Aquaculture Nutrition

2016 22; 25–33

..........................................................................................
doi: 10.1111/anu.12228

1 1 1,2 3 1
1
Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo,
Japan; 2 Faculty of Life Sciences, Toyo University, Itakura, Oura, Gunma, Japan; 3 Faculty of Nutritional Science,
Sagami Women’s University, Sagamihara, Kanagawa, Japan

Phyllosomas (planktonic larvae) of slipper lobsters cling Mass production of juveniles in spiny (palinurid) and slip-
onto and feed on jellyfish under both natural and labora- per (scyllarid) lobsters is a critical step for the commercial
tory conditions. Phyllosomas of Ibacus novemdentatus are production and aquaculture of these species. One of the
capable of feeding on various jellyfish species including principal issues hampering successful mass production of
venomous stingers; however, the range of jellyfish species juvenile lobsters is the limited information available on
capable of supporting the growth and survival of phylloso- suitable dietary regimes for growth and survival during the
mas is unknown. Seventeen (12 for the first and five for the long phase of planktonic larvae (phyllosomas) (Cox &
second trials) and 18 (13 for the first and five for the sec- Johnston 2003; Cox et al. 2011). To this end, many
ond trials) phyllosomas were fed exclusively on the jellyfish researchers have focused on an understanding of phylloso-
Aurelia aurita and Chrysaora pacifica, respectively. Aurelia ma development. The complete development of phylloso-
aurita-fed phyllosomas metamorphosed into the nisto stage mas from hatching to settlement has been reported in 10
(postbenthic larvae)  54 days after hatching, whereas (Phillips & Matsuda 2011) of 32 (Groeneveld et al. 2013;
C. pacifica-fed phyllosomas did not. Major nutritional Jeffs et al. 2013; Phillips et al. 2013) and 11 (Vijayakuma-
compositions such as amino acids, fatty acids and minerals ran & Radhakrishnan 2011) of 33 (Spanier & Lavalli 2013)
were compared between the two jellyfish species. The pro- economically important species of spiny and slipper lob-
portion of each major nutritional component was not sig- sters, respectively. However, the mass production of juve-
nificantly different between the two jellyfish species, nile lobsters has only been achieved in a slipper lobster
suggesting that C. pacifica was not nutritionally inferior to Thenus orientalis (Mikami & Kuballa 2007).
A. aurita. Therefore, the observation that the C. pacifica- There is abundant evidence that wild phyllosomas associ-
fed phyllosomas did not metamorphose into the nisto ate with and feed on jellyfish (reviewed by Booth et al.
stage was not because of major nutritional compositions 2005; Sekiguchi et al. 2007; Wakabayashi & Tanaka 2012).
but due to other factors such as the lack or excess of other For example, phyllosomas of the genus Ibacus and Scylla-
minor nutrients, or the species-specific texture of the rus are known to cling usually onto the cnidarian jellyfish
jellyfish. (e.g. Herrnkind et al. 1976; Ates et al. 2007), and those of
Parribacus antarctics collected from the north of Bermuda
KEY WORDS: amino acids, fatty acids, growth, lobster, minerals, possessed several types of cnidarian nematocysts in their
phyllosoma faecal matter (Sims & Brown 1968). In addition, those of
Panulirus japonicus and Scyllarides squammosus collected
Received 7 January 2014; accepted 19 June 2014 from Atlantic and Pacific waters utilized cnidarians and
Correspondence: K. Wakabayashi, Tokyo University of Marine Science
urochordates as food, according to the DNA analysis of
and Technology, 4–5–7 Konan, Minato, Tokyo 108–8477, Japan. their gut contents (Suzuki et al. 2006). Laboratory experi-
E-mail: [email protected] ments also suggest that gelatinous zooplankton is a suitable
diet for phyllosomas (e.g. Goldstein & Nelson 2011). In a
..............................................................................................

