Aquaculture Research, 2007, 38, 1082^1087 doi:10.1111/j.1365-2109.2007.01775.

Growth of the hatchery-produced juveniles of

commercial sea cucumber Holothuria (Theelothuria)
spinifera Theel

P S Asha & P Muthiah

Tuticorin Research Centre of Central Marine Fisheries Research Institute,Tuticorin,Tamil Nadu, India

Correspondence: P S Asha, Tuticorin Research Centre of Central Marine Fisheries Research Institute, South Beach Road Extension,
Tuticorin 628 001,Tamil Nadu, India. E-mail: [email protected]

Abstract stocks in many countries have resulted in severe

overexploitation of commercially important species.
The present study was conducted to develop the
Bruckner, Johnson and Field (2003) reported that the
hatchery technology for seed production of the
holothurians are susceptible to overexploitation be-
widely exploited commercial sea cucumber Holothur-
cause of late maturity, density-dependent reproduc-
ia spinifera Theel 1886, to facilitate an e¡ective stock
tion and low rate of recruitment. The release of
enhancement programme. Broodstock collected by
hatchery-produced juvenile sea cucumbers into the
skin divers were used for spawning trials. The
natural habitat is being advocated in many countries
embryonic and larval stages were similar to other
as one of the means of rebuilding the wild stock, a
aspidochirotes. The larval survival rate was
process termed as restoration, restocking or reseed-
43.5% on day 9, which decreased to 18.3% nearing
ing (Battaglene 1999).
metamorphosis on day 13, the growth rate was
The breeding and cultivation of holothurians dates
48 mm day
1 and the settlement rate was 3.5%. Alga-
back to 1950, with the ¢rst recorded production of
macs used to induce settlement also served as the
juveniles of Stichopus japonicus (Imai, Inaba, Sato &
food source for the early settled juveniles, followed
Hatanaka 1950). Later, studies on reproduction, early
by Sargassum spp. extract (o40 mm) for the ¢rst
development and metamorphosis of Cucumaria elon-
month. Sargassum spp. powder along with ¢ne sand
gata (Chia & Buchanan 1969), C. pseudocurata
(1:1) was given to juveniles 420 mm. Addition of Spir-
(Rutherford 1973), Aslia lefevrai (Costelloe 1988), Pso-
ulina spp. along with Sargassum spp. powder and ¢ne
lus chitonoides and P. bullatum (McEuen & Chia 1991)
sand (0.5:1:2) enhanced the growth rate of the juve-
and C. frondsa (Hamel & Mercier 1996) were exten-
niles. In the hatchery, the juveniles attained an aver-
sively undertaken. Meanwhile in 1985, techniques
age size of 1, 30 and 48 on 20, 80 and 120 days
were developed for producing juveniles of S. japonicus
respectively.
in China (Shuxu & Gongchao 1981; Chen 2003) and
seed of Actinopyga echinites in Taiwan (Chen & Chian
Keywords: Holothuria spinifera, survival rate, Al-
1990).
gamacs, Sargassum spp., Spirulina spp.
Following the Chinese method, James, Rajapan-
dian, Baskar and Gopinathan (1988) successfully pro-
duced the seed of Holothuria scabra for the ¢rst time.
Introduction
This technique was later applied for the seed produc-
Sea cucumbers belonging to the families Holothuri- tion of sea cucumbers in Australia, Indonesia, Mald-
dae and Stichopodidae form an important part of ives and Solomon Islands (Battaglene 1999). The
multispecies ¢shery, existing for over 1000 years seeds of Isostichopus £uscus and H. fuscogliva have
along the Indo-Paci¢c region, and the processed pro- been successfully produced in Ecuador and in Kirbati
duct is a valuable source of income. Increasing de- Island (Hamel, Hidalgo & Mercier 2003; Friedman &
mand and inadequate management of sea cucumber Tekanene 2005).

