea
urchins marked with calcein, and in 22% marked with tetracycline. Marks were brightest withcalcein-marked and faintest on tetracycline-marked sea urchins, in correspondence with the dose. In a
separate experiment they show that multiple marks of the above fluorochromes plus alizarin complexone,
administered by injection to larger urchins, can persist internally for at least 2 years. The ability toexternally visualize fluorochrome marks in sea urchins enables multiple recapture and release studies of thesame groups or even individuals in field and laboratory studies. This also make is possible to track the fateof hatchery-reared sea urchins released on-bottom with non-lethal sampling for at least 11 months andperhaps longer. The development of portable instruments that can be used in the field to visualize external
fluorescence seems possible, and will be a powerful tool for research and aquaculture.

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UJNR Annotated Bibliography

SPECIFIC MONOVALENT AND DIVALENT ION SUPPLEMENTATION FOR
CULTURE OF MARINE SPECIESGreen, C
Fisher, C., C. Bodinier, A. Kuhl, and C. Green. 2013. Effects of potassium ion supplementation on
survival and ion regulation in Gulf killifish Fundulus grandis larvae reared in ion deficient saline
waters. Comparative Biochemistry and Physiology, Part A. 164:572-578.
This study encompasses results pertaining to the K+ ion manipulation portions of the abstract presented
above for the UJNR Scientific Symposium. This investigation increased [K+] from values similar to
freshwater sources (0.33 mM) to concentrations of 2.96 mM, found saline waters at 10‰. A number ofbiochemical and molecular techniques were performed to examine the effect of this K + ion gradient, which
included: whole body ion composition, Na+/K+ -ATPase (NKA) activity, gill ionocyte morphometrics,
relative gene expression (NKA, NKCC, and CFTR), and corresponding immunocytochemistry at the gilland intestinal epithelium. Results more tangible to aquaculturist, such as growth and survival, indicated thepresence of a threshold within the gradient examined whereby survival was not different between
treatments of 1.3 to 2.9 mM [K+]. Utilizing immunocytochemistry, the differences between these
seemingly similar treatment groups indicated that the treatments between 1.3 to 2.9 mM [K +] were different
in terms of gill ionocyte area and NKA/CTFR localization.

Ostrowski, A.D., Watanabe, W.O., Montgomery, F.P., Rezek, T.C., Shafer, T.H., Morris Jr, J.A., 2011.
Effects of salinity and temperature on the growth, survival, whole body osmolality, and expression of
Na+/K+ ATPase mRNA in red porgy (Pagrus pagrus) larvae. Aquaculture 314, 193-201.
These authors investigated a number of physiological parameters pertaining to salinity andtemperature in embryos and larvae of red porgy (Pagrus pargrus), which is viewed as a high-market valuemarine finfish with good potential as an aquaculture species. Embryos and resulting larvae were reared atfour temperatures (17, 19, 21, 23 °C) and two salinities (24 and 34‰). Larvae (16 dph) were transferredfrom their respective salinities to 44‰ to represent a sublethal hyperosmotic challenge. These authors
demonstrated significant increases in NKA mRNA expression in individuals acclimated to 24‰ at 24 hafter transferred to increased salinity, while individuals from 34‰ exhibited no significant changes in NKA
expression. Temperature was not observed to influence expression of NKA, while metabolic parameters
related to growth were influenced by the temperature gradient in their study. The authors utilized
traditional growth metrics including molecular tools to anticipate optimum salinity (24‰) and temperature(23°C) conditions that are optimum for larval rearing of this species.

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Bodinier, C., Sucré, E., Lecurieux-Belfond, L., Blondeau-Bidet, E., Charmantier, G., 2010. Ontogenyof osmoregulation and salinity tolerance in the gilthead sea bream Sparus aurata. Comparative
Biochemistry and Physiology - Part A: Molecular and Integrative Physiology 157, 220-228.
The gilthead sea bream (Sparus aurata) is a commercially important aquaculture species, which
spawns in the open ocean and whose resulting larvae and juveniles migrate to lower salinity estuaries andlagoons. These authors investigated the development of salinity tolerance in gilthead sea bream from 3, 30,
75, 96, and 300 d post hatch (dph) by challenging them with 9 salinities ranging between freshwater and45.1‰. Utilizing immunohistochemistry, these authors localized the NKA throughout these challenges todocument location of ion regulation within respect to this ionoregulatory protein. Initially, immunopositive
NKA ionocytes were located in the integument along the yolk sac and integumentary folds representing thebranchial slits. A functional shift from integument to gills was demonstrated 30 and 70 dph, when both the
integument and gills were observed to locally express NKA, whereby from 70 to 300 dph the gills remain
the main site of osmoregulation. Increases in osmoregulatory capacity for this species at the intervals
examined within this study related to the shifts and patterns observed through immunohistochemistry.