ª 2015 John Wiley & Sons Ltd

series of prey feeding trials, Saunders et al. (2012) found versity of Marine Science and Technology (TUMSAT). Jel-
that phyllosomas of Panulirus cygnus preferred chaetogn- lyfish bell diameters ranged 100–260 mm for A. aurita and
aths and salps to krill, and Wakabayashi et al. (2012b) 80–200 mm for C. pacifica. All the jellyfish were transported
found that those of Ibacus novemdentatus showed a higher to the laboratory immediately after collection. In the same
survival rate when fed on jellyfish rather than Manila clam, method described by Purcell et al. (2013), the jellyfish for
Ruditapes philippinarum. Morphological analyses have also phyllosomal feeding trials were maintained in a 500-L tank
suggested that mouthparts and internal organs of phylloso- (density of one individual per 5 L) with recirculating sea
mas are adapted for masticating and digesting the soft tis- water without feeding. For biochemical analyses, the jelly-
sues of gelatinous zooplankton (Nishida et al. 1990; Cox fish were washed with freshwater immediately after trans-
& Johnston 2003; Simon et al. 2012). port to the laboratory and freeze-dried under vacuum
The next major step towards the mass production of lob- pressure (10–20 °C and 5 9 100–5 9 10 1 Pa) using a freeze
ster juveniles will be the identification of essential dietary dryer (VD-800R; Taitec Co., Tokyo, Japan).
requirements that influence the growth and survival of
phyllosomas (Cox & Johnston 2003). A key element is the
study of the biochemical compositions of potential feed-
stock (Jones et al. 1997; Wang & Jeffs 2013). All gelatinous Ibacus novemdentatus was chosen for this study because
zooplankton including jellyfish may not always be suitable phyllosomas of this species were known to feed on various
for the growth and survival of phyllosomas because their jellyfish species (Wakabayashi et al. 2012a). A total of four
biochemical compositions vary depending on the species, egg-bearing females were caught off Kitaura, Miyazaki Pre-
collection dates and locality (Lucas 1994; Nelson et al. fecture, on April 5, 2012, and transported to the laboratory
2000; Wang & Jeffs 2013). Kittaka (2005) suggested that of TUMSAT. The females were maintained at 23 °C with
the growth and survival of phyllosomas were affected by recirculating sea water until phyllosoma hatching. We
differences in the nutritional value of dietary jellyfish made duplicate feeding trials in this study (Table 1). The
although dietary regimes used in that study were altered phyllosomas originated from two females that released
during the course of the feeding cycle. Here, we examined thousands of phyllosomas on April 15 and 19, 2012,
the growth and survival of phyllosomas fed exclusively on respectively. A day after hatching, a total of 25 and 10
the jellyfish Aurelia aurita or Chrysaora pacifica (both are phyllosomas, which were swimming near the water surface
scyphozoans) to clarify the effect of different dietary jelly- with intact appendages, were chosen as specimens for the
fish species on phyllosoma development. The major nutri- respective trials.
ents in both jellyfish species were determined, and the
impact of the nutritional values of these jellyfish on the
growth and survival of phyllosomas is discussed.
The newly hatched phyllosomas were kept individually in
PVC mesh cases (width, 70 mm; length, 70 mm; and
height, 140 mm) and placed on a perforated plate 50 mm
from the bottom of the tank (width, 600 mm; length,
300 mm; and height, 360 mm). The tank contained approx-
Aurelia aurita and C. pacifica were captured using scoop imately 30 L of sea water and was equipped with an exter-
nets from Tokyo Bay from April to August, 2012, on the nal filter (Eheim 2234 ecco comfort; Eheim GmbH & Co.
training ships Seiyo-maru and Hiyodori of the Tokyo Uni- KG, Deizisau, Germany) to clean water and provide a

Table 1 Numbers of Ibacus novemdentatus phyllosomas used for each experiment

No. of phyllosomas
No. of phyllosomas used for experiments metamorphosed successfully

Trial Date of hatching Aurelia aurita Chrysaora pacifica A. aurita C. pacifica

1 15 April 2012 12 13 4 0
2 19 April 2012 5 5 2 0

..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
 gentle flow (50 mL s 1). Approximately 10 L of sea water
was exchanged weekly; water temperature and salinity were
kept at 23 °C and 34 gL 1, respectively. The phyllosomas Fatty acid composition was determined by gas chromatog-
were reared under natural laboratory light conditions. raphy as described by the AOCS official method Ce 1b-89
Phyllosomas were divided into two experimental groups: (AOCS 1998) with minor modifications as follows. Capric
12 for A. aurita and 13 for C. pacifica from the April 15 acid methyl ester was added as an internal standard to
batch and 5 for A. aurita and 5 for C. pacifica from the accurately weigh total lipid in approximately 1 g of freeze-
April 19 batch (Table 1). They were fed exclusively with dried jellyfish sample. The mixture was saponified with
A. aurita or C. pacifica. Immediately before feeding, both 1.25 M HCl in methanol and methylated at 100 °C for 3 h
jellyfish species were cut into pieces approximately twice of to obtain fatty acid methyl esters (FAMEs). The FAMEs
the carapace weight of the phyllosomas. Eighteen sets of obtained were dissolved in n-hexane (Wako Pure Chemical
two mesh cases joined together, one in which the phylloso- Industries) and dehydrated with sodium sulfate (Wako
ma was fed exclusively on A. aurita or the other which the Pure Chemical Industries). The dehydrated FAMEs were
phyllosoma was fed exclusively on C. pacifica, were ran- analysed using a gas chromatograph (TRACE GC Ultra;
domly arranged in the tank to minimize variations in Thermo Fisher Scientific Inc.) equipped with a mass spec-
experimental conditions. Instars and number of surviving trometer (DSQ II; Thermo Fisher Scientific Inc.), an auto-
phyllosomas were recorded daily. sampler (AS-3000; Thermo Fisher Scientific Inc.) and a
Supelcowax 10 fused silica capillary column
(30 m 9 0.25 mm i.d. 9 0.25 lm film thickness; Sigma-
Aldrich Co., St. Louis, MO, USA). Helium gas was used
The improved Dumas method (Anderson & Shubin 1957) as a carrier at a constant flow rate of 20 mL min 1. The
was used to determine total carbon and nitrogen in jellyfish injector temperature was 250 °C. The column oven temper-
using gas chromatography (SUMIGRAPH NC-220; Sumi- ature programme was as follows: an initial temperature of
ka Chemical Analysis Service, Osaka, Japan). A calibration 50 °C for 5 min, followed by an increase from 50 to
curve was obtained with aspartic acid (Sumika Chemical 250 °C at a rate of 10 °C min 1. The final hold time was
Analysis Service). Crude protein was calculated using the 30 min. Fatty acids were identified by comparing their
nitrogen conversion factor of 6.25 (AOAC 1995), total lipid retention times with those of authentic standard com-
was determined as total fatty acids, and ash was calculated pounds (Fatty acids and their methyl esters Kit; GL Sci-
by the difference between initial and final weight of the ence, Tokyo, Japan).
samples.