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1082 Journal Compilation r 2007 Blackwell Publishing Ltd
Aquaculture Research, 2007, 38, 1082^1087 Hatchery-produced juveniles of commercial sea cucumber P S Asha & P Muthiah

In India, the beche de-mer industry mainly de- For holothurian seed production150 000 fertilized
pends on two species, H. scabra and H. spinifera (Chel- eggs, 5700 pentactulae and 2000 juveniles were
laram, Samuel & Patterson Edward 2003), and both reared in a lot following the hatchery technique
the species are being processed for export. Holothuria (James et al.1988; Battaglene1999; Chen 2003).While
spinifera, locally called cheena attai or raja attai, was rearing, periodically representative samples were
once rated very high in the market and was in good measured for assessing the growth of larvae and ju-
demand in China. At present, the market value is veniles. The microalgae cultured by the serial dilu-
moderate; the processed ones (60 counts kg
1) fetch tion technique (Gopinathan 1982) were provided as
US $37.5. Besides Gulf of Mannar and Palk Bay of In- larval feed as indicated in Asha and Muthiah (2006)
dian coast, this species is reported to occur in the Red and at a concentration speci¢ed byAsha (2004).
sea, Persian Gulf, Sri Lanka, North Australia, Philip- The doliolaria larvae on 10th day were maintained
pines and China (James 1995). James and Badrudeen in a £owthrough system, in which an equal volume of
(1997) reported an estimated annual landing of 460 seawater released was let in and were provided with
tonnes of fresh H. spinifera along the south-east coast Algamacs (Bio-Marine, Hawthrone, CA, USA) at a
of India. Increase in demand and inadequate ¢shery rate of 0.5 g 500 L
1 for inducing them to settle as
management has resulted in overexploitation of com- pentactulae. The Algamacs was provided for13 days
mercial sea cucumber species in India also; hence, until the juveniles became visible to the naked eye.
the Ministry of Environment, Government of India, For the newly settled juveniles, Sargassum spp. ex-
has banned both ¢shery and export of sea cucumbers tracts (o40 mm) were given (10 L 500 L
1) for 4
since 2001, which has had a severe impact on the li- weeks.When the juveniles attained an average length
velihood of poor coastal communities subsisting on of 20 mm, a mixture of Sargassum spp. powder and
its ¢shery and processing. Owing to the role played ¢ne sand in a proportion of (1:2) was given at 1% of
by H. spinifera in the commercial ¢shery and also the body weight of juveniles (initially o80;
due to its indiscriminate exploitation, knowledge of o200 mm as the days progressed). From day 30, Spir-
early development, larval and juvenile rearing is es- ulina spp. (15 mg juvenile
1) was provided along
sential for the seed production so as to carry out, the with the above feed. Every week, juveniles were taken
captive or wild stock enhancement programme of out using a ¢ne brush and the length and number of
this species e¡ectively. Hence, an attempt was made live juveniles were noted for assessing the growth
to produce juveniles of H. spinifera in the laboratory. and survival rate. Once in a week, the juveniles were
The present work is the ¢rst successful attempt of ju- transferred to another tank as suggested by Batta-
venile production of this species under laboratory glene (1999). The juveniles thus produced from ferti-
conditions. lized eggs were reared up to 120 days.

Materials and methods

Results and discussion
The broodstock of H. spinifera collected by skin divers
Spawning and development
were maintained in 1000 L FRP tanks having
150 mm thickness of ¢ne coral beach sand at the bot- The induced spawning on 2 January 2003 occurred
tom. The water was exchanged daily and sand every in the afternoon, after addition of feed. There was no
fortnight and the brooders were fed with Sargassum spawning by thermal stimulation. The female
spp. powder at the rate of 5% of their body weight. spawned ¢rst, followed by three males.
For induced spawning, among the 30 numbers of The spawning in H. spinifera was induced by addi-
broodstock, 10 numbers were subjected to thermal tion of feed. Addition of Algamacs at 50 g 500 L
1
stimulation wherein the holothurians were exposed induced spawning in H. fuscogilva (Battaglene, Sey-
to 3^4 1C above the ambient temperature for an hour. mour & Ramofa¢a 2002). Thermal stress, despite
The high temperature was maintained by a heating having been e¡ective for S. japonicus (Ishida 1979),
element controlled by a thermostat. Similarly, indu- H. scabra (James et al.1988) and H. scabra and A. maur-
cement by addition of feed constituted by rice bran, itiana (Battaglene et al. 2002), did not induce H. spini-
Soya powder and Sargassum spp. powder (4:1:2) at a fera to spawn. Similarly, Battaglene et al. (2002)
rate of 50 g 500 L
l was also attempted to 10 num- observed that thermal stress was not e¡ective but ad-
bers and the remaining10 numbers were maintained dition of dried algae alone yielded induced spawning
as control. in H. fuscogilva.