Christensen, A.K., Hiroi, J., Schultz, E.T., McCormick, S.D., 2012. Branchial ionocyte organizationand ion-transport protein expression in juvenile alewives acclimated to freshwater or seawater. The
Journal of Experimental Biology 215, 642-652.
In gill ionocytes, ion transport is activated by the basolaterally located NKA which generates anelectrochemical gradient by coupling two extracellular K+ with three intracellular Na+, driving the ionsaccording to expression, location, and abundance of other proteins such as NKCC and CFTR. Christensen
et al. investigated changes in alewife physiology and branchial epithelium as individuals were acclimated to
freshwater or saltwater. This represents the first study of its kind to characterize multiple ion-transport
proteins in a non-salmonid anadromous fish. Corresponding increases in NKA, NKCC1, and CFTR
abundance at the gill epithelium with increasing salinity was used to establish a gill model for
hypo-osmoregulation. In gill ionocytes, NKA is responsible for lowering intracellular Na + allowing thebasolateral NKCC1 to import Na+, K+, and two Cl- ions. Excess intracellular Cl- is then secreted through the
CFTR chloride channel. NKCC1, the secretory isoform is expressed basolaterally in the gill ionocytes,
while NKCC2 is identified as an absorptive isoform and is expressed apically along the intestinal andurinary bladder epithelium of saltwater and euryhaline teleosts. The authors determined the key
differences between freshwater and seawater acclimated alewives in the context of ion transporters at thegill epithelium. The implications of these investigation has assisted in increasing the information on
multicellular complexes of mature ionocytes and the role of salinity and specific ion in the maintenance of
homeostasis.

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UJNR Annotated Bibliography

GOOD AQUACULTURE PRACTICES TO REDUCE THE USE OF
CHEMOTHERAPEUTIC AGENTS, MINIMIZE BACTERIAL RESISTANCE, AND
CONTROL PRODUCT QUALITY
Serfling, S
Karunasagar, Iddya 2012. International trade, antimicrobial agents, and antibiotic resistance: Publichealth and trade impact of antimicrobial use in aquaculture. FAO Fisheries and Aquaculture
Technical Paper No. 547, Pages 1-9.
The importance of antimicrobial agents in protection of animals has been widely acknowledged, but
the negative impact and the use of these agents in animals raised for food have been a cause ofconcern. The Food and Agriculture Organization of the United Nations (FAO), the World HealthOrganization (WHO) and the World Organization for Animal Health (OIE) have organized several expert
consultations and technical meetings to review the global situation and develop recommendations.
The author outlines how detection of certain banned antibiotics in fish and crustaceans in international
trade during 2001-2002 lead to greater attention on the public health risks owing to the use of antimicrobials
agents in aquaculture. Most fish importing countries adopt a zero tolerance approach regarding residues of
antimicrobials that are banned for use in food animals. In such cases, residue levels that attract regulatory
action are based on analytical capability rather than toxicology of the residues. Development and spread ofantibiotic resistance has been a cause of concern, although this issue is complicated by possible multiple
origins of resistance traits found in aquatic bacteria.

Brett Koonse 2012. Good aquaculture practices to minimize bacterial resistance. FAO Fisheries andAquaculture Technical Paper No. 547, Pages 25-28.
The author identifies how the preventive measures associated with Good Aquaculture Practices can beapplied to hatcheries and farms, by government regulators, and by academics and health careproviders. There are a number of physical, chemical and biological precautionary measures that can beundertaken.


Physical barriers are intended to prevent disease carrying vectors from entering a hatchery or farm site,and may include physical barriers, water treatment and quarantine. Physical barriers may also be proper
location of the farm, stress reducing practices like not over-crowding in pens or ponds; avoiding improper
temperatures, consistent dissolved oxygen excessive handling and improper diet.
Chemical controls include those that are sued to prevent the introduction of pathogens or vectors bytreating materials before they enter the facility. For example chlorination or ozone can be used to treat
incoming water, and iodine and chlorine can be used to treat other potential vectors such as footwear andclothing.