For mineral analysis, microwave digestion of the freeze-

Total amino acid composition was determined according to dried samples was performed under nitrogen (Wako Pure
the method of Ogushi & Harada (1997) with minor modifi- Chemical Industries) using the ETHOS-1 microwave diges-
cations as follows. Samples were hydrolysed using HCl tion system (Milestone Inc., Shelton, CT, USA). Levels of
(3 mg freeze-dried jellyfish sample in 1 mL of 6 M HCl Ca, Mg, Na and K were determined by atomic absorption
with 0.02% of 2-methylmercaptoethanol at 110 °C for photometry (ANA-182; Tokyo Koden, Tokorozawa,
24 h) in vacuum hydrolysis tubes (Thermo Fisher Scientific Japan), and those of B, Al, Mn, Fe, Cu, Zn, As, Mo, Cd,
Inc., Waltham, MA, USA). After evaporation to remove Hg, Pb, V and Cr were determined by ICP-MS (X series 2;
HCl, protein hydrolysates were dissolved in 1–2 mL of Thermo Fisher Scientific Inc.).
0.02 N HCl and purified using DISMIC 0.45 lm filters
(DISMIC-13CP series; Advantec MFS, Inc., Dublin, CA,
USA). The hydrolysate solution was analysed using an
amino acid analyzer (L-8900; Hitachi High-Technologies Significant differences of larval duration between A. aurita-
Co., Tokyo, Japan). Amino acids mixture standard solu- fed and C. pacifica-fed phyllosomas were determined using
tion, Type H (Wako Pure Chemical Industries, Ltd., Student’s t-test. Survival rates were calculated using the
Osaka, Japan), was used for making a calibration curve to Kaplan-Meier method (Kaplan & Meier 1958). The log-
identify and quantify amino acids. rank test was applied to analyse difference of survival rates
..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
between A. aurita-fed phyllosomas and C. pacifica-fed A. aurita metamorphosed into nistos by the 54th day after
phyllosomas. Only the results of first trial were used for the hatching; the shortest metamorphosis time was 41 days
analysis because the sample number for second trial was (Fig. 1). On the other hand, no nistos were obtained from
small. the C. pacifica-fed phyllosomas (Table 2). The survival rate
For the analysis of biochemical compositions, all samples of C. pacifica-fed phyllosomas was not significantly differ-
were analysed in triplicate. Data are presented as ent from that of A. aurita-fed phyllosomas until the 25th
mean  standard deviation (n = 3). Significant differences day when most of the phyllosomas were in the fifth instar
(P < 0.05) were determined using Student’s two-sided (P = 0.91, Fig. 1). Nevertheless, after the 30th day, the sur-
t-test. vival rate in C. pacifica-fed phyllosoma dropped signifi-
cantly (P < 0.05, Fig. 1) because of the failure of moulting
from the fifth to the sixth (final) instar. Overall, growth of
the C. pacifica-fed phyllosomas gradually slowed after the
third instar. The duration of the moulting interval tended
to be longer in C. pacifica-fed phyllosomas than that in
Four phyllosomas (33.3%) from the April 15 batch and A. aurita-fed phyllosomas although no significant differ-
two phyllosomas (40.0%) from the April 19 batch fed with ences were detected except the duration from the second
instar to the third instar (Table 2). Four C. pacifica-fed
phyllosomas reached the sixth instar, and one of them
attempted to metamorphose into nisto after 55 days of
hatching; however, the attempt failed.

The quantities of crude protein, total lipid, total carbon

and total nitrogen were higher in C. pacifica than in A. au-
rita, whereas that of ash was significantly lower in C. pacif-
ica than in A. aurita (Table 3).
The quantity of total amino acids was significantly
higher in C. pacifica than in A. aurita. In addition,
Figure 1 Percentage survival of Ibacus novemdentatus phyllosomas C. pacifica showed significantly higher levels of individual
fed exclusively on Aurelia aurita (single line) or Chrysaora pacifica
amino acids than A. aurita except for phenylalanine
(double line). Each percentage is the mean of two experiments.
Arrow and grey bar indicate the average and range of time when
(Appendix 1). Although the quantities of each amino acid
metamorphosis took place, respectively. No C. pacifica-fed phyllos- were higher in C. pacifica, the percentage composition of
omas underwent metamorphosis, which is why there are no arrows them were similar between the two jellyfish species (Fig. 2,
given here.
Table 3 Proximate composition of the jellyfish species, Aurelia au-
rita and Chrysaora pacifica (mg gDW 1; mean  standard devia-
Table 2 Average duration (in days) from hatching to the achieve-
tion; n = 3)
ment in each instar in Ibacus novemdentatus phyllosoma fed exclu-
sively on Aurelia aurita or Chrysaora pacifica Jellyfish

Instar A. aurita C. pacifica Chemical parameters A. aurita C. pacifica

2 5.9  1.2 6.6  1.6 Crude protein 34.87  b

2.83 75.31  19.96a
3 10.5  1.7a 12.4  2.4b Total lipids 4.26  0.88b 7.15  1.94a
4 15.8  2.0 16.9  2.4 Ash 761.9  9.96a 690.5  9.17b
5 21.4  3.2 24.7  4.3 Carbohydrates 199.0  11.86b 227.1  11.44a
6 30.7  5.4 36.3  6.7 Total carbon 20.47  1.83b 42.28  10.93a
Nisto 47.7  4.4 N.A. Total nitrogen 5.58  0.45b 12.05  3.19a
Amino nitrogen 5.19  0.25b 9.82  0.39a
No nistos were obtained from C. pacifica-fed phyllosomas.
Different superscript letters indicate a significant difference Different superscript letters on each line indicate a significant
(P < 0.05). The duration of the third instar was significantly dif- difference (P < 0.05). Carbohydrates correspond to the organic
fered between A. aurita and C. pacifica. matter minus crude protein, total lipids and ash.
..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
 Figure 2 Average percentage of total amino acid composition of the jellyfish species Aurelia aurita (grey bars) and Chrysaora pacifica
(dotted bars) (n = 3). Asterisks (*) indicate significant differences (P < 0.05). Essential amino acids (EAA) for crustaceans are underlined.
EAA shows the sum of the nine kinds of EAA.

