United States-Japan
Natural Resources Panel on Aquaculture
40th Scientific Symposium“Hatchery Technology for High Quality Juvenile Production”
October 22-23, 2012
Honolulu, Hawai’i
Annotated Bibliography

ADVANCES IN DIAGNOSIS AND MANAGEMENT OF AMYLOODINIOSIS IN
INTENSIVE FISH CULTURE
Blaylock, RAnderson, R. M. and May, R. M. 1979. Population biology of infectious diseases: Part I.
Nature 280:361-367.
This is the foundational paper that established the modern concept of epidemiologicalmodelling. The paper argues for viewing the host population as a dynamic variable so as to unite
the medical and ecological view of diseases. As such, the host is viewed as a habitat for theparasite, the degree to which the parasite diminishes the reproductive capability of the host is
considered, the importance of acquired immunity is considered, and the degree to which host life
cycle structure determines transmission from one host to the next is incorporated. The result is aframework for building a simple model that can be tested with real world data.
Lotz, J. M. and Soto, M. A. 2002. Model of white spot syndrome virus (WSSV) epidemics in
Litopenaeus vannamei. Dis. Aquat. Org. 50:199-209.
The authors present a methodology for experimentally illuminating the population
dynamics of a significant pathogen of farmed shrimp. The paper first develops a generalized
diagrammatic view of the various states of the host population and the manner in which the virus“flows” through or connects the states of the host population. Connections are then transformed
into a series of mathematical equations and a laboratory challenge system is used to provide
experimental estimates of the rates of those connections. The authors evaluate the relative
importance of the various connections in the progress of an epidemic and conclude that athreshold density of susceptible shrimp exists below which an epidemic will not occur. Increases
in transmission efficiency lower the threshold value while most other factors increase it.
Epidemic models suggest testable hypotheses that may contribute to the development of control
strategies for WSSV outbreaks.
Masson, I., Blaylock, R.B., and Lotz, J.M. 2011. Susceptibility and tolerance of spotted
seatrout, Cynoscion nebulosus, and Red Snapper, Lutjanus campechanus, to experimental
infections with Amyloodinium ocellatum. J. Parasitol. 97:577-585.
The authors adapt the Lotz and Soto approach to a stage-structured model of anectoparasite. Components are the life-history stages of the parasite and connections are thetransitions between stages. Two species’ ability to harbor the dinoflagellate are compared.
Experimental estimates of the transitions are achieved. There were no differences in infection
rates or lethal doses between the red snapper and spotted seatrout, but trophonts grew larger andremained attached longer to red snapper. The trophont lethal load was higher in spotted seatrout.

UJNR Annotated Bibliography
A proposed model reflected the preliminary experimental findings and provides a foundation forbuilding an epidemiological model.
Mori, Y. and Notomi, T., 2009. Loop-mediated isothermal amplification (LAMP): a rapid,
accurate, and cost-effective diagnostic method for infectious diseases. J. Infect. Chemother.
15:62-69.
The authors, one of whom published the original paper on LAMP, review the technology
and advantages of the technique compared to other techniques such as PCR and other DNA
amplification methods. The advantages include the possibility of using loop primers that increase
the sensitivity and reduce the reaction time of the assay, the increase in turbidity of positive
samples that allows the simple visual assessment of the results, the ability to add fluorescence toaugment simple endpoint detection, less sensitivity to inhibitory substances, and the lack ofnecessity for complicated laboratory equipment or highly skilled staff. The authors then review
how LAMP has been applied to the study of infectious diseases and argue that the techniqueoffers a significant opportunity for expanding access to simple, inexpensive, reliable diagnostic
tools in socially and economically challenged areas.
Paperna, I. 1984. Reproduction cycle and tolerance to temperature and salinity of
Amyloodinium ocellatum (Brown, 1931) (Dinoflagellida). Annal. Parasitol. Hum. Comp.
59:7-30.
This paper established the basic morphological and biological characteristics of theparasite and delineated the effects of salinity and temperature on parameters of the life cycle. It
was shown that the parasite could exist in a wide range of temperatures and salinities, but that theideal temperature and salinity 25C and 33 PSU (although salinity tolerance varies withtemperature). Deviations from the ideal conditions affected the development and reproductive
capacity of the parasite, but the parasite could survive until normal conditions were reestablishedand normal reproduction could resume. Thus, the paper demonstrated why Amyloodinium
ocellatum is should a troublesome pest.
Noga, E. J. 2012. Amyloodinium ocellatum. In: Fish Parasites. Pathobiology and Protection.
Eds: Woo, P.T.K. & Buchmann, K. CAB International (Cambridge, USA).
This paper gives an overview of the research conducted on A. ocellatum focusing on new
diagnostic methods developed (specific PCR primers targeting the 3’ end of the Large Subunit of
the rDNA gene complex), most common internal/external lesions associated with outbreaks (e.g.
anorexia, pruritis, gill hyperplasia, etc), protective/control measures, environmental treatments
and innate/acquired resistance. This paper concludes that a better understanding of theenvironmental conditions affecting parasite growth and survival is essential to improve control
strategies. Also, it highlights the importance of the development of highly specific and sensitiveUJNR Annotated Bibliography
tests to detect all the different isolates of the parasites particularly when there is low number ofparasites (or latent carriers).

PRE-SPAWNING CAROTENOID FORTIFIED DIETS IMPROVE REPRODUCTIVE
TRAITS OF CHANNEL CATFISH, ICTALURUS PUNCTATUS AND SUBSEQUENTPROGENY PERFORMANCE
Chatakondi, N
J. Sawanboonchun, W. J. Roy, D. A. Robertson and J. G. Bell. 2008. The impact of dietary
supplementation with astaxanthin on egg quality in Atlantic cod broodstock (Gadus
morhua, L.) Aquaculture 283:97-101
The authors conducted this study to address the sub-optimal levels of carotenoids
observed in farmed broodfish and theire effect on their reproductive performance. Hence, thisstudy was conducted to assess the effect of supplementing astaxanthin in broodfish diets toAtlantic cod broodfish for two months prior to spawning. The authors found that supplemental
diets increased the concentration of astaxanthin in eggs by 2-3 fold in the farmed brood fish.
Higher astaxanthin levels prompted an increase of 20% higher in number of eggs produced andincreased the fertilization by 47% compared to the control group. They also postulated that thisimprovement may be attributed to the efficacy of astaxanthin supplementation in stress reduction
to enhance the immune system. The benefits of astaxanthin supplementation to broodstock
prompted the authors to suggest that all hatcheries be advised to address the problem of low
astaxanthin concentration in the oocytes of pre-spawning broodfish. Implications for nutrient
fortification in pre-spawning diets in farmed brood fish be derived from information presented inthis paper.
T. Ytrestoyl and B. Bjerkeng, 2007. Dose response in update and deposition of
intraperitoneally administered astaxanthin in Atlantic salmon (Salmo salar L.) and Atlanticcod (Gadus morhua L.) Aquaculture 263:179-191.
The authors determined the dose-response of astaxanthin in plasma, muscle, liver, kidneyand skin after intraperitoneal administration of hgh doses of water dispersible astaxanthin in
Atlantic salmon and Atlantic cod. The present study shows that the route of administration has a
profound effect on the amount of astaxanthin that is taken up in the blood and deposited in themuscle. The authors suggested that their procedure would increase the concentration ofastaxanthin in broodfish by three times compared to the broodfish administered by dietarymeans. Further, the authors concluded that the limitations observed dietary means of astaxanthin
supplementation was not by the limited number of binding sites in the muscle as previously
thought rather the limited up-take by the gastrointestinal tract of the fish. The magnitude of the
response of Atlantic salmon was higher than that of Atlantic cod. The authors concluded thatUJNR Annotated Bibliographyhigh concentration of astaxanthin could be obtained in Atlantic salmon in a short time byintraperitoneal injection compared to supplemental feeding means to have a positive impact on
reproductive performance. Implications of carotenoid by intraperitoneal injection in broodfish
for rapid incorporation of this nutrient in fish in a short time was presented in this paper.
S. Kim, J. Song, K. Kim and K. Lee. 2012. Effects of dietary astaxanthin on innate
immunity and disease resistance against Edwardsiella tarda in Olive Flounder Paralichthys
olivaceus. The Israeli Journal of Aquaculture (Bamidgeh) 64:740-746.
The authors conducted this study with the aim of reducing or preventing the overuse ofantibiotics in aquaculture and approached by immunomodulatory effects of nutrients, especiallyastaxanthin as a dietary supplement. The authors also examined the effect of astaxanthin on
sparing vitamin C, non-specific immune response, disease resistance against pathogenic agentsand the immunostimulant efficiency in Olive Flounder. The authors conducted a 15-day feeding
trial with 0, 1, 2 and 3% Astaxanthin supplemented diet fed in replicated tanks with 30 fish pertank. At the end of the study, the fish from each treatment group were given an intra-peritoneal
injection with an Edwardsiella tarda suspension and mortalities were recoded daily for 8 days.
The authors documented the immune reponse of bacterial disease challenge in the form of nonspecific response: Nitro-blue-tetrazolium (NBT) assay, Liver superoxide dismutase (SOD), andvitamin C sparing due to the effect of astaxanthin fortification. The authors concluded that 3%
astaxanthin administration enhanced the non-specific immune response and reduced thecumulative mortality to E. tarda in Olive Flounder under controlled conditions. Implications
for nutrient derived immunity in fish is presented in this paper.
Scanini, V., H. Fernandez-Palacios, L. Robaina, T. Kalinowski and M. S. Izquierdo. 2010.
Reproductive performance of gilthead seabream (Sparus aurata L) fed two combined levelsof cartoenoids from paprika oleoresin and essential fatty acids. Aquaculture Nutrition 17(3):304-312
The authors fortified broodfish diets with natural sources of carotenoids to assess theireffect on reproductive performance of gilthead seabream. Two combined levels of paprika
oleoresin at 40 and 60 mg/Kg along with two levels of n-3 HUFA were assessed in triplicate
groups of broodfish for 6 weeks. The authors demonstrated that an increase in carotenod content
in eggs significantly improved the egg viability, hatching rates and fecundity. The authors
concluded that the increased fertility observed in this study was attributed to the antioxidant
function of carotenoids, which played a role in protecting sperm cells from lipid peroxidation,enhanced sperm motility and fertility of seabream eggs. Implications for paprika oleoresin
supplementation in broodfish diets enhance reproductive performance of seabream.