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UJNR Annotated BibliographyBiological controls include those measures that prevent or treat infections and the proper use of

chemotherapeutic agents or the use of vaccines. It may also include the use of specific pathogen-free
shrimp and management practices to prevent bacterial contamination of aquaculture products, grow-out
ponds or cages. This includes having proper toilet facilities, sanitary waste removal, healthy workers,
effluent treatment and proper security at the site.

Michael L. Jahncke, Craig L. Browdy, Michael H. Schwarz, Al Segars, Juan L. Silva, David C. Smith,
Alvin D. Stokes, 2002. Risk management, disease prevention, and HACCP principles: Applicationof Hazard Analysis Critical Control Point (HACCP) Principles as a Risk Management Tool to Control
Viral Pathogens at Shrimp Aquaculture Facilities. NOAA Office of Sea Grant, Virginia Graduate
Marine Science Consortium and the Virginia Sea Grant College Program, Publication VSG-02-10,
Pages 1-33.
The authors identify how the principles of HACCP (Hazard Analysis Critical Control Point) can beused as a preventive risk management system to control the introduction of pathogens and the outbreak of
disease in shrimp aquaculture facilities. The HACCP approach requires a hazard analysis that identifies a
potential hazard in the system and then a Critical Limit with a maximum and or minimum point is set foreach component of the system. When monitoring the aquaculture system and a critical limit has been in
violation, then a corrective action is taken to bring the system back into compliance.
To develop an effective biosecurity program fish and shrimp farmers should follow these principles
base on seafood HACCP: 1) Perform Systematic Hazard Analysis, 2) Determine Critical Control Points, 3)
Establish Critical Limits, 4) Determine Appropriate Corrective Actions, 5) Establish Monitoring
Procedures, 6) Establish Verification Procedures, 7) Establish Record Keeping Systems.

Brett Koonse, William Burkhardt, Stuart Chirtel, and George Hoskins, 2005. Food safety andaquaculture, controlling risk, coliform bacteria: Salmonella and Sanitary Quality of Aquaculture
Shrimp. Journal of Food Protection, Vol. 68, No 12, Pages 2527-2532.
The authors examined the prevalence of Salmonella and coliform bacteria on shrimp aquaculture farms
in several Asian countries to develop guidelines or preventative measures for reducing Salmonella and fecal
contamination on products harvested from these farms.
The U.S. Food and Drug Administration, in conjunction with foreign government regulatory agencies,
the aquaculture industry, and academia affiliates, analyzed 1,234 samples from 103 shrimp aquaculture
farms representing six countries between July 2001 and June 2003 for fecal coliforms, Escherichia coli, andSalmonella. A significant relationship was found between the log number of fecal bacteria and theprobability that any given sample would contain Salmonella. The likelihood of any given sample
containing Salmonella was increased by 1.2 times with each 10-fold increase in either fecal coliform or E.
coli concentration. The likelihood of finding Salmonella in grow-out pond water increased 2.7 times with26

UJNR Annotated Bibliographyeach log unit increase in fecal coliform concentration and 3.0 times with each log unit increase in E. coli
concentration. Salmonella is not part of the natural flora of the shrimp culture environment nor is it
inherently present in shrimp grow-out ponds. The occurrence of Salmonella bacteria in shrimp fromaquaculture operations is related to the concentration of fecal bacteria in the source and grow-out pond
water.

