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Received: 13 January 2021    Revised: 14 March 2021    Accepted: 16 April 2021

DOI: 10.1111/anu.13274

ORIGINAL ARTICLE

The effects of dietary supplement of Ulva fasciata, fermented

soybean and probiotics on the growth of Apostichopus
japonicus

Zhen-­Hao Liao1 | María Joaquina Ibarra-­Arana1 | Jian-­Cheng Chen1 | Huai-­Ting Huang1 |

Yu-­Ju Lin2 | Fan-­Hua Nan1

1
Department of Aquaculture, National
Taiwan Ocean University, Keelung, Taiwan
2
Department of Life Sciences, National
Chung Hsing University, Taichung, Taiwan
Correspondence
Fan-­Hua Nan, Department of Aquaculture,
National Taiwan Ocean University, No.
2, Beining Road, Jhong-­jheng District,
Keelung 20224, Taiwan, ROC.
Email: fhnan@mail.ntou.edu.tw
Abstract
Several feeding experiments were conducted to evaluate the effects of dietary sup-
plement of seaweed, fermented soybean and probiotics on the growth of juvenile
sea cucumber Apostichopus japonicus. After comparing the growth performance of
sea cucumbers fed with Ulva fasciata, Gracilaria tenuistipitata and Laminaria japonica
for 8 weeks, U. fasciata was selected as a major protein source. Four partial replace-
ments of U. fasciata in feeds were then prepared by the inclusion of white fish meal,
soybean meal, mixture of fish and soybean meal and fermented soybean (BioMax®),
respectively. The growth performance over a period of 8 weeks revealed that sea
cucumbers fed with fermented soybean supplemental diet had significantly better
weight gain (129.60 ± 0.14%), SGR (1.48 ± 0.01%) and FCR (2.93 ± 0.01) compared
with individuals fed with other diets (p < .05). Moreover, the relative higher attractive-
ness of fermented soybean supplemental diet was shown in food choice experiments
(pairwise comparison). Daily addition of 109 CFU of probiotics mixture (Bacillus subtilis
and Micrococcus luteus) in diet for 8 weeks could be suitable for the growth of sea
cucumbers while sand was used as a substrate in tank bottom. The present study pro-
vided useful information for tank-­based aquaculture of Apostichopus japonicus.

KEYWORDS
Apostichopus japonicus, attractiveness, fermented soybean, growth, probiotics, Ulva fasciata

1  |  I NTRO D U C TI O N Yu et al., 2015). In previous studies, seaweed Sargassum thunbergii

Among echinoderms, sea cucumbers are commercially important re-
source because of their medicinal and nutritious benefits (Bordbar
et al., 2011; Pangestuti & Arifin, 2018). There are approximately
1250 identified species in Asia-­Pacific (Du et al., 2012). Since the
market demand for Apostichopus japonicus has increased and the
wild production of this species has declined due to over-­exploitation,
the improvement of culture conditions and health management ap-
proach have been developed in Korea (Seo & Lee, 2011; Seo et al.,
2011a, 2011b) and China (Sun et al., 2018; Wen et al., 2016; Xia, Gao,
et al., 2015; Xia, Wang, et al., 2015; Xia et al., 2018; Yang et al., 2016;
was commonly used as a major ingredient for feeding A.  japonicus
(Liu et al., 2009; Yuan et al., 2006). However, this algal species is not
widely distributed and produced commercially in subtropical area.
Therefore, finding alternative could be helpful for the sustainability
of sea cucumber farming in Taiwan.
In comparison with traditional sea cucumber feed S. thunbergii,
various alternatives were suggested with the characteristic of lower
cost, higher availability and be more preferable to the nutritional re-
quirements of sea cucumber (Gao et al., 2011; Jia et al., 2019; Seo
et al., 2011a, 2011b; Wen et al., 2018). It has been reported that
growth performance of sea cucumber can be much improved by