WITHERING SYNDROME: DISTRIBUTION, IMPACTS, CURRENT DIAGNOSTIC
METHODS AND NEW FINDINGSUJNR Annotated BibliographyFriedman, C
Research in my laboratory focuses on marine organismal health with a focus on infectious
diseases of marine invertebrates and the role of climate drivers (e.g. abalone withering syndrome
and oyster herpes virus). In addition to disease, we also evaluate other facilitators of abalone
population health and help develop restoration of impacted populations. Below I describe papers
that highlight this research with a focus on recent advances.
Withering Syndrome:
Wetchateng,T, Friedman, C.S., , Wight, N.A., Lee, P.-Y., Teng, P.H., Sriurairattana, S.,
Wongprasert, K., and Withyachumnarnkul, B. 2010. Withering syndrome in abalone
Halioits diversicolor supertexta. Diseases of Aquatic Organisms 90(1):69-76. (Note that both
Wetchateng and Friedman are First authors).
This manuscript documents the first observation of the withering syndrome bacterium inThailand, Taiwan and the Peoples Republic of China. The paper describes the pathogen and its
impact on the host. Importation records combined with samples analyzed suggested that thebacterium was imported into Thailand via live animal transfers from California, USA initially to
China and Taiwan. The paper discusses the need for health examinations prior to animal
movements and the use of diagnostic methods to reduce the chance of introducing pathogens
with animal movements.
Braid, B. A., J. D. Moore, T. T. Robbins, R. P. Hedrick, R. S. Tjeerdema, and C. S.
Friedman. 2005. Health and survival of red abalone, Haliotis rufescens, under varying
temperature, food supply, and exposure to the agent of withering syndrome. Journal of
Invertebrate Pathology 89(3):219-231.
Braid et al. (2005) represents the M.S. thesis of Beverly Braid, a graduate student of minefrom UC Davis (originally co-advised by Dr. R. Hedrick and me but when I moved to UW, I
became a committee member). This paper was one of a two part collaborative project examining
withering syndrome between the Tjeerdema, Hedrick and Friedman laboratories:
A key element in understanding the disease physiology involved our discovery thatrickettsia-infected abalone require a thermal stress to develop clinical withering syndrome.
Although this fact was previously demonstrated, in Moore et al. (2000) we used abalone that had
been infected in situ and did not examine transmission as well. In Braid et al. (2005), we exposed
red abalone to the same does of the withering syndrome rickettsia and examined differential
transmission, pathogenesis, and survival at two environmentally relevant temperatures. Only 1.7%of the abalone were infected when held at ambient temperature (12.3oC), while 72-94% of those
held at 18.7oC became infected. In addition, we discovered that food availability significantly
influenced the proportion of exposed abalone that became infected with the withering syndrome
rickettsia; infection intensity was similar between these two treatments. Surprisingly, abalone fed
kelp developed more parasite-induced metaplasia than did starved individuals. However, losses
were higher in infected, starved abalones than those that were fed. Starved abalone at bothUJNR Annotated Bibliography
temperatures experienced high mortality than did fed animals and when held at the elevatedtemperature, the withering syndrome bacterium exacerbated losses induced by starvation andelevated water temperatures (e.g. as during an ENSO event).
This paper also investigated physiological aspects of disease development including
mechanisms of cellular change and energy reserves. We determined that depletion of glycogenreserves occurred in the digestive gland and foot muscle prior to pedal catabolism and visible
atrophy. In addition, we used two methods to try and identify the mechanism of metaplasticchanges observed in the digestive gland and revealed that neither elevated cell proliferation nor
apoptosis were involved.
These studies were complemented by a related manuscript using a metabolomic approachto assess biochemical changes in infected and uninfected red abalone (Robenblum et al. (2005).
An extension of our investigation into the role of climate change on abalone health is found in:Vilchis, L.I., Tegner, M.J., Moore, J.D., Friedman, C.S., Riser, K.L., Robbins, T.T., andDayton, P.K.. 2005. Effects of ocean warming on the growth, reproduction and survivorshipof red and green abalones in southern California. Ecological Applications 15(2):469-480.
Red and green abalone were exposed to varying temperature conditions (ambient + 3oC) ina withering syndrome endemic zone. When red and green abalone were exposed to thermal stress(ocean warming), red abalone reproduction was compromised green abalone were unaffected. In
addition, red abalone are more susceptible to withering syndrome than are green abalone (Vilchis
et al. 2005). This collaborative research has resulted in a more thorough understanding of the
impacts of withering syndrome on abalone and the effects of climatic variation of the potential
impact of this disease on abalone populations.
Another significant contribution to our understanding of this disease deals with its control
in commercial aquaculture or captive rearing settings through the used of an antibiotic via a seriesof manuscripts (Friedman et al. 2003, 2007, Rosenblum et al. 2005).
Friedman, C.S., Trevelyan, G., Mulder, E.P., and Fields, R. 2003. Development of an oral
administration of oxytetracycline to control losses due to withering syndrome in cultured
red abalone Haliotis rufescens. Aquaculture 224(1-4):1-23.
This research provided data for the compassionate use of the oxytetracycline to clear
rickettsial infections in abalone via the US Food and Drug Administration – Center for Veterinary
Medicine Program for Investigational New Animal Drugs. We demonstrated that the therapeutant
was rapidly depleted from the pedal muscle of medicated abalone and the participating farm wasgiven a 30 day withdrawal period. However, we later investigated differential clearance ofoxytetracycline from other tissues and demonstrated that the antibiotic was retained in thedigestive gland for prolonged periods (months!) in:UJNR Annotated Bibliography
Rosenblum, E.S., Robbins, T.T., Scott, B.B, Nelson, S., Juhasz, C., Craigmill, A.,
Tjeerdema, R.S, Moore, J.D, and Friedman, C.S. 2008. Efficacy, tissue distribution, andresidue depletion of oxytetracycline in WS-RLP infected California red abalone Haliotis
rufescens. Aquaculture 277:138-148.
In these experiments, abalone were medicated using 100mg/kg oxytetracycline for varying
timer periods to examine not only treatment efficacy and depletion but also protection from reinfection. Levels of oxytetracycline accumulated and retained in abalone digestive gland was far
greater than ever reported in the literature in any other animal investigated (marine or terrestrial).
Based on this data, the farm mentioned above stopped all treatments. We also examined the levelof divalent cations in abalone digestive gland and hypothesized that the observed high metal
concentrations may be responsible for the long retention time of this drug in our animals. Using
our re-challenge studies we also demonstrated that low levels of oxytetracycline are protective
against exposure to the withering syndrome rickettsia, further optimizing the use of thistherapeutant in a judicious manner. Although the long depletion time for this drug in abalone isproblematic from a commercial culture and environmental stand point, it provides an opportunity
for captive rearing of endangered abalone. A single treatment series may protect the animals formonths if reared in waters in which the bacterium is present; the withering syndrome rickettsia
ranges from Mexico north to San Francisco (Friedman and Finley 2003) and has been found in two
locales in northern California where (infected) abalone were out-planted prior to the identificationof the causative agent of this disease. In fact, our treatment has been used to protect endangered
white abalone broodstock, to reduce losses in captive-bred juvenile white abalones and is being
incorporated into the NMFS white abalone management plan as required by the Endangered
Species Act (Friedman et al. 2007).
This information not only aids in the control of this disease but has allowed us to better
examine the host physiology and pharmacokenetics of this drug in abalone. In fact, abalones
appear to have the longest retention of oxytetracycline of any animal examined to date, which is
crucial information for the aquaculture industry and US-FDA/CVM, the agency that regulates drug
approval in the USA.
Ostreid (Oyster) Herpesvirus
Another important area of research in my laboratory deals with health of the Pacificoyster, an important species within our US west coast aquaculture industry. We have developed
a collaborative program with researchers in the USA and France to better investigate causes ofoyster mortality with a focus on early life stages and the influence of pathogens and climate on
disease expression.
Burge, C.A., Griffin, F.J., and Friedman, C.S. 2006. Summer mortality and herpes virus
infections of the Pacific oyster, Crassostrea gigas, in Tomales Bay, California. Diseases ofAquatic Organisms 72:31-43.