Andreu Rico, Tran Minh Phu, Kriengkrai Sataportnvanit, Jian Min, A.M. Shahabuddin, Patrick J.G.
Henriksson, Francis J. Murray, David Little, Anders Dalsgaard, Paul J. Van den Brink,2013. Antimicrobial use, Asian aquaculture, and international trade: Use of veterinary medicines,
feed, additives, and probiotics in four major internationally traded aquaculture species farmed in
Asia. Journal of Aquaculture, 10.1016, Pages 1-38.
The authors provided detailed information on the use of antimicrobials, parasiticides, feed additives
and probiotics used in Asian aquaculture to improve the health of the organisms and to prevent or treat
disease. The study reports the outcomes of a survey of chemical and biological products in 252 grow-out
farms and 56 farm supply shops in Bangladesh, China, Thailand, and Vietnam. The survey was conducted
in 2011 and 2012 and included the following farm groups: Penaeid shrimp, freshwater prawns, tilapia, andpangasius catfish farms.
Results were analyzed with regard to the frequencies of use of active ingredients and chemical classes,
reported dosages, and calculated applied mass relative to production. A range of farm management andfarm characteristics were used as independent variables to explain observed chemical use patterns reported
by farmers in each group. The use of veterinary agents was found to be significantly higher in Vietnamese
Pangasius farms.
Semi-intensive and intensive shrimp farms in China, Thailand, and Vietnam showed a decrease in theuse of antibiotic treatments. These farm groups utilized the largest amount of chemicals relative to
production, with feed additives and plant extracts, probiotics, and disinfectants, being the most usedchemical classes, mainly for disease prevention. The surveyed farmers generally did not exceed recommend
dosages of veterinary medicines, and nationally or internationally banned compounds.

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UJNR Annotated Bibliography

THE IMPORTANCE OF TAURINE AND N-3 FATTY ACIDS IN COBIANUTRITION
Watson, A
Watson, A.M., Barrows, F.T., Place, A.R. 2013. Taurine Supplementation of Plant Derived Protein and
n-3 Fatty Acids are Critical for Optimal Growth and Development of Cobia, Rachycentron canadum.
Lipids DOI: 10.1007/s11745-013-3814-2.
The authors describe the effect of alternative lipid sources as potential fish oil replacements for cobia,
Rachycentron canadum. The fish oil replacements were a thraustochytrid meal (TM + SOY) plus soybean
oil (~9% CL) or a canola oil supplemented with the essential fatty acids (CO+EFA) docosahexaenoic acid
(DHA) and arachidonic acid (ARA) (~8% CL). The TM+SOY diet performed equivalently to the fish oil
diet; both resulting in significantly higher growth rates, lower feed conversion ratios, and higher survival
than the CO+EFA diet. The poor performance of the canola oil diet was attributed to insufficient addition of
EFA in the supplemented canola oil source. Increasing levels of EFA in the supplemented canola oil above
0.5 g EFA kg-1 would likely improve results with cobia. A finishing period was also utilized following the
growth trial to determine if fillet fatty acid profiles would change to a more fish oil type profile. When fish
fed either of the fish oil replacement diets were switched to the fish oil control diet, fatty acid profiles of thefillets were observed to transition toward that of the fish oil diet and could be predicted based on a standard
dilution model.

Watson, A.M., Barrows, F.T., Place, A.R. 2013. Taurine Supplemented Plant Protein Based Diets with
Alternative Lipid Sources for Juvenile Gilthead Sea Bream, Sparus aurata. Journal of Fisheries andAquaculture 4:1 59-66.
The authors describe the effects of two alternative lipid sources as fish oil replacements for giltheadsea bream, Sparus aurata. The fish oil replacements were a thraustochytrid meal (TM + SOY) plus soybean
oil (~9% CL) or a canola oil supplemented with the essential fatty acids (CO+EFA) docosahexaenoic acid
(DHA) and arachidonic acid (ARA) (~8% CL). Sea bream is one of the most cultured species in Europe andits popularity is increasing in the United States. Finding adequate alternative lipid sources in addition tocomplete fishmeal replacement would allow the industry to continue to expand. Both the CO+EFA dietperformed equivalently to a fish oil based control diet while the TM+SOY diet resulted in significantly
increased growth rates. The authors concluded that both of these lipid sources were viable candidates forfish oil replacements in feeds for this species as long as taurine is adequately supplemented.

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UJNR Annotated Bibliography

Ueki, I., Roman, H.B., Hirschberger, L.L., Junior, C. Stipanuk, M.H. 2013. Extrahepatic tissues
compensate for loss of hepatic taurine synthesis in mice with liver-specific knockout of cysteine
dioxygenase. American Journal of Physiology: Endocrinology and Metabolism. 302:E1292-1299.
DOI: 10.1152/ajpendo.00589.2011.
The authors examined the effects of knocking out liver specific expression of cysteine dioxygenase(CDO), a key enzyme in the synthesis of taurine. Although this work was done in the mouse model, theparallels for the taurine synthesis pathway of vertebrates makes this work important for comparisons totaurine synthesis in teleost species. CDO expression increased in kidney, brown fat, and gonadal fat tocompensate for the loss of synthesis capacity in the liver. These tissues also had higher concentrations ofhypotaurine, which is consistent with the CDO pathway of taurine synthesis. The authors concluded that
this redundant capacity to maintain taurine synthesis indicates not only a response to changing dietarytaurine levels (mice were fed taurine deplete feeds) but underscores the importance of taurine to overallvertebrate physiology.