Aquaculture Nutrition. 2021;00:1–7. wileyonlinelibrary.com/journal/anu© 2021 John Wiley & Sons Ltd     1 |
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2      LIAO et al.
dietary supplement of fermented soybean (FSB) (Seo et al., 2011b),
solid-­state fermented diet (Wang, Guo, et al., 2017) and hydrolysed
soybean meal (Song et al., 2016). Moreover, several studies demon-
strated that dietary probiotics had positive effects on the growth
and immunity of sea cucumber (Chi et al., 2014; Ma et al., 2018;
Yang et al., 2015). In the present study, serial feeding trials were con-
ducted to determine the proper dietary supplement for Apostichopus
japonicus (Selenka, 1867). The feeding preference and growth of sea
cucumber were studied by using seaweed, commercial FSB and pro-
biotics mixture as feed ingredients.
2  |  M ATE R I A L S A N D M E TH O DS
2.1  |  Sea cucumber rearing
The experiments were carried out at the Department of Aquaculture,
College of Life Science, National Taiwan Ocean University (NTOU).
Apostichopus japonicus were obtained from the Aquatic Animal Center
(AAC-­NTOU) and maintained in glass tanks (60 cm × 40 cm × 40 cm,
96 L) with a flow-­through water system. Sea cucumbers were fed
once daily at 17:00 h ad libitum. Seawater quality parameters were
maintained at temperature 22 ± 1°C and salinity 33 ± 1 g/L.
2.2  |  Materials
Fresh Ulva fasciata and Gracilaria tenuistipitata were cultured in FRP
barrels of 1-­ton water capacity at AAC-­NTOU. The algae were washed
and then dried in the oven at 60°C for 12 h. After that, the dried algae
were pulverized and sieved (40 µm mesh). Laminaria japonica powder
was purchased from Chuan Kuan Enterprise Co., Ltd. The algal pow-
der was stored at 4°C until use. Proximate composition analysis of
all algal powder was conducted according to AOAC (1995) methods
(Table 1). White fish meal (645 g/kg crude protein and 125 g/kg crude
lipid), soybean meal (470 g/kg crude protein and 33 g/kg crude lipid),
gelatinized cellulose and Maltodextrin were purchased from Taisun
®
Co., Ltd. Fermented soybean meal BioMax (255 g/kg crude protein
and 29 g/kg crude lipid on dry matter basis, 420 ± 20 g/kg moisture,
30–­40 g/kg fibre, <60 g/kg ash, Lactobacillus sp. (>5 × 109 CFU/ml)
and acidic protease (>20,000 U/kg)) was purchased from Grobest
International Co., Ltd.. Substrate was prepared using the mixture
of coral and shell sand for the probiotic feeding trial. The sand was
sieved to homogenized size of particles (≤0.84 mm) and then spread
in the bottom of experimental tank (5 cm thickness).
2.3  |  Experimental diets preparation
The three algal powders (U. fasciata, G. tenuistipitata and L. japonica)
were referred as U, G and L, respectively. On the basal level of 40
percent Ulva inclusion, the experimental diets with different supple-
ment were prepared including, F (150 g/kg White fish meal inclusion),
TA B L E 1  Proximate composition of Ulva fasciata, Gracilaria
tenuistipitata and Laminaria japonica powder
Macroalgae
Proximate Ulva Gracilaria Laminaria
composition (g/kg) fasciata tenuistipitata japonica
Moisture 71.8 64.4 80.2
Crude protein 157.0 171.0 102.3
Crude lipid 22.6 14.0 12.5
Crude fibre 114.8 78.0 91.1
Ash 224.4 154.0 289.8
S (200 g/kg Soybean meal inclusion), M (75 g/kg White fish meal and
103  g/kg Soybean meal inclusion) and B (380  g/kg BioMax® inclu-
sion). All ingredients with given proportions were watered, stirred
and extruded using a pelletizer. The pellets were dried at 40°C for
12 h. Once it cooled, the feed was grinded into smaller pieces and
stored at 4°C until use. Ingredients and nutrient contents were an-
alysed according to AOAC (1995) methods (Table 2). Gross energy
(kcal/kg diet) was measured by Parr 6200 calorimeter. Bacillus sub-
tilis and Micrococcus luteus which obtained from Dr. Huang Shi-­Ling
(Freshwater Aquaculture Research Center of the Fisheries Research
Institute of Taiwan, Council of Agriculture [COA]) were used in the
probiotics supplement. B. subtilis and M. luteus were previously iso-
lated from the freshwater shrimp pond. Genetic characterizations
were achieved by 16S rDNA sequence analysis. Their beneficial ef-
fects on the improvement of water qualities as well as shrimp and
fish growth have been reported in Chinese journals and theses. Both
strains were cultured in tryptic soy broth (TSB) at 30°C. The probiotic
was prepared daily at 1:1 mixture of B. subtilis (6.248 × 108 CFU/ml,
OD600 = 0.9) and M. luteus (5.437 × 108 CFU/ml, OD600 = 1) and then
added to experimental diet forming a final dosage of 1 × 109 CFU/g.
2.4  |  Feeding trial
The feeding trial was organized in three sequential experiment with
recirculating system. In first experiment, three algal powders (U, G
and L) were tested to determine the optimal food for sea cucumber.
Ninety sea cucumbers (initial weight of 60 g) were randomly allocated
to nine FRP barrels (45 cm × 0.63 m2) in equal numbers (n = 10) to
yield three groups in triplicate. In second experiment, sea cucumbers
were fed with five types of diets with the ingredient of either opti-
mal algal powder from first experiment or mixture of optimal algal
powder and various protein sources described in Section 2.3 (F, S, M
and B). One hundred fifty sea cucumbers (initial weight of 10 g) were
randomly allocated to 15 tanks (45  cm  ×  30  cm  ×  30  cm) in equal
numbers (n = 10) to yield five groups in triplicate. In third experiment,
the optimal diet from second experiment with or without the addition
of probiotics was used for feeding sea cucumbers in bare tank or tank
with a sand substrate. Seventy-­t wo sea cucumbers (initial weight of
30 g) were randomly allocated to 12 tanks (45 cm × 40 cm × 28 cm)
in equal numbers (n = 6) to yield four groups in triplicate. All feeding
LIAO et al. |
      3