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This collaborative research complements Burge et al. (2007) in which our research team aswell as Dr. Renault (IFREMER, France) assessed the influence of site, planting time, oyster line
and climatic conditions (temperature and salinity), and phytoplankton assemblages on Pacificoyster survival. The present appear used a combination of field and laboratory studies to evaluate
the influence of oyster stock, seed size, site, and oyster herpesvirus (OsHV) presence on survivaland growth of Pacific oyster seed. We demonstrated that OsHV was present in moribund oysters
and that it’s presence predicted seed oyster mortality. In addition, differential losses wereobserved among oyster stocks planted in the bay. One family produced from wild stock from
Washington state, routinely out-performed other commercial stocks or selected family lines
employed in this study, thereby providing oyster culturists will important information forsustainability of their industry. Seed size was inversely correlated with survival, while temperature
and OsHV presence directly correlated with losses suggesting a synergistic effect of these twostressors on disease expression. We demonstrated that cellular changes suggestive of herpesviruses
were rare and suggest that multiple diagnostic methods are needed to examine the influence of
OsHV on seed oyster survival.
This paper provided the foundation of our more recent OsHV research designed to better
understand the ecology of this disease: we developed a quantitative, real-time PCR assay to detect
OsHV in oyster and water samples. Using this assay as well as histological analyses wedemonstrated that although viral DNA was detected in several bivalves in Tomales Bay (C. gigas,
Ostrea edulis, C. virginica, C. sikamea, Mytilus galloprovincialis, and Venerupis phillipinarum),significantly higher OsHV copy numbers were observed in the Pacific oyster (Burge et al. in
review). This research is complemented by several other papers by Burge et al. 2007, 2010, 2011.

FEEDING HATCHERY-PRODUCED LARVAE FROM THE GIANT GROUPEREPINEPHELUS LANCEOLATUS
García-Ortega, A
Tucker, J.W. and Woodward, P.N.. 1996. Nassau grouper aquaculture. Pp. 363-377. In: F.
Arreguin-Sanchez, F., Munro, J.L., Balgos, M.C. and Pauly, D. (eds.) Biology, fisheries andculture of tropical groupers and snappers. ICLARM Conf. Proc. 48. 449 p.
The authors of this paper review multiple spawning, larval rearing and growout trials for
the Nassau grouper (Epinephelus striatus). They were able to successfully spawn the grouper bymultiple methods including combinations of induced (HCG) and natural ovulation along withartificial and natural fertilization. During multiple larval rearing trials using different size tanks,
feeding regimes, and other parameters the best survival recorded was 5% from fertilization to 98
dph. During growout grouper reached a mean weight of 1.5 kg at 23 months with a respectiveUJNR Annotated Bibliography
food conversion ratio of 0.9-1.3. At 28 months the mean weight was 2.0 kg with a foodconversion ratio of up to 1.8.
Colin, P.L., Koenig, C.C. and Laroche, W.A. 1996. Development from egg to juvenile of the
red grouper (Epinephelus morio) (Pisces: Serranidae) in the laboratory. Pp. 399-414. In: F.
Arreguin-Sanchez, F., Munro, J.L., Balgos, M.C. and Pauly, D. (eds.) Biology, fisheries andculture of tropical groupers and snappers. ICLARM Conf. Proc. 48, 449 p.
This paper represents one of the first successful spawning and larval rearing trials for the
red grouper (Epinephelus morio). This paper offers an in depth description of larval developmentof this grouper from egg to juvenile focusing on morphometrics, pigmentation and behavior.
Furthermore, this paper describes how salinity and temperature affects growth and development.
Eggs averaged 0.95mm diameter and hatched in ~30 hours at 24°C with metamorphosis occurred
starting 35 days post hatch at a size of 20 mm standard length.
Kiriyakit, A., Gallardo, W.G. and Bart, A.N. 2011. Successful hybridization of groupers(Epinephelus coioides x Epinephelus lanceolatus) using cryopreserved sperm. Aquaculture
320:106-112.
In this recent paper the larval rearing of a grouper hybrid from E. coioides eggs and E.
lanceolatus sperm is described. Feeding of hybrid larvae was progressively done using oyster
trocophore larvae, rotifers and Artemia. This is an important paper because little if any
information exists in the larval rearing and development of the giant grouper. In addition, thispaper reports rotifers were used without copepods in the successful larval rearing of this hybrid,
which was not achieved in our study with E. lanceolatus. Additional information is provided on
the larval development and metamorphosis of the hybrid fish that in some aspects is similar tothe developmental stages we observed during in the larval rearing of E. lanceolatus.

STUDY ON HIGH INCIDENCE OF DEATH DUE TO COLLISION OF HATCHERYREARED PACIFIC BLUEFIN TUNA THUNNUS ORIENTALIS JUVENILES IN NET
CAGES
Higuchi, K
Miyashita, S., Sawada, Y., Hattori, N., Nakatsukasa, H., Okada, T., Murata, O. and Kumai,
H. 2000. Mortality of northern bluefin tuna Thunnus thynnus due to trauma caused bycollision during growout culture. Journal of the World Aquaculture Society, Vol 31. Pp
632-639.

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Collisions with the walls of tanks or nets caused high mortality of northern bluefin tuna
during juvenile stage. In this study, juveniles were reared in indoor tanks from 30 to 120 day
post-hatching (dph) and in a sea net cage from 42 to 150 dph, and dead fish were examined todetect injury of the bones using x-ray. As a result, more than 98% fish were dead until end ofboth experiments. The proportion of dead fish with injuries of bone, especially of the vertebral
column and the parasphenoid, increased after fish reached 50 mm in BL (35 dph), and exceeded
60% in fish with BL 85 mm (40 dph) or greater in the indoor tanks. Also, the injuries of thevertebral column and the parasphenoid were observed in most of dead fish in the net cage. These
results show that the loss of juvenile bluefin tuna was caused by collision with the tank or net
wall that fatally damaged the bones of the vertebral column and the parasphenoid.
Masuma, S., Kawamura, G., Tezuka, N., Koiso, M. and Namba, K. 2001. Retinomotor
responses of juvenile bluefin tuna Thunnus thynnus. Fisheries Science, Vol 67. Pp 228-231.
In bluefin tuna culture, a high mortality of juveniles is caused by collision with the tankand net cage walls at dawn. This collision can possibly be attributed to visually disoriented
behavior of the fish. To examine this possibility, the authors carried out retinomotor response
experiments with juvenile bluefin tuna and measured ambient light intensity at the culture site at
dawn. The light intensity at which the transition from scotopic to photopic vision takes place was
7.52 lx and the time taken by the transition was 15 min. At dawn, the ambient light intensity
rapidly increased from scotopic light intensity level and attained photopic light intensity level in
10 min. This incompatibility of the retinal adaptation with the change in the ambient lightintensity could cause the visual disorientation of the fish. It is therefore possible that the visuallydisoriented juveniles cannot control their high power swimming and thus collide with the walls
at dawn.
Ishibashi, Y., Honryo, T., Saida, K., Hagiwara, A., Miyashita, S., Sawada, Y., Okada, T.
and Kurata, M. 2009. Artificial lighting prevents high night-time mortality of juvenile
Pacific bluefin tuna, Thunnus orientalis, caused by poor scotopic vision. Aquaculture, Vol
293. Pp 157-163.
In a lab-based study using infrared camera, death of juvenile Pacific bluefin tuna (PBT)
was associated with touching and/or colliding with the net wall attached to an indoor tank, thenexhibiting abnormal swimming behavior, then becoming moribund during the night and dawn.
Also, the scotopic visual threshold of juvenile PBT was at least 40 fold inferior to that of juvenile
red sea bream, ocellate puffer, purplish amberjack and grouper. To make up to low scotopic
vision sensitivity of juvenile PBT and prevent juvenile PBT from touching and/or colliding with
the net wall, the authors illuminated sea net cages by overhead lights providing 200-3000 lx
intensity at the water surface. As a result, survival rate of juvenile PBT in the sea net cages
significantly improved to 73% up to 23 days post-transfer compared to 12% survival in non-lightcontrol net cage.

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SUCCESS OF SEED PRODUCTION OF HUMPHEAD WRASSE CHEILINUS
UNDULATUS WITH IMPROVEMENT OF SPAWNING INDUCTION, FEEDING, AND
REARING CONDITION
Hirai, N
Hirai, N., M. Koiso., K. Teruya., M. Kobayashi., T. Takebe., T. Sato., K. Nakamura., T.
Goto., and A. Hagiwara. 2012. Rearing conditions for Humphead Wrasse Cheilinus
undulatus Larvae, and Introduction of the Minute Rotifer Proales similis as an Initial Live
Food. Journal of Fisheries Technology, Vol 4:2, Pp 57-64. (in Japanese with English abstract)
Fisheries Research Agency and Nagasaki University. 2011. Japan’s first production of
humphead wrasse fry – World’s first reproducible production – (in Japanese), Press
Release, URL: http://www.fra.affrc.go.jp/pressrelease/pr23/231129/index.html
Wullur S, Sakakura Y, and Hagiwara A (2009) The minute monogonont rotifer Proales
similis de Beauchamp: Culture and feeding to small mouth fish larvae. Aquaculture, 293,
62-67.
WITHERING SYNDROME IN ABALONE IN JAPANKiryu, I
Kiryu, I. 2011. Aqua Culture Magazine, Midori Shobo Co. Pp 86.