Kozlowski, D.J., Chen, Z., Zhuang, L., Fei, Y.J., Navarre, S. Ganapathy, V. 2008. Molecularcharacterization and expression pattern of taurine transporter in zebrafish during embryogenesis. Life
Sciences. 82: 1004-1011.
The authors cloned and functionally characterized the taurine transporter, a membrane boundtransporter. The 625 amino acid encoded for by the zebrafish TauT cDNA is highly homologous to
mammalian TauT. The authors were able to express the zebrafish TauT in a mammalian cell line andcharacterized it as a Na/Cl dependent transporter. The presence of the transporter was observed in the retina,
brain, heart, kidney and blood vessels of developing zebrafish embryos. Taurine and the TauT mRNA wereduring early cleavage stages, indicating a maternal input. This study provides clear evidence of theimportance of taurine in vertebrate development as knockout of TauT with antisense morpholino
oligonucleotides resulted in cell death. The taurine transporter may be critical in teleost development andregular maintenance of the circulating and whole body taurine pools.

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UJNR Annotated Bibliography

CHOLECYSTOKININ AND TRYPSIN RESPONSES OF LARVAL RED DRUM(SCIAENOPS OCELLATUS) IN RESPONSE TO ALGAE, LIVE PREY, AND INERTPARTICLES
Webb, K
Dockray, G.J. 2012.

Cholecystokinin.

Current Opinion in Endocrinology, Diabetes & Obesity,

Vol: 19:1. Pp 8–12.
The author provides an excellent review of the current understanding of the role of Cholecystokinin(CCK) in satiety and to a lesser extent, digestion.

The author focuses on the role of CCK in the cephalicphase and reviews current knowledge of both CCK activators and targets.

The author also discussed therole of Leptin in potentiating the effect on CCK on vagal afferent neurons. Of particular note in thismanuscript, the author mentions the role of GPR40 and long-chain fatty acids in the secretion of fatty
acids while much of the other literature focuses on protein hydrolysate / amino acid roles in promoting
CCK secretion.

Liou, A.P., D. I. Chavez, E. Espero, S. Hao, S.A. Wank, and H.E. Raybould. 2010. Proteinhydrolysate-induced cholecystokinin secretion from enteroendocrine cells is indirectly mediated by
the intestinal oligopeptide transporter PepT 1.

Am J Physiol Gastrointest Liver Phsyiol, Vol: 300:G.

Pp G895-G902.
The authors investigate the role of the PepT1 transporter as the direct mediator of Cholecystokinin(CCK) secretion in response to protein hydrolysate. Before this work, PepT 1 was considered a likely
mediator of direct mediation of CCK secretion due to previous work that showed a synthetic dipeptide,
Gly-Sar, used in PepT1 kinetic studies caused a dose-dependent inhibition of gastric motility consonantwith CCK secretion.

The work demonstrated that while Gly-Sar did inhibit gastric motility it had noeffect on eliciting CCK secretion from CCK-eGFP cells.

Based on this work, the authors concluded that

protein detection by intestinal I cells likely includes both direct and indirect detection but that PepT 1 does
not have a direct role in CCK secretion. The authors do however suggest that PepT1 may function
indirectly by stimulating a diazepam-binding inhibitor.

Cudenneca, B.,

M. Fouchereau-Peron, F. Ferry, E. Duclos, and R. Ravallec.

2012.

In vitro andin vivo evidence for a satiating effect of fish protein hydrolysate obtained from blue whiting(Micromesistius poutassou) muscle.

J Func Foods, Vol: 4.

Pp 271-277.

The authors in this work examine the role of protein hydrolysate from blue whiting on CCK
secretion in STC-1 cells as well as on long and short-term food intake of rats fed blue whiting muscle
30

UJNR Annotated Bibliographyhydrolysate (BWMH). The authors found that in the in vitro studies on STC-1 cells, peptides from the
BWMH stimulated CCK secretion in a dose-dependent response similar to that seen from other types ofprotein hydrolysate. In the in vivo study, the authors found that BWMH produced short-term reductions
in food intake but that this reduction was not reflected in the long term. The in vivo study did show thatblood CCK and GLP-1 levels were more than doubled in rats fed 100 and 250mg of BWMH following a
24h fast lending credence that these may be involved in the short-term decrease in consumption.