TA B L E 2  Ingredient (dry matter basis)

Diets
and proximate composition of Ulva-­based
experimental diets U F S M B
†
Ingredients (g/kg)
U. fasciata powder 1000 400 400 400 400
White fish meal -­ 150 -­ 75 -­
Soybean meal -­ -­ 200 103 -­
BioMax -­ -­ -­ -­ 380
Cellulose -­ 250 200 222 20
Dextrin -­ 200 200 200 200
Proximal composition (g/kg)
Moisture 71.8 74.2 76.5 78.5 76.1
Crude protein 157.0 172.1 169.1 166.7 159.7
Crude lipid 22.6 17.9 15.0 16.2 17.3
Crude fibre 114.8 217.8 206.4 217.1 131.5
Ash 224.4 119.4 104.0 112.7 111.2
‡
NFE 409.4 398.6 429.0 408.8 504.2
Gross energy 2927.6 3526.7 3493.7 3699.8 3392.8
(kcal/kg diet)
†
The protein contents of ingredients (dry matter basis): U. fasciata powder (157.0 g/kg), White
fish meal (644.9 g/kg), Soybean meal (470.5 g/kg) and BioMax (255.0 g/kg); The lipid contents
of ingredients (dry matter basis): U. fasciata powder (22.6 g/kg), White fish meal (125.3 g/kg),
Soybean meal (32.5 g/kg) and BioMax (29.1 g/kg).
‡
NFE (Nitrogen-­free extract): [1000 − (Moisture + Crude protein + Crude lipid + Crude fibre + Ash)]

trials were conducted for 8 weeks. During the experimental period,
the seawater temperature was maintained at 22 ± 1°C, salinity ranged
between 32–­33 g/L, ammonia nitrogen level below 0.25 mg/L and
dissolved oxygen above 5.0 mg/L. Sea cucumbers were fed five per
cent of wet body weight once a day at 17:00 h. At the end of 8 weeks,
the following parameters were measured:
[
(Final weight − Initial weight)
] Test number
Weight gain ( % ) = × 100
Initial weight
[ ( ) ( )]
ln Final weight (g) − ln Initial weight (g)
Specific growth rate (SGR) ( % ) =
day (s)
selected sea cucumbers were placed in the centre of experimental
tank and the attractive number in each diet was recorded after 18 h.
All measurements (10 comparison tank) were performed at the same
time. Second and third repeat were conducted sequentially at 2-­day
intervals. The attractiveness was calculated as following:
[ ]
Attractive number
Relative attractiveness ( % ) = × 100
(1)
Average attractiveness ( % ) = The average of relative attractiveness
× 100
2.6  |  Statistical analysis
(2)