EFFECTS OF DOCOSAHEXAENOIC ACID AND TAURINE LEVELS IN ROTIFERS
ON GROWTH AND SURVIVAL OF LARVAL AMBERJACK SERIOLA DUMERILI
Matsunari, H
Yamamoto T, Teruya K, Hara T, Hokazono H, Hashimoto H, Suzuki N, Iwashita Y,
Matsunari H, Furuita H, Mushiake K 2008. Nutritional evaluation of live food organisms
and commercial dry feeds used for the seed production of amberjack Seriola dumerili. FishSci 74,1096-1108
To improve the nutritional quality of live foods and dry feeds ordinarily used for the seedproduction of amberjack Seriola dumerili, the nutrient contents of rotifers, Artemia nauplii andcommercial feeds used in two larval production stations were evaluated. For comparison of thenutrient contents, artificially produced larvae, wild-caught juveniles and wild zooplankton
samples were also analyzed. The proportions of 22:6n-3 in the polar lipid of the cultured larvae
increased by feeding the dry feeds. The taurine contents of the cultured larvae reflected the

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contents of their foods(rotifers < dry feed < Artemia nauplii). The taurine content and the
proportion of 22:6n-3 in Acartia spp. were higher than in foods fed to the larvae. These
parameters in the wild juveniles were higher than the cultured ones.
Matsunari H, Hashimoto H, Oda K, Masuda Y, Imaizumi H, Teruya K, Furuita H,
Yamamoto T, Hamada K, Mushiake K 2012. Effects of docosahexaenoic acid on growth,survival and swim bladder inflation of larval amberjack Seriola dumerili. Aquac Res
doi:10.111/j.1365-2109.2012.03174.x
The effect of DHA on the growth performance and survival of larval Seriola dumeriliduring the rotifer feeding period was investigated. Amberjack larvae at 3 day post hatching were
fed rotifers enriched with freshwater Chlorella (Chlo), a mixture (2:1, v/v) of Chlo and DHAenriched Chlorella (DHA-Chlo), DHA-Chlo, and DHA-Chlo and commercial DHA emulsion, in
triplicate for 7 days. The survival rate was improved by the enrichment of rotifers with DHAChlo alone, and DHA-Chlo and emulsion. Growth and swim bladder inflation of fish fed rotifers
enriched with DHA-Chlo were significantly improved, however, with increased levels of DHA
further improvement was not found. The DHA requirement of amberjack larvae is estimated tobe 1.5mg/g on a dry matter basis of rotifers.
Matsunari H, Hashimoto H, Oda K, Masuda Y, Imaizumi H, Teruya K, Furuita H,
Yamamoto T, Hamada K, Mushiake K 2012. Effect of different algae used for enrichmentof rotifers on growth, survival and swim bladder inflation of larval amberjack Seriola
dumerili. Aquac Int doi:10.1007/s10499-12-9522-8
The effect of algae with different DHA contents used for the enrichment of rotifers on thegrowth performance, survival, and swim bladder inflation of larval amberjack Seriola dumerili
was investigated. Rotifers were fed with freshwater Chlorella vulgaris containing 3 levels of
DHA (rotifer containing DHA 0.04, 0.60, 1.32g 100g-1) and Nannochloropsis (rotifer containing
DHA, 0.04g 100g-1; EPA, 2.54g 100g-1). The same algae were supplemented to the larval rearing
tanks in static condition. The larvae in each triplicate group were fed the enriched rotifers from 3days post-hatch for 7 days. Growth and survival rate of fish fed the rotifers enriched with
Nannochloropsis were higher than those of fish fed the rotifers enriched with all three Chlorella
treatments. Swim bladder inflation was lowest in fish fed the rotifers enriched with
Nannochloropsis.
Although rotifers enriched with Nannochloropsis were effective for the growth andsurvival, DHA is essential for swim bladder inflation in amberjack larvae.

APPLICATION TO THE GENETIC BREEDING USING GENOMICS INFORMATIONIN YELLOWTAIL (SERIOLA QUINQUERADIATA)UJNR Annotated Bibliography
Ozaki, A
Fuji K, Yoshida K, Hattori K, Ozaki A, Araki K, Okauchi M, Kubota S, Okamoto N,
Sakamoto T (2010). Identification of the sex-linked locus in yellowtail, Seriola
quinqueradiata. Aquaculture 308: 51-55
The sex-determining system of yellowtail (Japanese amberjack), Seriola quinqueradiata,
is not known. In thisstudy, we identified the sex-linked locus in yellowtail and we characterized
the sex-determining system by genetic linkage analysis conducted on 19 female and 19 maleprogenies from a single family. The associations between phenotypic sex and genotypic data of
71 microsatellite markers selected from yellowtail genetic linkage map were tested. The putative
sex-determining locus is located between locus Sequ21 and locus Sequ17 in LG12, and the sexlinked alleles were inherited from the female parent. This result suggests that yellowtail has a
ZZ–ZW sex-determining system, and that it would be possible to use these sex-linked markers todiscriminate the sexes.
Fuji K, Kai W, Kubota S, Yoshida K, Ozaki A, Aoki J, Kawabata Y, Araki K, Tsuzaki T,
Okamoto N, Sakamoto T (2012). A genetic linkage map of yellowtail (Seriola
quinqueradiata) constructed with microsatellite isolated from BAC-end sequences. BMC
Genomics: submitting
The Japanese amberjack/yellowtail (Seriola quinqueradiata) is a popular marine cultured
fish in Japan. Presently, the growing fish pens are stocked with wild-caught juvenile but using
cultured brood fish for seed would have less environmental impact. Therefore, it is requested tochange the seeds from wild to artificial ones, and allow the selection of commercially important
traits. Information on genetic markers and genomic large insert libraries associated with QTL canbe used to select individuals carrying desired traits in breeding programs and to identify
causative genes. In this study, we prepared a BAC library and present the female and malelinkage maps of yellowtail with microsatellite markers obtained from BAC-end sequences(BESs) and a yellowtail genomic library. The BAC library consists of 110,592 clones with anaverage insert size of 140.7 kb, representing a 16-fold coverage of the genome. From 5,324
BESs, 743 primer pairs were designed to amplify a mapping panel and 373 primer pairs wereidentified. A total of 464 microsatellite markers derived from a yellowtail genomic library werealso mapped. These 837 markers were mapped on female and male maps in 24 linkage groups.
The difference of recombination rates between the female and male maps was very small(Female:Male=0.99:1). All linkage groups in the maps indicated that the locations of the sexspecific recombination hot-spots were very different in males and females. Female and malelinkage maps of yellowtail were constructed using BAC-clones information. After a genomic
region has been shown to be linked to trait of economic importance, this high quality BAC
library resource and mapped BAC clones information are crucial in the identification andfunctional characterization of the genetic variation.

UJNR Annotated Bibliography
Ozaki A, Yoshida K, Fuji K, Kubota S, Kai W, Suzuki J, Akita K, Aoki J, Kawabata Y,
Nakagawa M, Hotta T, Tsuzaki T, Okamoto N, Araki K, Sakamoto T (2012). Quantitative
trait loci (QTL) associated with resistance to a parasitic disease (Benedenia seriolae) inyellowtail (Seriola quinqueradiata) through genome wide analysis. Heredity: submitting
Benedenia disease caused by the ectoparasite Benedenia seriolae is a serious parasitic
disease in marine aquaculture finfish, leading to secondary viral or bacterial infections. Because
fish rub their bodies against the fish cage to remove the parasite, the mortality is quite highespecially in juveniles. Also the method to remove requires a lot of time, cost and effort.
Benedenia disease is difficult to prevent in marine aquaculture systems. Genetic variation has
been indicated to play a significant role to determining the susceptibility to this parasitic disease,
however, the mechanisms involved in the differential response to infection remain poorly
understood. To evaluate the genetic basis of Benedenia disease resistance in yellowtail (Seriola
quinqueradiata), a genome-wide and chromosome-wide linkage analysis was initiated using F1yellowtail families. Two major quantitative trait loci (QTL) regions on linkage groups Squ2 andSqu20 were identified and then confirmed in F1 families. These QTL regions explained 32.9–
35.5% of the phenotypic variance. On the other hand, the QTL related to growth was found inother linkage groups (Squ7). Therefore, Benedenia disease resistance QTL was not correlatedwith fish size. At the results, we have discovered the first genetic evidence that contributes todetailing the phenotypic resistance to Benedenia disease, and the results will help resolve themechanism of resistance to this important disease of yellowtail.

THE COOPERATIVE CULTURE OF SEAWEED IN NEW ENGLAND—HOW
RESEARCH, INDUSTRY, AND EXTENSION ARE CULTIVATING A NEW FIELD IN
AQUACULTURE
Redmond, S
Benson, Judy. 2012, March 11. Seaweed farming in the Sound: The beginning of something
big? The Day. Retrieved fromhttp://www.theday.com/article/20120311/NWS01/303119895/1018
This Connecticut newspaper article summarizes the research work on native New
England seaweed species for farms in Maine and Long Island Sound. A collaborative research
project involving the University of Connecticut Marine Biotechnology Research Laboratory andcommercial partners Ocean Approved, Llc in Maine resulted in the first successful kelp culture
crops being cultivated both in Maine and Long Island Sound in the winter of 2011, with plans tofurther develop farming operations in the future.
Maine Sea Grant. 2012. Seaweed Production on Mussel Farms in Maine:UJNR Annotated Bibliography
A Pilot Project to Stimulate Seaweed Production on Mussel Farms in Maine [Press
Release] Retrieved from http://www.seagrant.umaine.edu/extension/kelp-mussels
This online press release from Maine Sea Grant summarizes the collaborative research
work conducted by shellfish growers, seaweed farmers, and marine extension agents in Maine.
The project was the first of its kind in Maine, integrating Saccharina latissima on six different
shellfish farms (mussel and oyster) in Maine.
Morrissey, MT & S Almonacid. 2005. Rethinking technology transfer. Journal of FoodEngineering, Volume 67, Issues 1–2, pp. 135-145.
(http://www.sciencedirect.com/science/article/pii/S0260877404003346)
This article uses several new seafood processing innovation examples to highlight theimportance of a new approach to technology transfer. In order to improve and compete in a
global marketplace, businesses need more real-time, adaptable approaches to technology andresearch being developed in labs and Universities. In order to keep the speed of transfer up withthe speed of new technology development, the authors suggest a more multi-disciplinary anddynamic approach.
Warner, KD. 2008. Agroecology as Participatory Science: Emerging Alternatives toTechnology Transfer Extension Practice. Science, Technology, & Human Values , Vol. 33,
No. 6, pp. 754-777.
Agricultural extension plays an important role of linking the research community with thebusiness community, transferring new technologies from scientists to farmers, and problems offarmers as research questions back to scientists. Besides just technology transfer, extension
plays a wider role as educators of the general public, integrating new technologies, producers,
consumers, and issues arising from these. This article reviews these interactions by looking atenvironmental pollution issues and agroecological partnerships between growers, extension, andscientists. This type of participatory science uses collaboration, multidisciplinary approaches,and ecological approaches to improve the direction of agricultural developments.
Parkhurst, Emily. 2011, September 26. Kelp wanted: Seaweed farms expand in Maine’s
Casco Bay. The Forecaster. Retrieved from http://www.theforecaster.net/content/pnmkelp-farm-near-chebeague-092811
This newspaper article reports on the expansion of the first commercial kelp farm in the
United States, Ocean Approved, Llc, in Maine. They are a new company with two years ofcultivation experience, with plans to expand their product lines and further develop seaweed
aquaculture in Maine. Part of their plan involves sale of their native “seed” plants to othergrowers in Maine.