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UJNR Annotated Bibliography

REPRODUCTIVE DYSFUNCTION IN CULTURED SABLEFISH (ANOPLOPOMAFIMBRIA)
Guzman, J
Guzmán, J.M., Luckenbach, J.A., Swanson, P. 2013. Molecular characterization and quantification ofsablefish (Anoplopoma fimbria) gonadotropin and their receptors: Reproductive dysfunction infemale captive broodstock. General and Comparative Endocrinology, Vol. 193. Pp 37-47.
This study describes for the first time the cloning and characterization of gonadotropin subunits (fshb,
lhb and cga) and gonadotropin receptors (fshr and lhcgr) in sablefish. Interestingly, sablefish
gonadotropin subunits display some features never observed in vertebrates, such as two additional Cys
residues in the N-terminal region of Fshb and a lack of potential N-glycosylation sites in Fshb and Lhb
subunits. Wild female sablefish captured in the fall completed gametogenesis and spawned in captivity,and had elevated gonadotropin signaling (pituitary and ovarian gene expression and plasma steroid levels)
compared to those captured in summer, which did not mature. These non-maturing females indeed had
similar reproductive endocrine profiles as prepubertal F1 females, indicating a failure of the
gonadotropin-ovary axis. These data suggest that the onset of vitellogenic growth in sablefish is a periodthat is sensitive to environmental conditions, and a major point of impairment of reproduction in both wildand F1 fish maintained in captivity.

Smith, E.K., Guzmán , J.M., Luckenbach, J.A. 2013. Molecular cloning, characterization, andsexually dimorphic expression of five major sex differentiation-related genes in a Scorpaeniform fish,
Anoplopoma fimbria. Comparative Biochemistry and Physiology Part B: Biochemistry andMolecular Biology, Vol. 165. Pp 125-137.
Five key sex differentiated-related genes (foxl2, cyp19a1a, dmrt1, sox9a and amh) were cloned andcharacterized in sablefish, and their expression analyzed during early gonad development. These fivegenes were differentially regulated in developing ovaries and testes in sablefish; specifically, foxl2 andcyp19a1a seem reliable as molecular markers of ovarian differentiation, and dmrt1 and amh for testiculardifferentiation in sablefish. The assays developed in this study provide a foundation for ongoing researchon sex differentiation and developing strategies for sex control in this species.

Vidal, B., Pasqualini, C., Le Belle, N, Holland, C.H., Sbaihi, M., Vernier, P., Zohar, Y., Dufour, S.
2004. Dopamine inhibits luteinizing hormone system and release in the juvenile European eel: aneuroendocrine lock for the onset of puberty. Biology of Reproduction, Vol. 71. Pp. 1491-1500.
In various adult teleost fishes, Lh ovulatory peak is under a dual neurohormonal control that is32

UJNR Annotated Bibliography

stimulatory by gonadotropin-releasing hormone (GnRH) and inhibitory by dopamine (DA).In this study it
was investigated whether DA could also be involved in the inhibitory control of Lh at earlier steps ofgametogenesis by studying the model of the European eel, Anguilla anguilla, which remains at a
prepubertal stage until the oceanic reproductive migration. According to a protocol previously developedin the striped bass, eels received sustained treatments with GnRH agonist (GnRHa), DA-receptor
antagonist (pimozide), and testosterone (T) either alone or in combination. Only the triple treatment withT, GnRHa, and pimozide could trigger dramatic increases in Lh synthesis and release as well as in plasma
vitellogenin levels and a stimulation of ovarian vitellogenesis. Thus, in the prepubertal eel, removal of DA
inhibition is required for triggering GnRH-stimulated Lh synthesis and release as well as ovariandevelopment. This is the first demonstration of a pivotal role for DA in the control of Lh and puberty in ajuvenile teleost.

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DEVELOPMENT AND CHARACTERIZATION OF SEVERAL OPEN FORMULA
REFERENCE DIETS FOR MARINE FISH LARVAE
Rust, M
Barrows, F. T. and W. A. Lellis, 2000.
Aquaculture.