Appendix 1). For both species, the major amino acid was between the two species (Appendix 3). In addition, no sig-
glycine, which accounted for approximately 15% of total nificant differences were observed in the quantities of trace
amino acids, followed by glutamine and proline. Essential minerals.
amino acids (EAA) seemed to be higher in A. aurita than
in C. pacifica although no significant differences were
detected (Fig. 2, Appendix 1). More than 90% of the total
nitrogen was amino nitrogen in A. aurita, whereas 81.5% The quantities and proportion of prey and its biochemical
in C. pacifica (Table 3). composition are major factors in the selection of prey by
Among five essential fatty acids (EFA), the quantities of predators (Schoener 1971; Wang & Jeffs 2013). Lipids are
C20:4 and C22:6 were significantly higher in C. pacifica especially important nutritional components for phylloso-
than in A. aurita, whereas that of C20:5 was higher in mas since the accumulation and storage of such energy
A. aurita than in C. pacifica (Fig. 3, Appendix 2). On the constituents are critically linked to the non-feeding nisto or
other hand, the quantities of C18:2, C18:3 and the total puerulus stage (Jeffs et al. 2001). Although proteins and
EFA were not significantly different between the two spe- lipids were more abundant in C. pacifica than in A. aurita,
cies. No significant differences were detected between the I. novemdentatus phyllosomas were able to survive by
quantities of polyunsaturated, monounsaturated and satu- exclusive feeding on A. aurita as well as C. pacifica. Indeed,
rated fatty acids. Moreover, the percentage composition of we confirmed that the proportion of major nutritional com-
each fatty acid was similar between the two species (Fig. 3, ponents was basically similar between the two species,
Appendix 2). which means both A. aurita-fed and C. pacifica-fed phyllos-
Although the quantities of ash and major minerals were omas could utilize the similar major nutritional compo-
significantly lower in C. pacifica than in A. aurita, the nents for their growth and survival. We also confirmed
percentage composition of each major mineral was similar that both A. aurita and C. pacifica contained amino acids