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Journal Compilation r 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1082^1087 1083
Hatchery-produced juveniles of commercial sea cucumber P S Asha & P Muthiah Aquaculture Research, 2007, 38, 1082^1087

In the successful spawning experiment, the fe- by 20% for periphytic diatom-fed larvae. Ito and
males spawned ¢rst, followed by the males. Because Kitamura (1997) accelerated the metamorphosis in
the released gonadal materials instigate the mass S. japonicus by providing periphytic diatom at
spawning, the blended gonad from mature brood- 420 000 cells cm
2. Studies have to be undertaken
stock may be an e¡ective spawning stimulant (Batta- to evolve suitable cues and their concentration so as
glene 1999). to obtain more juveniles.
The spawning behaviour of H. spinifera and the On day 15, 5700 pentactulae of 0.32 mm length,
occurrence of spawning in the afternoon were simi- forming 9.4% of early auricularia, were produced.
lar to the observations on other aspidochirotids On day 23, 2000 juveniles of 0.95 mm, forming 3.5%
(McEuen 1988). The spawning in the late afternoon of early auricularia, were reared. On day 31, the juve-
and evening may be related to a decrease in light in- niles had a mean length of 2.3 mm (Fig. 2a) and the
tensity as opined by McEuen (1988). Fertilization and survival rate was 7.5%. On day 48, the length of
early embryonic development and other larval stages the juveniles ranged from 6 to 14 mm, with a mean
of H. spinifera are similar to those of other aspidochir- of 10.1mm (Fig. 2b) and the survival rate was 3.5%
ote holothurians (Ishida1979; Maruyama1980; Chen (Fig. 3). Adding Spirulina spp. along with Sargassum
& Chian 1990). spp., a growth rate of 1.5 mm day
1 was obtained.
On day 85, 56 juveniles produced in the size range of
31^60 mm with a mean of 41mm (Fig. 2c) were pro-
Larval and juvenile rearing
duced. The observed growth rate was 0.64 mm day
1
The chronological developmental stages and the de- with a 2.8% survival. In the fourth month, the juve-
tails of the various larval stages up to pentactulae niles attained a mean size and weight of 48 mm and
with the time of occurrence and the mean size were 2.84 g respectively (Fig. 2d).
as described byAsha and Muthiah (2002). The growth of the juveniles can be described by
The doliolaria stage was attained on day 10 in H. the following equation:
spinifera, which was similar in H. scabra (James et al.
Y ¼ 0:0001x2:8513
1988; Battaglene, Seymour & Ramofa¢a1999). It took
15 days at 25^28 1C to reach doliolaria in A. echinites An r 5 0.993 indicated a high level (Po0.01) of sig-
(Chen & Chian 1990). During the rearing period of ni¢cance (Fig. 4).
the larvae and juveniles, the seawater temperature During the juvenile rearing, maximum mortality
ranged from 29 to 31 1C, salinity from 34.8 to 36 ppt, (92.5%) occurred on day 31 at 2.3 mm length. Batta-
pH from 8.1 to 8.2 and dissolved oxygen from 4.1 to glene et al. (1999) observed that juveniles of o5 mm
5.2 mL
1. The attainment of doliolaria on day 10 for holothurians are more susceptible. The juveniles at-
H. spinifera may be due to the larval rearing at a high- tained an average size of 41mm (2.3 gm) on day
er temperature of 29^31 1C. Asha and Muthiah 85, with an average growth rate of 0.64 mm day
1.
(2005) indicated, a water temperature of 28^32 1C as
the optimum for rearing the larvae of H. spinifera.
During the spawning trial, 43.8% of auricularia 1100
1050
were hatched out from 1500 000 fertilized eggs. On
1000
day 3, the survival rate was 79.9%, which decreased
950
to 63.9% and 43.5% on days 5 and 9 respectively. On 900
day 11, the survival was 39.6%. Nearing metamor- 850
Length (µm)

phosis, on day 13, the survival rate was 18.3%. 800

The larval growth rate was 48 mm day
1 and the 750
growth equation indicated a high level of signi¢- 700
650
cance (r 5 0.9695; Po0.01) (Fig. 1).
600
The metamorphosis and settlement are the critical y = 379.78x0.4211
550 2
stages in the development of sea cucumber larvae 500 R = 0.9399
(Preston 1993). In this study, Algamacs was used as 450
the settlement cue. Battaglene (1999) also observed 400
0 2 4 6 8 10 12 14
high settlement using Algamacs in H. scabra. In
Days
H. spinifera, Asha and Muthiah (2002) reported more
settlement (40%) of Algamacs-fed larvae, followed Figure 1 Growth rate of the larvae of Holothuria spinifera.