Microbound feeds.

John Wiley & Sons, New York.

In R. R. Stickney (ed) Encyclopedia of

525-528.

In addition to a review of methods to make older micro bound feeds for aquaculture, methods to
produce microparticulate feeds using micro extrusion and particle assisted rotational agglomeration are
explained.

Cho, C. Y., C. B. Cowey and T. Watanabe. 1985. Finfish Nutrition in Asia: Methodologicalapproaches to research and development.

IDRC, Ottawa, Ont. Canada.

154 pages.

Step by step instructions to produce zein and carrageenan bound diets developed in Japan are
explained in English.

Holt, G. J. (editor) 2010.

Larval Fish Nutrition.

John Wiley & Sons, New York. 435 pages

Nutrition is particularly important in the healthy development of fish during their early-life stages.
Understanding the unique nutritional needs of larval fish can improve the efficiency and quality of fish
reared in a culture setting. Larval Fish Nutrition comprehensively explores the nutritional requirements,
developmental physiology, and feeding and weaning strategies that will allow aquaculture researchers andprofessionals to develop and implement improved culture practices.

Larval Fish Nutrition is divided

into three sections. The first section looks at the role of specific nutrient requirements in the healthy
digestive development of fish. The second section looks at the impacts if nutritional physiology on fish
through several early-life stages. The final section looks at feeding behaviors and the benefits anddrawbacks to both live feed and microparticulate diets in developing fish.

Langdon, C.L., 2003. Microparticle types for delivery of nutrients to marine fish larvae. Aquaculture
227, 259-275.
Good review and discussion of several types of microparticulate diets including complex
microparticles.

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Ronnestad, I., M. Yufera, B. Ueberscchar, L. Ribeiro, O. Saele and C. Boglione. 2013.

Feedingbehaviour and digestive physiology in larval fish: Current knowledge and gaps and bottlenecks inresearch.

Reviews in Aquaculture (suppl. 1) S59-S98.

Food uptake follows rules defined by feeding behaviour that determines the kind and quantity offood ingested by fish larvae as well as how live prey and food particles are detected, captured andingested. Feeding success depends on the progressive development of anatomical characteristics andphysiological functions and on the availability of suitable food items throughout larval development. The
fish larval stages present eco-morpho-physiological features very different from adults and differ from one
species to another. The organoleptic properties, dimensions, detectability, movements characteristics andbuoyancy of food items are all crucial features that should be considered, but is often ignored, in feeding
regimes. Ontogenetic changes in digestive function lead to limitations in the ability
to process certain feedstuffs. There is still a lack of knowledge about the digestion and absorption ofvarious nutrients and about the ontogeny of basic physiological mechanisms in fish larvae, including how
they are affected by genetic, dietary and environmental factors. The neural and hormonal regulation of thedigestive process and of appetite is critical for optimizing digestion. These processes are still poorly
described in fish larvae and attempts to develop optimal feeding regimes are often still on a ‘trial and error’basis. A holistic understanding of feeding ecology and digestive functions is important for designing dietsfor fish larvae and the adaptation of rearing conditions to meet requirements for the best presentation of
prey and microdiets, and their optimal ingestion, digestion and absorption. More research that targets gaps
in our knowledge should advance larval rearing.

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UJNR Annotated Bibliography

GENETICALLY MODIFIED SALMON IN AQUACULTURE; WELL REGULATEDAND SAFE
Olin, P
Van Eenennaam, Alison L. and Muir, William M. 2011. Transgenic salmon: a final leap to thegrocery shelf? Nature Biotechnology, Volume 29 number 8, Pp. 706-710

This paper presents a detailed analysis of the regulatory and review process that FDA
used for the AquAdvantage® salmon.

Entis, Elliot and Greenberg, Paul. 2013. The Salmon Dialogue: What’s the future of sustainable,
genetically engineered foods? Genetic Literacy Project.
http://www.geneticliteracyproject.org/2013/09/17/the-salmon-dialogue-whats-the-future-of-sustainab
le-genetically-engineered-foods/#.UjpTgtLBOSo

This online debate features, Elliot Entis, whose company has created a geneticallymodified salmon that may soon be for sale in the U.S., who discusses the environmental andhealth impacts of this controversial technology with author Paul Greenberg, a critic of GM
fish.
Menozzi, Davide, Mora, Cristina and Merigo, Alberto. 2012. Journal of Agrobiotechnology
Management & Economics, Volume 15, Number 3.