Figure 3 Average percentage of total fatty acid composition of the jellyfish species Aurelia aurita (grey bars) and Chrysaora pacifica (dotted
bars) (n = 3) Asterisks (*) indicate significant differences (P < 0.05). Others include C11:0, C12:0, C17:1, C18:4, C19:0, C20:0, C20:1, C20:3
and C22:1. C18:2 and C18:3 are shown here although the levels of both were < 2% of the total. Essential fatty acids (EFA) for crustaceans
are underlined, and EFA shows the sum of the five kinds of EFA. Polyunsaturated fatty acids (PUFA), monounsaturated fatty acids
(MUFA) and saturated fatty acids (SFA) also show the sum of PUFA, MUFA and SFA, respectively.
..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
and fatty acids essential for crustacean nutrition, support- tors in each instar of phyllosomas should be examined in
ing previous studies of A. aurita (Fukuda & Naganuma the future.
2001; Kittaka 2005). Moreover, six essential minerals (Ca, Similar results have been reported for two species of
Cu, K, Mg, Se and Zn), which must be supplied in the diet mussel, Mytilus coruscus and M. galloprovincialis, both
for crustaceans (Davis & Gatlin 1996), were found in both considered suitable diets for P. japonicus phyllosomas
jellyfish species. Phosphate, also essential for crustaceans, (Takeuchi 2010; Murakami et al. 2011). Mytilus gallopro-
had previously been found in both A. aurita and C. pacif- vincialis-fed phyllosomas in the final instar could com-
ica (Fukushi et al. 2004). All of the nutritional data are pletely metamorphose into puerulus but not M. coruscus-
presented as dry weight, and no moisture content data are fed phyllosomas. There were few differences in biochemical
provided here; however, we here assume that moisture con- composition between the two species (Takeuchi 2010).
tent of both jellyfish species was not different, approxi- Takeuchi (2010) suggested that the most important charac-
mately 96% as reported by Fukushi et al. (2004). These teristic of the diet was the texture rather than the biochemi-
results suggest that a diet containing larger quantities of cal composition. Similarly, the differences in the texture
major nutritional components is not always suitable for between the jellyfish species used in this study may have
growth and survival of phyllosoma. affected phyllosoma feeding (e.g. mastication of diets and
Phyllosomas fed exclusively on C. pacifica could not absorption of nutrients). Further studies on the relationship
complete metamorphosis into the nisto stage, whereas between food texture and phyllosoma development are
those that fed on A. aurita could. The same result was needed to clarify this issue.
obtained from a duplicate experiment. Although the num- Our results do not rule out C. pacifica as a dietary sup-
ber of tested phyllosomas was small, it may not be inap- plement for phyllosoma. We have previously reported that
propriate to conclude that A. aurita is better than I. novemdentatus phyllosomas could metamorphose into
C. pacifica as food of the phyllosomas. Survival rate of nisto if they fed on both C. pacifica and A. aurita (Waka-
the A. aurita-fed phyllosomas was similar to that bayashi et al. 2012b). Considering their ability to prey not
observed in our previous experiments (Wakabayashi et al. only on jellyfish but also on Manila clams (Takahashi &
2012b) as well as the negative effects on the survival of Saisho 1978), I. novemdentatus phyllosoma appear to be an
the C. pacifica-fed phyllosomas (data not shown). Consid- opportunistic feeder as reported in phyllosomas of other
ering the failure of moulting and the rapid decrease of species (Jeffs et al. 2004). In the natural environment, phyl-
survival rate only in the late instars of C. pacifica-fed losomas are teleplanic, and this extended planktonic phase
phyllosomas, a minor difference between A. aurita and may benefit from feeding on a wide variety of zooplankton
C. pacifica may have critically influenced the growth and in the open sea where prey organisms are scarce (Jeffs et al.
survival of the late instar phyllosomas. We did not ana- 2004; Sekiguchi et al. 2007; Chow et al. 2011). However,
lyse minor nutrients such as phospholipids, sterol and this is not a suitable strategy for commercial production of
vitamins, which are all important for crustaceans (Kanaz- juvenile lobsters. These results suggested that the diets nec-
awa 2000), nor the clusters of related nutritional ele- essary for phyllosomas to maintain high growth rates and
ments. These minor but essential elements might affect survival and to complete metamorphosis were specific and
less for the survival of the late instar phyllosomas in that A. aurita was a suitable diet for I. novemdentatus phyl-
C. pacifica than in A. aurita. It is also possible that a losomas.
sublethal factor was accumulated in phyllosomas by feed-
ing jellyfish. Heavy metals (such as Cd, Pb, As and Hg)
included in food can have acute toxicity to lobsters
(Bryan 1971; Canli & Furness 1995), and crustacean lar- The authors thank Dr. Hiroshi Nagai, Dr. Haruto Ishii,
vae are considered to be greatly sensitive to metal toxicity and the captains and crew members of the Seiyo-maru and
(Ahsanullah & Arnott 1978). Although the composition Hiyodori, Tokyo University of Marine Science and Tech-
of heavy metals was similar between both jellyfish species, nology (TUMSAT), for collecting jellyfish. We also thank
accumulation rates of these metals into phyllosomas were Dr. Seiji Nagasaka, Toyo University, for his kind help with
not examined in this study. Consumption rates between analysing mineral composition. This study was supported
A. aurita-fed and C. pacifica-fed phyllosomas should be by Grants-in-Aid for JSPS Fellows to K.W. (No.
compared, and quantitative analyses of minor nutrients in 25•10983), the Programme for Promotion of Basic and
these two jellyfish species and accumulated sublethal fac- Applied Researches for Innovations in Bio-oriented
..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
 Industry to Y.T., and Research Promotion for Marine Sci- the continental shelf of New Zealand. Comp. Biochem. Physiol.
ence and Technology in TUMSAT to Y.T. A Mol. Integr. Physiol., 129, 305–311.
Jeffs, A.G., Nichols, P.D., Mooney, B.D., Phillips, K.L. & Phleger,
C.F. (2004) Identifying potential prey of the pelagic larvae of the
spiny lobster Jasus edwardsii using signature lipids. Comp. Bio-
chem. Physiol. B Biochem. Mol. Biol., 137, 487–507.
Ahsanullah, M. & Arnott, G.H. (1978) Acute toxicity of copper, Jeffs, A.G., Gardner, C. & Cockcroft, A. (2013) Jasus and Sag-
cadmium, and zinc to larvae of the crab Paragrapus quadridenta- mariasus species. In: Lobsters: Biology, Management, Aquacul-
tus (H. Milne Edwards), and implications for water quality crite- ture and Fisheries, 2nd edn (Phillips, B.F. ed.), pp. 259–288.
ria. Aust. J. Mar. Freshw. Res., 29, 1–8. Wiley-Blackwell, West Sussex, UK.
American Oil Chemists’ Society (1998) Official method for marine Jones, D.A., Yule, A.B. & Holland, D.L. (1997) Larval nutrition.
oil fatty acid composition by GLC. In: Official Methods and In: Crustacean Nutrition (D’Abramo, L.R., Conklin, D.E. &
Recommended Practices of the American Oil Chemists’ Society, Akiyama, D.M. eds), pp. 353–389. World Aquaculture Society,
5th edn, 2nd Printing (Firestone, D. ed.), official method Ce 1b- Baton Rouge, LA, USA.
89. AOCS Press, Champaign, IL, USA. Kanazawa, A. (2000) Nutrition and food. In: Spiny Lobsters: Fish-
Anderson, H.H. & Shubin, L.D. (1957) Improved Dumas method eries and Culture, 2nd edn (Phillips, B.F. & Kittaka, J. eds), pp.
for molecular weight determination. Anal. Chem., 29, 852–854. 611–624. Fishing News Books, Oxford, UK.