UJNR Annotated Bibliography
INTENSIVE JUVENILE PRODUCTION OF YELLOWTAIL AMBERJACK (Seriolalalandi) IN SOUTHERN CALIFORNIA
Rotman, F
Attramadal, K.J.K., T. Bjørnar, I. Salvesen, G. Øie, O. Vadstein and Y. Olsen. 2012.
Ceramic clay reduces the load of organic matter and bacteria in marine fish larval culturetanks. Aquacultural Engineering Volume 49 Pp 23-34.
The authors of this paper describe how the use of ceramic clay as a replacement for live
microalgae or algae pastes can increase turbidity without contributing to the organic matter loadin larval marine fish rearing systems. A series of experiments is described that demonstrateencouraging results of the larval performance of Atlantic cod (Gadus morhua) reared with clay
as a turbidity source compared to those reared using standard live microalgae and paste
protocols.
Hernández-López, J., M.A. Guzmán-Murillo and F. Vargas-Albores. 1995. Quantificationof pathogenic marine Vibrio using membrane filter technique. J. of Microbiological
Methods Volume 21:2 Pp 143-149.
These authors describe a rapid and simple methodology based on a membrane filter
technique designed to detect and quantify marine Vibrio spp involved in fish and shellfish
diseases as well as human gastrointestinal disorders. This technique can be instrumental in the
management of bacterial communities in marine larval fish culture systems.
Olafsen, J.A. 2001. Interactions between fish larvae and bacteria in marine aquaculture.
Aquaculture, Volume 200 Pp 223-247.
This publication is an excellent review of the influence bacterial ecology can have on theculture of marine organisms. Specifically, the author reviews the interaction between various
bacterial species and larval marine fish. Understanding concepts discussed in this publication are
instrumental to the successful larval rearing of various marine organisms.
Yamamoto, T., K. Teruya, T. Hara, H. Hokazono, H. Hashimoto, N. Suzuki, Y. Iwashita,
H. Matsunari, H. Furuita and K Mushiake. 2008. Nutritional evaluation of live foodorganisms and commercial dry feeds used for seed production of amberjack Seriola
dumerili. Fisheries Science, Volume 74 Pp 1096-1108.
The authors of this influential paper compare the nutritional profiles of both wildplankton and wild Seriola dumerili juveniles to commercially produced dry diets, cultured live
feeds and cultured juvenile fish. Differences in resulting nutrient levels help to identify potential
deficiencies in cultured S. dumerili. More specifically, it was found that 22-6n-3 and taurine aremuch higher in wild copepods (Acartia spp.) and S. dumerili juveniles than the cultured
counterparts of both.

UJNR Annotated Bibliography

IN SITU SWIMMING AND SETTLEMENT BEHAVIOR OF CULTURED SERRANIDLARVAE, PLECTROPOMUS LEOPARDUS AND EPINEPHELUS MALABARICUS
Shibuno, T
Masuma, S., N. Tezuka and K. Teruya. 1993. Embryonic and morphological developmentof larval and juvenile coral trout, Plectropomus leopardus. Japanese Journal of Ichthyology,
40(3): 333-342.
At the Japan Sea Farming Association, Yaeyama Station, coral trout broodstock haveprovided fertilized eggs by spontaneous spawning since May 1988. They described theembryonic and morphologica1 development, behavior, and growth of the coral trout from theegg to the juvenile stages.
Leis, J. M. and B. M. Carson-Ewart. 1999. In situ swimming and settlement behavior oflarvae of an Indo-Pacific coral-reef fish, the coral trout Plectropomus leopardus (Pisces:
Serranidae). Marine Bio1ogy134: 51-64.
Late larvae of the serranid coral trout Plectropomus leopardus (Lacepède), captured inlight traps, were released during the day both in open water and adjacent to two reefs, and their
behaviour was observed by divers at Lizard 1sland, northern Great Barrier Reef.
Nakai, T. 2002. Management of fishery resources for groupers (Serranidae) in Okinawa,
southern Japan. Fisheries Science, 68(Supplement): 431-432.
They show data on the distribution of groupers across the coral reef area at Iriomote
Island, and a summary on the seed production of several species of grouper in the Ryukyu Island,
Okinawa. Their data on the distribution of groupers provide additional information on the
management strategy for groupers.
Shibuno, T., Y. Nakamura, M. Horinouchi and M.Sano. 2008. Comparison of reef fish
community structures from mangrove estuary to coral reef slope, at Ishigaki Island,
southern Japan. Ichthyological Research. 55(3):218-237.
To clarify seascape-scale habitat use patterns of fishes in the Ryukyu Islauds (southern
Japan), visual censuses were conducted in the mangrove estuary, sand area, seagrass bed, coral
rubble area, branching coral area on the reef flat, and tabular coral area on the outer reef slope.

EFFECTS ON GROWTH OF FLATFISH JUVENILE BY ARTIFICIAL LIGHTENING
CONDITIONS: INTENSITY, PHOTOPERIOD AND WAVELENGTHUJNR Annotated Bibliography
Shimizu, D
Itoh, K., Y. Washio, Y. Fujinami, D. Shimizu, S. Uji, H. Yokoi, T. Suzuki. 2012. Continuous
illumination through larval development suppresses dopamine synthesis in the
suprachiasmatic nucleus, causing activation of α-MSH synthesis in the pituitary andabnormal metamorphic skin pigmentation in flounder. General and Comparative
Endocrinology 176: 215–221.
In order to better understand the endocrine aberrations related to abnormal metamorphic
pigmentation that appear in flounder larvae reared in tanks, this study examined the effects ofcontinuous 24-h illumination (LL) through larval development on the expression of tyrosinehydroxylase-1 (th1), proopiomelanocortin (pomc), α-melanophore-stimulating hormone (αMSH) and melanin concentrating hormone (MCH), which are known to participate in the controlof background adaptation of body color. We observed two conspicuous deviations in theendocrine system under LL when compared with natural light conditions (LD). First, LL
severely suppressed th1 expression in the dopaminergic neurons in the anterior diencephalon,
including the suprachiasmatic nucleus (SCN). Second, pomc and α-MSH expression in the parsintermedia melanotrophs was enhanced by LL. Skin color was paler under LL than LD before
metamorphic pigmentation, and abnormal metamorphic pigmentation occurred at a higher ratioin LL. We therefore hypothesize that continuous LL inhibited dopamine synthesis in the SCN,
which resulted in up-regulation of pomc mRNA expression in the melanotrophs. In spite of the
up-regulation of pomc in the melanotrophs, larvalskin was adjusted to be pale by MCH which
was not affected by LL. Accumulation of α-MSH in the melanotrophs is caused by uncoupling of
α-MSH synthesis and secretion due to inhibitory role of MCH on α-MSH secretion, which
results in abnormal metamorphic pigmentation by affecting differentiation of adult-typemelanophores. Our data demonstrate that continuous illumination at the post-embryonic stage
has negative effects on the neuroendocrine system and pituitary in flounder.