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1084 Journal Compilation r 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1082^1087
Aquaculture Research, 2007, 38, 1082^1087 Hatchery-produced juveniles of commercial sea cucumber P S Asha & P Muthiah

Figure 2 Juveniles of Holothuria spinifera (a) 31 days old, (b) 48 days old, (c) 85 days old, (d) 120 days old.

100 50

90 45

80 40

70 35
Length (mm)
Survival (%)

60 30
25
50
20
40
15 2.8513
30 y = 0.0001x
10 2
20 R = 0.9864
5
10
0
0 0 20 40 60 80 100
23 28 31 38 42 48 55 64 75 80 87 Days
Days
Figure 4 Growth rate of the juveniles of Holothuria
Figure 3 Survival rate of the juveniles of Holothuria spi- spinifera.
nifera.

Battaglene et al. (1999) recorded a 0.5 mm growth duced in this study, after 120 days of growth in the
rate for H. scabra. The growth rate of H. spinifera was hatchery, were sea ranched into the sea grass beds of
10^20 mm greater in 3 months and was similar to I. Van Island area in Gulf of Mannar at 10 m depth for
japonicus (Ito & Kitamura 1998). The average size of the ¢rst time. In this study, addition of Spirulina spp.
the juveniles sea ranched reported was 20^30 mm increased the growth rate to 1.4 mm day
1 and it in-
by James, Gandhi, Palaniswamy and Rodrigo (1994); dicated that the probable role of additional protein
o60 mm and 20 g by Battaglene (1999); and 35 mm source in the growth of juveniles and powdered algae
by Hamel et al. (2003). The juveniles of H. spinifera is not a major source of food for juveniles (Battaglene
(mean length and weight 48 mm and 2.84 g) pro- et al. 1999). The feeding habit of the juvenile sea

r 2007 The Authors

Journal Compilation r 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1082^1087 1085
Hatchery-produced juveniles of commercial sea cucumber P S Asha & P Muthiah Aquaculture Research, 2007, 38, 1082^1087

cucumber in the wild is unknown (Battaglene et al. Chen C.P. & Chian C.S. (1990) Short note on the larval devel-
1999). Moreover, Yingst (1978) opined that many de- opment of the sea cucumberActinopyga echinites (Echino-
posit-feeding holothurians do not appear to assimi- dermata: Holothuroidea). Bulletin of Institute of Zoological
late algae and have little cellulose activity in their Academic Sinica 29,127^133.
gut. Hence, formulation of a suitable balanced diet Chia F.S. & Buchanan J.B. (1969) Larval development of Cu-
cumaria elongata (Echinodermata: Holothuridea). Journal
for juveniles of sea cucumbers has to be given more
of Marine Biological Association of United Kingdom 49,
thrust in future studies.
151^159.
Costelloe J. (1988) Reproductive cycle, development and re-
cruitment of two geographically separated populations
Acknowledgments of the dendrochirote holothurian Aslia lefevrei. Marine
Biology 99, 535^545.
The authors express their sincere gratitude to Prof. Friedman K. & Tekanene M. (2005) White teat¢sh at
(Dr) Mohan Joseph Modayil (Director) and Dr M. Ra- Kiribati sea cucumber hatchery ‘‘Local technicians getting
jagopalan (Head of Fishery Environment and Man- them out again’’. SPC Beche-de-mer Information Bulletin 21,
agement Division) of CMFRI, Kochi, for their interest 32^33.
and encouragement. Gopinathan C.P. (1982) Methods of culturing phyto-
plankton. In: Manual of Research Method for Fish and
Shell¢sh Nutrition. Central Marine Fisheries Research
Institute, Kochi, India. Special Publication 8, 113–118.
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