Association of Official Analytical Chemists (1995) Official Methods Kaplan, E.L. & Meier, P. (1958) Nonparametric estimation from
of Analysis. Association of Official Analytical Chemists, Wash- incomplete observation. J. Am. Stat. Assoc., 53, 457–481.
ington, DC, USA. Kittaka, J. (2005) Jellyfish as food organisms to culture phylloso-
Ates, R., Lindsay, D.J. & Sekiguchi, H. (2007) First record of an ma larva. Bull. Plankton Soc. Jpn., 52, 91–99 (in Japanese with
association between a phyllosoma larva and a Prayid siphono- English abstract).
phore. Plankton Benthos Res., 2, 67–69. Lucas, C.H. (1994) Biochemical composition of Aurelia aurita in
Booth, J.D., Webber, W.R., Sekiguchi, H. & Coutures, E. (2005) relation to age and sexual maturity. J. Exp. Mar. Biol. Ecol.,
Diverse larval recruitment strategies within the Scyllaridae. N. Z. 183, 179–192.
J. Mar. Freshwater Res., 39, 581–592. Mikami, S. & Kuballa, A.V. (2007) Factors important in larval
Bryan, G.W. (1971) The effect of heavy metals (other than Mer- and postlarval molting, growth, and rearing. In: The Biology
cury) on marine and estuarine organisms. Proc. R. Soc. Lond., and Fisheries of the Slipper Lobster, Crustacean Issue 17 (Laval-
B, Biol. Sci., 177, 398–410. li, K.L. & Spanier, E. eds), pp. 91–110. CRC Press, Boca Raton,
Canli, M. & Furness, R.W. (1995) Mercury and cadmium uptake FL, USA.
from seawater and from food by the Norway lobster Nephrops Murakami, K., Tamura, D., Awaji, M. & Takeuchi, T. (2011)
norvegicus. Environ. Toxicol. Chem., 14, 819–828. Nutritional value of blue mussel Mytilus galloprovincialis gonad
Chow, S., Suzuki, S., Matsunaga, T., Lavery, S., Jeffs, A. & for cultured larvae of Japanese spiny lobster Panulirus japonicus.
Takeyama, H. (2011) Investigation on natural diets of larval Aquacult. Sci., 59, 9–18 (in Japanese with English abstract).
marine animals using peptide nucleic acid-directed polymerase Nelson, M.M., Phleger, C.F., Mooney, B.D. & Nichols, P.D.
chain reaction clamping. Mar. Biotechnol., 13, 305–313. (2000) Lipids of gelatinous Antarctic zooplankton: Cnidaria and
Cox, S.L. & Johnston, D.J. (2003) Feeding biology of spiny lobster ctenophore. Lipids, 35, 551–559.
larvae and implications for culture. Rev. Fish. Sci., 11, 89–106. Nishida, S., Quigley, B.D., Booth, J.D., Nemoto, T. & Kittaka, J.
Cox, S.L., Bruce, M.P. & Ritar, A.J. (2011) Ingestion of artificial (1990) Comparative morphology of the mouthparts and foregut
diets with different textures as determined by the inert marker of the final-stage phyllosoma, puerulus, and postpuerulus of the
ytterbium oxide during culture of early-stage phyllosoma of the rock lobster Jasus edwardsii. J. Crust. Biol., 10, 293–305.
spiny lobster, Jasus edwardsii. Aquacult. Nutr., 17, 152–158. Ogushi, M. & Harada, R. (1997) Comparison between extractable
Davis, D.A. & Gatlin, D.M. III (1996) Dietary mineral require- components of salted and dried versus grilled and dried flying
ments of fish and marine crustaceans. Rev. Fish. Sci., 4, 75–99. fish. J. Jpn. Soc. Nutr. Food Sci., 50, 295–301 (in Japanese with
Fukuda, Y. & Naganuma, T. (2001) Potential dietary effects on English abstract).
the fatty acid composition of the common jellyfish Aurelia auri- Phillips, B.F. & Matsuda, H. (2011) A global review of spiny lob-
ta. Mar. Biol., 138, 1029–1035. ster aquaculture. In: Recent Advances and New Species in
Fukushi, K., Ishio, N., Tsujimoto, J., Yokota, K., Hamatake, T., Aquaculture (Fotedar, R.K. & Phillips, B.F. eds), pp. 22–84.
Sogabe, H., Toriya, K. & Ninomiya, T. (2004) Preliminary study Wiley-Blackwell, Oxford, UK.
on the potential usefulness of jellyfish as fertilizer. Bull. Soc. Sea- Phillips, B.F., Melville-Smith, R., Kay, M.C. & Vega-Velazquez,
water Sci. Jpn., 58, 209–217. A. (2013) Panulirus species. In: Lobsters: Biology, Management,
Goldstein, J.S. & Nelson, B. (2011) Application of a gelatinous Aquaculture and Fisheries, 2nd edn (Phillips, B.F. ed.), pp. 289–
zooplankton tank for the mass production of larval Caribbean 325. Wiley-Blackwell, West Sussex, UK.
spiny lobster, Panulirus argus. Aquat. Living Resour., 24, 45–51. Purcell, J.E., Baxter, E.J. & Fuentes, V.L. (2013) Jellyfish as prod-
Groeneveld, J.C., G~ oni, R. & Dıaz, D. (2013) Palinurus species. In: ucts and problems of aquaculture. In: Advances in Aquaculture
Lobsters: Biology, Management, Aquaculture and Fisheries, 2nd Hatchery Technology (Allan, G. & Burnell, G. eds), pp. 404–
edn (Phillips, B.F. ed.), pp. 326–356. Wiley-Blackwell, West Sus- 430. Woodhead Publishing, Cambridge, UK.
sex, UK. Saunders, M.I., Thompson, P.A., Jeffs, A.G., S€awstr€ om, C., Sach-
Herrnkind, W., Halusky, J. & Kanciruk, P. (1976) A further note likidis, N., Beckley, L.E. & Waite, A.M. (2012) Fussy feeders:
on phyllosoma larvae associated with medusae. Bull. Mar. Sci., phyllosoma larvae of the western rocklobster (Panulirus cygnus)
26, 110–112. demonstrate prey preference. PLoS ONE, 7, e36580(1–9).
Jeffs, A.G., Nichols, P.D. & Bruce, M.P. (2001) Lipid reserves Schoener, T.W. (1971) Theory of feeding strategies. Annu. Rev.
used by pueruli of the spiny lobster Jasus edwardsii in crossing Ecol. Syst., 2, 364–404.
..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
Sekiguchi, H., Booth, J.D. & Webber, W.R. (2007) Early life histo- Japanese Spiny Lobster (Ministry of Agriculture, Forestry and
ries of slipper lobsters. In: The Biology and Fisheries of the Slip- Fisheries of Japan eds), Kenkyuseika 480, pp. 38–45. Ministry of
per Lobster, Crustacean Issue 17 (Lavalli, K.L. & Spanier, E. Agriculture, Forestry and Fisheries of Japan, Tokyo, Japan (In
eds), pp. 69–90. CRC Press, Boca Raton, FL, USA. Japanese).
Simon, C.J., Carter, C.G. & Battaglene, S.C. (2012) Development Vijayakumaran, M. & Radhakrishnan, E.V. (2011) Slipper lob-
and function of the filter-press in spiny lobster, Sagmariasus sters. In: Recent Advances and New Species in Aquaculture
verreauxi, phyllosoma. Aquaculture, 370–371, 68–75. (Fotedar, R.K. & Phillips, B.F. eds), pp. 85–114. Wiley-Black-
Sims, H.W. Jr & Brown, C.L. Jr (1968) A giant scyllarid phylloso- well, Oxford, UK.
ma larva taken north of Bermuda (Palinuridea). Crustaceana, Wakabayashi, K. & Tanaka, Y. (2012) The jellyfish-rider: phyllo-
Suppl. 2 80–82. soma larvae of spiny and slipper lobsters associated with jelly-
Spanier, E. & Lavalli, K.L. (2013) Commercial scyllarids. In: Lobsters: fish. Taxa, Proc. Jpn. Soc. Syst. Zool., 33, 5–12 (in Japanese
Biology, Management, Aquaculture and Fisheries (Phillips, B.F. with English abstract).
ed.), 2nd edn, pp. 414–466. Wiley-Blackwell, West Sussex, UK. Wakabayashi, K., Sato, R., Hirai, A., Ishii, H., Akiba, T. &
Suzuki, N., Murakami, K., Takeyama, H. & Chow, S. (2006) Tanaka, Y. (2012a) Predation by the phyllosoma larva of Ibacus
Molecular attempt to identify prey organisms of lobster phyllo- novemdentatus on various kinds of venomous jellyfish. Biol. Bull.,
soma larvae. Fish. Sci., 72, 342–349. 222, 1–5.
Takahashi, M. & Saisho, T. (1978) The complete larval develop- Wakabayashi, K., Sato, R., Ishii, H., Akiba, T., Nogata, Y. &
ment of the scyllarid lobster, Ibacus ciliatus (von Siebold) and Tanaka, Y. (2012b) Culture of phyllosomas of Ibacus novemdent-
Ibacus novemdentatus Gibbes in the laboratory. Mem. Fac. Fish. atus (Decapoda: Scyllaridae) in a closed recirculating system
Kagoshima Univ., 27, 305–353 (in Japanese with English using jellyfish as food. Aquaculture, 330–333, 162–166.
abstract). Wang, M. & Jeffs, A.G. (2013) Nutritional composition of poten-
Takeuchi, T. (2010) Development of an effective artificial diet for tial zooplankton prey of spiny lobster larvae: a review. Rev.
phyllosoma. In: Development of Seed Production Technology in Aquacult., 5, 1–30.123