LARVAL REARING ADVANCEMENTS FOR YELLOWTAIL AMBERJACK (Seriolalalandi) IN SOUTHERN CALIFORNIA
Stuart, K
Attramadal K.J.K., B. Tondel, I. Salvensen, G. Oie, O. Vadstein, Y. Olsen. 2012. Ceramicclay reduces the load of organic matter and bacteria in marine fish larval culture tanks.
Aquaculture Engineering 49: 23-34.
The authors present information on the use of ceramic clay as a turbidity source formarine larvae. In contrast to live microalgae or algae pastes, clay increases turbidity without
contributing to the organic matter load. The larvae, algae, and live feed increase the microbial
carrying capacity of the rearing water which allow exponential growth of bacteria and favor fast-UJNR Annotated Bibliography
growing opportunists. Reducing substrate levels by replacing microalgae with clay may reducebacteria proliferation and benefit larvae. The authors compared the effects of three rearing
regimes including live Isochrysis galbana, Nannochloropsis oculata paste, and ceramic clay on
the bacterial community, concentration of organic matter, and growth and survival of Atlantic
cod larvae (Gadus morhua L.). The application of clay resulted in reduced substrate levels forbacteria in the rearing water compared to the addition of live algae or algae paste. The tanksreceiving clay showed a lower bacterial level in the water than tanks that added algae paste orlive algae. The algae paste treatments showed a higher abundance of bacteria and a higher shareof cultivable bacteria and TCBS counts than the other two treatments. Tanks with live algae
showed low relative abundances of opportunistic bacteria and TCBS counts in both water androtifers. Larval growth was significantly higher in the clay and live algae treatments than thealgae paste treatment. The survival of larvae in the tanks added clay was variable. Two of thethree tanks with clay had significantly higher larval survival than the tanks with live algae oralgae paste. However, one tank with clay underwent 100% mortality. The authors could not
explain this variation but mention that the effects of clay addition on larval performance should
be studied further. Clay addition appears to be an easy way to reduce bacterial load during early
first feeding of marine larvae without compromising the beneficial effects of turbidity.
Cook, M.A., R.B. Johnson, P. Nicklason, H. Barnett, M.B. Rust. 2008. Marking live feeds
withinert metal oxides for fish larvae feeding and nutrition studies. Aquaculture Research 39:
347-353.
The authors used yttrium oxide (Y2O3), ytterbium oxide (Yb2O3), lanthanum oxide(La2O3) and dysprosium oxide (By2O3) as potential live feed markers for feeding and nutrition
trials with fish larvae. These markers were evaluated by determining the uptake and depletion ofmarkers over time and quantifying ingestion of yttrium oxide marked rotifers by Atlantic cod(Gadus morhua). The authors found that Artemia nauplii and rotifers quickly took up markers
within 10 minutes and that there was no significant difference among temperatures in depletionof markers (10, 15, and 20 C) with Artemia or rotifers. Depletion from rotifers was not
significantly different between 5 and 20 minutes or between 5 and 30 minutes for Artemia. The
authors were able to show that visual Artemia estimates consumed by the larvae were similar to
consumption estimates determined by analysis of yttrium oxide marked Artemia using
inductively coupled plasma optical emission spectroscopy.
Stuart, K.R. and M.A. Drawbridge. 2012. Captive spawning and larval rearing of
California yellowtail (Seriola lalandi). Aquaculture Research doi:10.1111/j.13652109.2011.03077.
The authors document spawning patterns, measures of egg production, population
fecundity, and egg and larval quality over a four year period for a captive population of Seriolalalandi. Spawned eggs were also used to document larval development and to develop rearingUJNR Annotated Bibliography
techniques for aquaculture in the region. Authors showed that spawning occurred naturally inthe 140 m3 tank when the ambient water temperature reached 16°C and ended when thetemperature exceeded 22°C and egg production reached a maximum when 43 spawn events wererecorded from a pool of nine females yielding 36.8 million eggs total. Larval rearing trials
yielded survival rates as high as 5.8% from egg to 50 dph. Successful larval culture methods
included the addition of algae paste for green water culture, rotifers (20 rotifers ml-1) at 2 dph
and Artemia (5 Artemia ml-1) at 6 dph. Larvae were transferred from the incubation tank at 10
dph to a shallower tank with 33% greater surface area in order to accommodate the larvae’s
strong orientation to surface waters.
Benetti, D. 1997. Spawning and larval husbandry of flounder (Paralichthys woolmani) and
Pacific yellowtail (Seriola mazatlana), new candidate species for aquaculture. Aquaculture
155, 307-318.
This paper describes the spawning and larval rearing of two new species for aquaculture
in Ecuador. Experimental production of these species has been conducted for the first time fromeggs in captivity. Survival of Seriola throughout larval rearing ranged from 0 – 70%, howeverhigh mortalities caused by diseases, cannibalism, and weaning onto artificial diets during andafter metamorphosis reduced average survival rates through the juvenile stage to less than 1%.
The basic broodstock and larval rearing information provided by the author is a good starting
point for the culture of either Paralichthys or Seriola culture.
Moran, D., C.K. Smith, B. Gara, C.W. Poortenaar. 2007. Reproductive behaviour andearly development in yellowtail kingfish (Seriola lalandi Valenciennes 1833). Aquaculture
262, 95-104.
The authors describe the spawning behaviour of wild caught brood stock as well as early
egg and larval development of yellowtail kingfish (Seriola lalandi). Spawning occurred
naturally when water temperature was above 17 C. Courtship behaviour involved one male andfemale, and consisted of a high-speed pursuit punctuated by stalling, nipping and touching. This
lasted for approximately 0.5 – 1.5 h until, immediately prior to spawning, the male would nip at
the female gonoduct, presumably to induce spawning. Spawning occurred in the early daylight
hours at the start of the spawning season, but shifted to around dusck in the latter part. Spawned
eggs were positively buoyant, had a high fertilization rate (>99%), ranged 1.33 – 1.50 mm indiameter with a single oil droplet 0.30 – 0.33 mm diameter. Information given in this paper isindicative of a healthy spawning population of S. lalandi and can be used as a reference orbaseline for culture of this species.
Papandroulakis, N., C.C. Mylona, E. Maingot, P. Divanach. 2005. First results of greateramberjack (Seriola dumerili) larval rearing in mesocosm. Aquaculture 250, 155-161.
The authors apply the mesocosm method for larval rearing to the greater amberjack(Seriola dumerili). This paper gives a good description of larval rearing for Seriola in terms of

UJNR Annotated Bibliography
weaning schedule, turbidity levels, and live feed levels. Along with the description of therearing protocols, growth information is provided which is a good template for most Seriola
species.
Sakakura, Y. and K. Tsukamoto. 1999. Ontogeny of aggressive behaviour in schools ofyellowtail, Seriola quinqueradiata. Environmental Biology of Fishes 56, 231-242
The authors studies ontogenetic changes in social interactions, especially in aggressive
behaviour of the yellowtail, Seriola quinqueradiata, and compared these to morphological andphysiological changes. No agonistic interactions were observed during the larva period until 10
mm in total length (TL) at approximately 20 days after hatching. Typical shivering behaviour
with ‘J-posture’ was observed during metamorphosis, when fin rays and calcification of vertebra
were completed and there was an increase of tissue thyroid hormone. The onset of aggressive
behaviour was just after metamorphosis to the juvenile period, and coincided with a significant
increase in tissue cortisol levels. The onset of schooling behaviour was at 12 mm TL, slightly
after the onset of aggressive behaviour. The authors provided a good basis for determining
aggressive behaviour in Seriola quinqueradiata which can be used as a model for other Seriola
species. This paper also presents possible solutions to aggressive interactions found in theculture environment.
Yoseda K., K. Yamamoto, K. Asami, M. Chimuea, K. Hashimoto, S. Kosaka. 2008.
Influence of light intensity on feeding, growth, and early survival of leopard coral grouper(Plectropomus leopardus) larvae under mass-scale rearing conditions. Aquaculture 279, 5562.
This study investigated the effect of different light intensities on feeding, growth, andsurvival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different lightintensities (0, 500, 1000, and 3000 lx) were used and larvae were kept under constant light
conditions from 0 day after hatching (DAH) to 5 DAH. The results indicate that coral grouper
larvae are visual feeders and their food intake increases with increasing light intensity and thatlight intensity is the factor affecting larval feeding, growth, and survival. The authors provide asound experimental design and interesting results which can translate to any species whoselarvae are visual predators. The comprehension of optimization of environmental rearing
conditions such as light intensity as it relates to first feeding larvae is necessary in establishingstable mass-scale rearing technology for any species.

EXPERIMENTAL STUDY ON BROODSTOCK MANAGEMENT OF BARFIN
FLOUNDER UNDER THE CONCEPT OF MINIMUM KINSHIP SELECTION
Suzuki, S

UJNR Annotated Bibliography
Nobuhiko Taniguchi. 2004. Broodstock management for stock enhancement programs ofmarine fish with assistance of DNA markers (a review). Stock Enhancement and Sea
Ranching. developments, pitfalls and opportunities (2nd Ed.). Pp 329-338.
The microsatellite DNA markers have been used, and the availability of the markers wasrecognized in some examples of studies on the genetic divergence and broodstock managementof red sea bream for enhancement programs in Japan. Based on these microsatellite DNA data,
genetic variability, effective population size and inbreeding coefficients could be estimated forbroodstocks in hatchery. A procedure for broodstock management is proposed to maintain thegenetic variability of the target fish species. If the genetic divergence is decreased in the broodstock, the minimal-kinship criterion on relatedness estimator may be applied to recover the
genetic diversity of the founder population by selective breeding based on this criterion. This
procedure might be useful not only in the case when low numbers of parental fish are available,
but also in the case of conservation of genetic variability in threatened species.