In this paper the authors provide an overview of salmon farming and world markets andthen analyze three scenarios based on the level of acceptance of GM salmon in themarketplace. The three scenarios were named as 1) ‘no market for GM fish,’ 2) ‘GM salmon
for dinner,’ and 3) ‘GM salmon doesn’t take off.’ The authors provide a summary of the main
outcomes for each scenario.
FDA 2012. AquAdvantage® Salmon, Draft Environmental Assessment. Center for Veterinary
Medicine, United States Food and Drug Administration, Department of Health and Human Services.
Washington D.C.

This document provides a detailed environmental assessment of the transgenic
AquAdvantage® Salmon

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LONG-TERM OUTCOMES IN THE TECH TRANSFER OF SCALLOP SPATCOLLECTION TECHNIQUES, FROM AOMORI PREFECTURE, JAPAN TO
MAINE, USA
Morse, D
Beal, B.F., M. Hastings, D. Morse, L. Taylor, T. Pottle, S. Rappaport, K. Porter, S. Inches, W.
Hopkins, J. Trenholm.

1999.

The Culture of Japanese Scallops (Patinopecten yessoensis), Reportof the Maine Delegation of Aomori Prefecture, Japan, May 14-21, 1999.

Maine Aquaculture

Innovation Center, Orono, Maine, USA.
The state of Maine (USA) shares a sister-state relationship with Aomori Prefecture in Japan, and thetwo have shared many exchanges over the years. In 1999, a group of Maine fishermen, scientists,
resource managers, processors and outreach staff travelled to Aomori to learn about the Japanese scallop
industry. Visits were made to scientific and research facilities, processing plants, government offices,
fishermen’s cooperatives and scallop farms.

Particular attention was paid to scallop spat collection

techniques and the supporting science, the seamless relationship between fishing and farming for seafood
producers in Mutsu Bay, and the various products made from scallops. Delegates from Maine found thetechniques for spat collection to be potentially very useful, and were impressed by the unity of purpose inhigh-quality scallop production from all sectors of the Japanese industry and government. Unity of
purpose is likely to be an issue in Maine, although the successful model of production, integration offishing and farming, and variety of products is a useful one as Maine considers the future of its’ scallop
fishery.

National Public Radio story on recent efforts in scallop aquaculture:
http://www.npr.org/2012/08/12/157674235/maine-lobstermen-give-farming-sea-scallops-a-try
In 2011, a pilot project in aquaculture of the Giant Sea Scallop (Placopecten magellanicus) was
initiated in nearshore Maine waters. The effort is unusual, in that growout sites and culture activities areoperated by both commercial fishermen and shellfish aquaculturists.

This overlap of industries isrelatively novel, but is paying rewards thus far. Initial observations of scallop growth are extremely
positive, and this radio piece by National Public Radio highlights both the production issues being
addressed as well as the opportunities that arise when fishermen and shellfish growers collaborate.

Web pages in support of scallop aquaculture trials, with attendant resources:
http://www.seagrant.umaine.edu/research/projects/dv/scallop-trials
The Maine Sea Grant program hosts many Web pages on various topics, as part of their activities in37

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extension and education. This page is one of several operated by Dana Morse of Maine Sea Grant, aspart of a series entitled ‘Resources for Shellfish Growers.’ Pages address a variety of topics, including
regulations and permitting, marketing, production aspects of several species, and production technology
generally. The page referenced above is tailored to an ongoing aquaculture production trial for the Giant
Sea Scallop (Placopecten magellanicus), as well as a source of several informative documents and othermedia, related to scallop aquaculture.

Dana’s Travelling Shellfish Blog
http://danastravelingshellfishblog.blogspot.com/
This blog is a means to transfer information to interested readers about shellfish and seaweedfarming, and the people in the aquaculture industry. The writing style is informal, but with enoughtechnical detail to be interesting and useful to those with some knowledge of the industry. Photos andvideos are included, and the blog also serves as a mechanism to report on the projects undertaken on the
job at Maine Sea Grant, including projects involving the production of sea scallops (Placopecten
magellanicus), sugar kelp (Saccharina latissima) and other seaweed species, razor clams (Ensis directus)
and blue mussels (Mytilus edulis).

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