Appendix 1 Amino acid composition of the jellyfish species, Aurelia aurita and Chrysaora pacifica (mean  standard deviation; n = 3)
1
mg gDW % in total amino acids

Amino acid A. aurita C. pacifica A. aurita C. pacifica

Aspartic acid 3.54  0.16 b

6.10  0.39a
9.4  0.2a
8.6  0.2b
Threonine 1.90  0.08b 3.22  0.12a 5.0  0.2a 4.5  0.0b
Serine 1.76  0.11b 3.27  0.18a 4.6  0.1 4.6  0.1
Glutamic acid 5.21  0.21b 9.83  0.29a 13.8  0.3 13.9  0.1
Glycine 5.49  0.24b 11.76  0.48a 14.5  0.6b 16.6  0.3a
Alanine 2.54  0.16b 4.65  0.23a 6.7  0.3 6.6  0.1
Cystein 0.19  0.04b 0.31  0.02a 0.5  0.1 0.4  0.0
Valine 1.37  0.03b 2.57  0.03a 3.6  0.2 3.6  0.1
Methionine 0.57  0.07b 1.32  0.11a 1.5  0.1b 1.9  0.1a
Isoleucine 1.20  0.05b 2.31  0.04a 3.2  0.2 3.3  0.1
Leucine 1.69  0.80b 3.98  0.06a 4.4  2.0 5.6  0.1
Tyrosine 1.10  0.06b 2.15  0.05a 2.9  0.0b 3.0  0.1a
Phenylalanine 1.66  0.80 1.80  0.36 4.4  2.1 2.5  0.4
Lysine 2.58  0.13b 4.56  0.05a 6.8  0.1 6.4  0.3
Histidine 0.44  0.03b 0.97  0.03a 1.2  0.0b 1.4  0.1a
Arginine 2.62  0.09b 4.54  0.30a 6.9  0.2a 6.4  0.2a
Proline 3.93  0.15b 7.56  0.18a 10.4  0.3 10.7  0.2
Essential amino acids 14.02  1.20b 25.27  0.95a 36.6  1.6 34.8  0.3
Total 37.77  2.00b 70.90  2.72a 100 100

Different superscript letters on each line indicate a significant difference (P < 0.05).