IMPROVING THE HATCHERY OUTPUT OF THE HAWAIIAN PINK SNAPPER,PRISTIPOMOIDES FILAMENTOSUS
Tamaru, C
Morote1, E., M.P. Olivar, P.M. Pankhurst, F. Villate, and I. Uriarte. 2008. Trophic
ecology of bullet tuna Auxis rochei larvae and ontogeny of feeding-related organs. Mar.
Ecol. Prog. Ser. 353: 243–254, doi: 10.3354/meps07206
This is a comprehensive study on the diet of early fish larvae of a tuna species collectedfrom the Mediterranean. Assessment of diet was done through gut content analysis in larvae
ranging from approximately 2 mm to > 5 mm. In addition, the authors analyzed the developmentof the digestive system and the eye. Both digestive capacity and visual acuity are changing
rapidly in these early larvae, and thus impact what they are capable of seeing, catching anddigesting. In terms of diet, the two important conclusions of the authors are: 1) even thesmallest larvae ingest a large variety of prey items; 2) size of prey increases with size; and 3)
feeding is selective and changes with fish size. These types of ecological studies provide a basis
for trying different food types in captivity. However, there are limitations to gut content
analysis, and these need to be considered. In particular, small and soft bodied prey may beunderestimated in guts due to their rapid digestion.
Herbing, I.H. von, S.M. Gallager, W. Halteman. 2001. Metabolic costs of pursuit andattack in early larval Atlantic cod. Mar. Ecol. Prog. Ser. 216: 201-212
This laboratory-based study provides a cost-benefit analysis for feeding in early larvae of
the Atlantic cod. Two prey items are contrasted: copepod nauplii (Pseudodiaptomus sp.) andUJNR Annotated Bibliography
protozoans (Balanion sp.) in 3 sized classes oflarvae (5-6, 6-7 and 7-8 mm). The behavioral
analysis in this study underscores the differences in
the capture of an evasive vs. a non-evasive preyusing both respiration rates and swimming speed
analyses. Smallest fish larvae were not able tocapture copepod nauplii, but fed readily on
protozoans. The main conclusion of this study isthat a smaller prey is less costly to catch, but also
provide much lower benefit. The larger prey
(copepod nauplii) required 2-3x more energy tocapture, but provided two orders of magnitude
greater benefit it terms of calories.
VanderLugt, K. ,M. J. Cooney, A. Lechner, and P. H. Lenz.. 2009. Cultivation of the
Paracalanid Copepod, Bestiolina similis (Calanoida: Crustacea). World Aquaculture
Society, Volume: 40(5):616-628.
VanderLugt, K., B. Clauberg, P. Lenz. 2005. Intensive Cultivation of the Calanoid
Copepod Bestiolina similis. Volume 12, Issue 4 on 01 April 2005.
http://www.jyi.org/research/re.php?id=147
The two references above summarizes the work done as part of a Master’s Dissertationand documented the temporal changes in production in batch cultures of the calanoid copepod,
Bestiolina similis. The focus of the research was to develop management strategies for the use of
copepod nauplii as a live food. Population abundances and female egg production rates were
measured, and recruitment and mortality rates were calculated. The copepod, B. similis fed withthree phytoplankton strains: Chaetoceros neogracille, Isochrysis galbana Tahitian strain,
Figure 1. Temporal changes in nauplii andand Rhodomonas sp. at various concentrations
resulted in highest survival rates with. I. galbana adult copepod density in response to an I.
galbana diet.
at a concentration of 5.0 x 104 cells ml-1.
However, when copepods were isolated fromeach culture every 12 hours and egg production monitored, it was also found that copepods
fed with I. galbana had the highest egg production. The highest average egg production
rate was 23 eggs female-1 day-1 with the maximum measured at 30 eggs female-1 day-1. The
most intriguing observation from a culture standpoint was that the production cycle was highly
predictable (Figure 1) and provided an opportunity to coordinate nauplius production rates with
first feed needs of fish larvae. These observations would be replicated in 3,000-L larval rearing
tank that would result in high survival during the first feeding stage of the pink snapper.
VanderLugt, K. and P.etra Lenz. 2009. Handbook for the Cultivation of two Hawaiian

UJNR Annotated Bibliography
Paracalanid Copepods. Technical Report: Pacific Biosciences Research Center. Universityof Hawaii at Mano. 19 pp.
This technical handout, that is available on the internet, is an excellent compilation of thebiology, morphology, identification and step by step description of the techniques of the culture
process for the calanoid copepod, Bestiolina similis. The descriptions include collection of
copepods from the wild and how they are sorted and processed for culture. Information also
covers the production of phytoplankton cultures needed to feed the copepods. The guides areuseful for small scale culture processes.

UNDERSTANDING ARTEMIA BIOGEOGRAPHYPRODUCTION AND JUVENILE QUALITY

RELATEDTO

HATCHERY

Torrentera, L
Seenivasan C., C., P. Saravana Bhavan, S. Radhakrishnan, R. Shanthi. 2012. Enrichmentof Artemia nauplii with Lactobacillus sporogenes for Enhancing the Survival, Growth andLevels of Biochemical Constituents in the Post Larvae of the Freshwater Prawn
Macrobrachium rosenbergii. Turkish Journal of Fisheries and Aquatic Sciences. 12: 23-3
The authors in this study designed and experiment to prove that Artemia could be used as
vector to deliver probiotics to improve larva digestion and growing. They used Artemia
franciscana naupli enriched with bacterium, Lactobacillus sporogenes and fed the freshwater
prawn, Macrobrachium rosenbergii post larvae (PL). Artemia was enriched with different
concentrations of L. sporogenes. Enriched Artemia produced significantly (P<0.05) higher
survival and growth when compared with the control fed with un-enriched Artemia. M.
rosenbergii (PL) biomass increase, total weight gain, specific growth rate increase, condition
factor and mean conversion ratio were found to be higher in PL fed with L. sporogenes enriched
Artemia when compared with control. The enhanced growth performance of PL fed with L.
sporogenes was further confirmed by the lower food conversion ratio recorded. The levels of
biochemical constituents, such as total protein, amino acids, carbohydrate and lipid contents
were found significantly higher (P<0.05) in M. rosenbergii PL fed on L. sporogenes enriched
Artemia particularly when compared to the un-richer control. The authors concluded that cells of
L. sporogenes using Artemia as a vector can be considered as suitable probiotic for attaining
good survival and growth of M. rosenbergeii post Larvae.
Soltanian Siyavash, Jean Dhont, Patrick Sorgeloos, Peter Bossier. 2007. Influence ofdifferent yeast cell-wall mutants on performance and protection against pathogenic
bacteria (Vibrio campbellii) in gnotobiotically-grown Artemia. Fish & Shellfish Immunology
23: 141-153

UJNR Annotated Bibliography

The present study determined the selection of isogenic yeast strains (with deletion forgenes involved in cell-wall synthesis) to evaluate their nutritional and immune-stimulatory
characteristics for gnotobiotically-grown Artemia. A set of experiments showed the nutritional
value of isogenic yeast strains (effected in mannoproteins, glucan, chitin and cell-wall boundprotein synthesis) for gnotobiotically-grown Artemia. Yeast cell-wall mutants were always better
feed for Artemia than the isogenic wild type (WT) mainly because they supported a higher
survival. The difference in Artemia performance between WT and mutants feeding was reduced
when stationary-phase grown cells were used. These results suggest that any mutation affecting
the yeast cell wall make-up is sufficient to improve the digestibility in Artemia. The second setof experiments, investigates the use of a small amount of yeast cells in gnotobiotic Artemia toovercome pathogenicity of Vibrio campbellii (VC). Among all yeast cell strains used in thisstudy, only mnn9 mutant yeast (less cell-wall bound mannoproteins and more glucan and chitin)
seems to completely protect Artemia against V C. The result with these mutants is of particularinterest, as its nutritional value for Artemia as comparable to the wild type. The results of thisstudy suggest non-interference of Artemia general nutritional effects.
Shadrin Nickolai, Elena Anufriieva, Ekaterina Galagovets. 2012. Distribution andhistorical biogeography of Artemia leach, 1819 (Crustacea: Anostraca) in Ukraine.
International Journal of Artemia Biology. Vol. 2, No 2: 30-42
In this study the authors described the recent information about the biogeography of
Artemia in Ukraine (Ukraine separated from the Soviet Union after the cold war). The authors
identified the following areas with potential for aquaculture: The Crimean peninsula; the coastalzone of the NW Black Sea; the north coast of the Sea of Azov; Donetsk Oblast; Zakarpattia
Oblast. These authors recognized that at least there are two bisexual species (A. salina and A.
urmiana) and the parthenogenetic complex populations of Artemia dwell in the Ukraine. In somelakes (Koyashskoye, Terecly-Konradskoye) they found A. urmiana and parthenogens both, but
separated in time. In the waters of the Crimea and NW part of the Black Sea brine shrimp arefound under salinities from 10-20 ppt (if no other animals present) to 350 - 370 ppt. Larvae andadults of some Coleoptera, Hemiptera, ostracods Eucypris inflata (Sars) were found, andprobably other invertebrates may eat away Artemia completely in the Crimean lakes whosesalinity is below 60-110 ppt. Exploitation of salt can create new habitat for brine shrimp. Ukraine
hypersaline ecosystems are very promising opportunities for industrial harvesting of Artemia
cysts.
Cohen Rosa Graciela. 2012. Review of the biogeography of Artemia Leach, 1819
(Crustacea: Anostraca) in Argentina. International Journal of Artemia Biology Vol. 2, No 1:
9-23.