..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
 Appendix 2 Fatty acid composition of the jellyfish species, Aurelia aurita and Chrysaora pacifica (mean  standard deviation; n = 3)
1
mg gDW % in total fatty acids

Fatty acid A. aurita C. pacifica A. aurita C. pacifica

C11:0 0.002  0.001 0.004  0.001 <0.1  0.0 <0.1  0.0

C12:0 0.019  0.009 0.031  0.015 0.4  0.1 0.4  0.1
C14:0 0.122  0.029b 0.262  0.077a 2.8  0.1b 3.7  0.1a
C15:0 0.089  0.023b 0.191  0.042a 2.1  0.3b 2.7  0.2a
C16:0 0.947  0.144 1.043  0.158 22.5  2.2a 14.9  1.8b
C16:1 0.190  0.026b 0.414  0.064a 4.5  0.7 5.9  1.0
C17:0 0.225  0.031 0.323  0.104 5.4  1.0 4.5  0.8
C17:1 0.033  0.015 0.040  0.014 0.8  0.2 0.6  0.1
C18:0 0.817  0.162b 1.247  0.161a 19.2  0.2 18.1  4.2
C18:1 0.262  0.056 0.381  0.253 6.2  0.3 5.1  3.2
C18:2 0.056  0.018 0.056  0.055 1.3  0.2 0.7  0.5
C18:3 0.038  0.019 0.059  0.028 0.9  0.3 0.8  0.4
C18:4 0.081  0.012 0.118  0.026 1.9  0.4 1.7  0.3
C19:0 0.029  0.004 0.074  0.031 0.7  0.2 1.0  0.2
C20:0 0.012  0.001 0.043  0.032 0.3  0.0 0.6  0.3
C20:1 0.026  0.002 0.080  0.078 0.6  0.2 1.0  0.8
C20:3 0.004  0.002 0.010  0.010 <0.1  0.0 0.1  0.1
C20:4 0.336  0.086 0.983  0.373a 7.8  0.4b 13.5  1.7a
C20:5 0.760  0.273 1.104  0.429 17.5  3.0 15.2  2.1
C22:1 0.004  0.001 0.028  0.009a 0.1  0.0b 0.4  0.0a
C22:6 0.055  0.015 0.232  0.080a 1.3  0.2b 3.2  0.3a
Unidentified 0.156  0.037 0.419  0.141a 3.6  0.4b 5.8  1.3a
Essential fatty acids 1.245  0.403 2.435  0.925 28.7  3.8 33.5  3.5
Polyunsaturated fatty acids 1.330  0.411 2.562  0.955 30.7  3.6 35.3  3.4
Monounsaturated fatty acids 0.516  0.094 0.944  0.363 12.2  1.0 13.0  3.0
Saturated fatty acids 2.261  0.371 3.220  0.574 53.4  3.3 45.9  5.9
Total 4.263  0.882 7.145  1.935 100 100

Different superscript letters on each line indicate a significant difference (P < 0.05).

Appendix 3 Mineral composition of the jellyfish species, Aurelia aurita and Chrysaora pacifica (mean  standard deviation; n = 3)

mg gDW 1
or lg gDW 1
% in total minerals

Mineral A. aurita C. pacifica A. aurita C. pacifica

Major minerals (mg gDW 1)

Ca 4.583  0.173a 4.164  0.061b 1.076  0.058 1.154  0.075
Na 373.109  18.987a 317.661  18.580b 87.452  0.241 87.828  0.714
Mg 32.459  1.323a 27.284  0.040b 7.612  0.177 7.559  0.409
K 16.457  0.709a 12.480  0.267b 3.861  0.162 3.459  0.231
Total 426.608  20.588a 361.589  18.325b 100 100
Trace minerals (lg gDW 1)
B 7.759  2.995 8.475  3.739
Al 0.312  0.161 0.279  0.142
V 0.016  0.004 0.024  0.009
Cr 0.186  0.171 0.357  0.322
Mn 0.159  0.037 0.116  0.049
Fe 0.599  0.313 1.427  0.605
Cu 0.136  0.049 0.210  0.035
Zn 4.004  0.947 3.644  4.062
As 1.371  0.133 1.413  0.076
Se 5.490  0.565 5.251  0.242
Mo 0.021  0.007 0.035  0.020
Cd 0.002  0.001 0.006  0.004
Hg 0.023  0.011 0.020  0.007
Pb 0.009  0.002 0.007  0.004

Different superscript letters on each line indicate a significant difference (P < 0.05).

..............................................................................................

Aquaculture Nutrition, 22; 25–33 ª 2015 John Wiley & Sons Ltd
Copyright of Aquaculture Nutrition is the property of Wiley-Blackwell and its content may
not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's
express written permission. However, users may print, download, or email articles for
individual use.