UJNR Annotated Bibliography
In this study the author make a review of the biogeography of Artemia in Argentina andother countries in South America. She describes two species of Artemia in these countries: A.
franciscana widely distributed throughout the Americas and the Caribbean, and A. persimilis
with a more restricted distribution in southern South America (Argentina, Chile, Peru, andBolivia). In Argentina A. franciscana is located in the Buenos Aires Province. A. persimilis,
from: La Pampa and Santa Cruz Provinces. The southernmost record of the species corresponds
to Laguna de los Cisnes in the Chilean Tierra del Fuego. Temperature is considered as a main
factor in the determination of the present geographic distribution of both species in thesecountries. A. franciscana and A. persimilis are considered as endemic species to the Americas.
However, in recent decades, A. franciscana was intentionally introduced in several countries of
the New and the Old World, mainly associated with aquaculture. Two African populations
recently were assigned as A. franciscana suggesting other natural events raising the hypothesisthat the original geographic distribution of this species was not only restricted to the Americas.
On the other hand, A. persimilis was described for the first time in Salinas Grandes de Hidalgo,
Argentina and in Europe from the Italian population of Saline di San Bartolomeo, Sardinia. A.
persimilis (now extinct) co-occurred with A. salina in the later location. The presence of several
Artemia persimilis populations at high-altitude (above 4,500 m) in the South America Altiplano
could lead to an hypothesis that suggest that in the geological past, A. persimilis would haveretreated to cooler and higher altitudes and latitudes, leaving isolated populations in Andean
enclaves.
Torrentera Laura and Stanley I. Dodson. 2004 Ecology of the brine shrimp Artemia in the
Yucatan, Mexico, Salterns. Journal of Plankton Research. Vol 26:(5) 1-8.
In this article the authors described the biogeography and ecology of the brine shrimp
Artemia across the Yucatan peninsula. They recorded monthly field data during one year at theNorth Coast hypersaline ecosystems: temporary pond, pools, salterns and saltmarshes. The
Yucatan Peninsula is a great limestone platform projecting from Mexico and Central America
northward into the Gulf of Mexico and the Caribbean Sea. The most important ports sampledincluded: Celestun, at the extreme north west of the peninsula and Chuburna and, Xtampu, at theintermediate area and, Las Coloradas, at the extreme northeastern part of the peninsula. The
salinity ranges from that of seawater (35 g/L) to about 150 g/L or more. The locations revealed
high extreme alkalinity, salinity and temperature conditions, and hypoxia and in some instances,
total desiccation. The four locations were similar in water temperature, nitrites, and phosphates,
but differed in salinity, oxygen content, water depth, nitrates, ammonia, silicates, carbonates,
sulfates, and pH. The dominant multicellular organism in these habitats is Artemia, the brine
shrimp. Artemia abundance and population dynamics were significantly correlated with specificenvironmental conditions. The annual natural production mean value of Biomass calculated in
the sampled area was 450 D. W. (g/square meter). The cyst production mean D. W. value was
733 (g/square meters). The field study provides ecological basis for the exploitation of Artemia
as a live food that possess good nutritional value for larvae culture of finfish and crustaceans.

UJNR Annotated Bibliography
These hypersaline ecosystems should be protected because they are nesting and feeding areas for
the Caribbean flamingo and other marine birds.

CONDITIONING TECHNOLOGIES FOR FLATFISH STOCK ENHANCEMENT:GLOBAL PROGRESS AND PITFALLSWalsh, M
Walsh, M. L., E. A. Fairchild, N. Rennels, S. C. Farina, W. H. Howell, R. Mercaldo-Allen,
and C. Kuropat. 2009. Rearing Diets for Winter Flounder Optimize Weaning Success in
Hatchery, Wild. Global Aquaculture Advocate. May/June: 48-50.
The objectives of this work were to quantify feeding-related performance of reared-thenreleased winter flounder juveniles that were reared on different feeds (both live and formulated)in the hatchery. The authors evaluated how feeding history translated to wild feeding
performance once individuals were released into nature (caged in-situ) by examining survival,
growth, feeding onset and incidence, stomach fullness, diet composition, and nucleic acid-based
condition. Fish raised on live diets exhibited higher survival and growth than those reared on
formulated, pellet feeds.
Støttrup, J. and C. R. Sparrevohn. 2010. Stock enhancement Europe: Turbot Psetta
maxima, pp. 219–236., in: Practical Flatfish Culture and Stock Enhancement (H.
V. Daniels and W. O. Watanabe, Eds.) Ames, IA, USA: Wiley-Blackwell
Publishing.
This chapter provides a thorough yet concise description of Atlantic turbot releases
throughout Europe. The authors discuss the aims and rationale of turbot stocking and include
information regarding marking and tagging techniques, choice of release site, release strategies,
and conditioning.
Walsh, M. L., H. Fujimoto, T. Yamamoto, T. Yamada, Y. Takahashi, and Y. Yamashita.
2012. Case Studies in Flatfish Stock Enhancement: A Multi-Year Collaborative Effort to
Evaluate the Impact of Acclimation Cage Conditioning for Japanese Flounder,
Paralichthys olivaceus, in Wakasa Bay, Japan. US-Japan National Resources Panel on
Aquaculture (UJNR) 39th Proceedings. Kagoshima, Japan. 35: 93-102.
The authors examined both the immediate (burying, feeding) and longer-term(movements, recapture rates) benefits of cage conditioning juvenile Japanese flounder before
stocking. Recapture rates of cage-conditioned fish caught by local fishermen were greater thanthose of non-conditioned fish (i.e., those released directly from hatchery tanks). In addition,UJNR Annotated Bibliographyburying ability was assessed by releasing recently conditioned and non-conditioned fish into
tanks and quantifying the number of fish that buried. Likewise, feeding ability was assessed byproviding tanks of conditioned and non-conditioned fish with prey and quantifying how many
prey remained after a period. These laboratory experiments revealed that conditioned fishexhibited enhanced burying and feeding performance compared to non-conditioned fish. This
work was the first to examine flatfish conditioning strategies using market data and to evaluate
experimentally the performance of hatchery-reared flatfish that have been cage-conditioned.
Fairchild, E. A., N. Rennels, and W. H. Howell. 2008. Predators are attracted toacclimation cages used for winter flounder stock enhancement. Rev. Fish. Sci. 16, 262–168.
This paper addresses a negative consequence of using acclimation cages for stocking fish:that the cages in themselves may attract predators. Crab densities significantly increased in thevicinity of acclimation cages and continued to increase each day, even when cages were empty(i.e., contained no fish). Cages containing fish attracted an even higher abundance of crabs.

CHARACTERISTIC EVALUATION METHOD RELATINGDISEASE OF YELLOWTAIL (SERIOLA QUINQUERADIATA)TO

BENEDENIA

Yoshida, K
Mushiake, K., H. Yamazaki, and H. Fujimoto. 2005. Current situation of technicaldevelopments in seed production of yellowtail (Seriola quinqueradiata) in japan.
Proceedings of the thirty-fourth U.S.-Japan aquaculture panel symposium (eds R.
Stickney, R. Iwamoto, and M.Rust ) : Pp1-4.
The National Center for Stock Enhancement (NCSE, formerly Japan sea-farming
Association), of the Fisheries Research agency, introduced the stock enhancement program foryellowtail (Seriola quinqueradiata and Seriola lalandi) in 1977. Technical developments in
induced spawning as well as larval and juvenile rearing techniques have increased the populationof this species to 1 million juveniles per year at NCSE. This project faced three major
drawbacks: high mortality of larvae, cannibalism, and the smaller size of released juveniles incomparison with their wild counterparts. The high mortality of larvae was overcome by utilizing
strong aeration during the early larval stage, while cannibalism was controlled by grading
juveniles by size selection. The two-month delay in the spawning season of reared broodstock(the usual spawning season is late April to early May), which caused the smaller size of released
juveniles, was solved by developments in advanced spawning techniques. Photoperiod and watertemperature manipulations were used toproduce eggs in February, thus producing yellowtail
juveniles that can be released into the wild at a size similar to that of the wild stock.

UJNR Annotated Bibliography
Ohara, E., T. Nishimura, Y. Nagakura, T. Sakamoto, K. Mushiake, and N. Okamoto. 2005.
Genetic linkage maps of two yellowtails (Seriola quinqueradiata and Seriola lalandi).
Aquaculture 244 : Pp41-48.
The yellowtails Seriola quinqueradiata and Seriola lalandi are the most important
species in marine fishery resources and aquaculture in Japan. A genetic linkage map is needed to
improve efficiency of breeding by marker-assisted selection (MAS) and for the identification ofcommercially important genes. Therefore, we have constructed a genetic linkage map for theyellowtails using microsatellite makers. Microsatellites were isolated from agenomic DNA
library of S. quinqueradiata. Segregation of 217 microsatellites was studied in 90 progeny from across between S. quinqueradiata and S. lalandi. These were used to construct separate linkage
maps of a female (S. quinqueradiata) and a male (S. lalandi). Twenty-five linkage groups weredistinguished in the female (S. quinqueradiata) map, which spanned for 473.3 centiMorgans(cM) with an average intermaker distance of 2.7 cM (total length estimated: 901.7 cM), and 21
linkage groups were distinguished in the male (S. lalandi) map, which spanned 584.3 cM with anaverage intermaker distance of 4.8 cM (total length estimated: 1715.3 cM). The microsatellite
loci and genetic linkage maps will increase the efficiency of selective breeding programs foryellowtails.
Nagakura. Y., T. Yoshinaga, T. Sakamoto, K. Hattori, and N. Okamoto (2010).
Susceptibility of four families derived from two Seriola species to the monogenean parasite(Benedenia seriolae) using a new challenge method. Journal of Fisheries Technology 3:
Pp21–26 (in Japanese)
Four families of two Seriola species were experimentally challenged with oncomiracidia
of Benedenia seriolae. One family was produced by crossing wild yellowtails (S.
quingueradaiata), two families were crosses between yellowtails of the 4th or 5th generationpassaged under laboratory conditions, and one hybrid family was a gross between yellowtail andgoldstriped amberjack (S. lalandi). Four hundred juveniles, 100 per each of the four groups andranging 108-111 mm in mean total length, were exposed to about 40,000 oncomitacidia of B.
seriolae within 30 min after hatching in a 1000L tank in darkness for 4 hours. On day 10 postchallenge, the number and size of parasites were estimated on each fish. Infection intensities
significantly differed in any combination of families. A significant difference was also detectedin worm body length in a combination of two families. The results suggest that families of fish
resistant to B. seriolae can be produced through breeding. The advantages of the method
employed in this study for evaluating fish susceptibility against B. seriolae are also discussed.