Each experiment was repeated in triplicate, and data were shown as

feed intake (g)
Feed conversion ratio (FCR) = ( mean ± SD (standard deviation). Statistical analysis was performed
final weight (g) − initial weight (g) (3)
)
using SAS (Statistical Analysis System) Software version 9.0. p < .05
( ) was considered as significant difference as the result of the one-­way
Final number of sea cucumber ANOVA with a subsequent Least Significant Difference (LSD) test.
Survival ( % ) = × 100 (4)
Initial number of sea cucumber

2.5  |  Relative attractiveness 3  |  R E S U LT S

The relative attractiveness among five diets (U, S, F, M and B) was
determined by pairwise comparison (Ibarra-­Arana et al., 2018). Each
comparison was conducted in one tank (150 cm × 50 cm × 30 cm) with
two different diets (5 g) placing in opposite corners. Seventy experi-
mental animals with an average wet weight of 20 ± 5 g were reared in
closed seawater at 22 ± 1°C. After starvation for 24 h, seven randomly
3.1  |  Effects of seaweed-­based diets on the growth
performance of Apostichopus japonicus
Growth performance of sea cucumber fed three seaweed power
(U.  fasciata [U], G.  tenuistipitata [G], L.  japonica [L]) was shown in
Table 3. The survival rates were 100% in each dietary group at the
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4      LIAO et al.

TA B L E 3  Effects of dietary Ulva fasciata (U), Gracilaria tenuistipitata (G), and Laminaria japonica (L) on the growth performance of sea
cucumber (Apostichopus japonicus) for 8 weeks

Survival
Diet Initial weight (g) Final weight (g) Weight gain (%) SGR (%) FCR FI (g) rate (%)

U 62.38 ± 0.54a 84.38 ± 7.88a 35.20 ± 11.51a 0.53 ± 0.15a 5.35 ± 1.89a 108.49 ± 3.97a 100

a b b b b
G 62.16 ± 3.40 70.96 ± 1.16 14.34 ± 5.31 0.24 ± 0.08 12.74 ± 3.68 105.62 ± 3.92a 100
a b b b b b
L 61.63 ± 1.11 67.81 ± 2.22 10.02 ± 1.96 0.17 ± 0.03 16.62 ± 3.40 99.86 ± 1.67 100

Note: (1) Values are presented as mean ± SD. (2) Means in the same column with the different superscripts (a, b) are significantly different (p < .05).
(3) Weight gain: [(Final weight − Initial weight)/Initial weight] × 100%. (4) SGR (specific growth rate): [ln (Final weight (g)) − ln (Initial weight (g))/
day (s)] × 100%. (5) FCR (feed conversion ratio): feed intake (g)/(final weight (g) − initial weight (g)). (6) FI (feed intake): feed supplied (g dry
weight). (7) Survival rate: (Final number of sea cucumber/Initial number of sea cucumber) × 100%.

TA B L E 4  Effects of supplement of dietary Ulva with various protein sources on the survival rate and growth performance of sea
cucumber (Apostichopus japonicus) for 8 weeks

Initial weight Survival

Diets (g) Final weight (g) Weight gain (%) SGR (%) FCR FI (g) rate (%)

U 9.34 ± 0.49a 14.55 ± 0.40 c 55.96 ± 3.96c 0.79 ± 0.05c 5.43 ± 0.30a 28.29 ± 1.06b 93.33

a bc c c a
F 9.75 ± 0.17 15.45 ± 0.82 58.34 ± 5.73 0.82 ± 0.06 5.21 ± 0.51 29.48 ± 0.52b 93.33
a bc c c a b
S 9.67 ± 0.14 15.49 ± 0.19 60.10 ± 0.33 0.84 ± 0.01 5.14 ± 0.09 29.86 ± 0.81 100
M 9.69 ± 0.17a 16.83 ± 0.74b 73.66 ± 4.69b 0.99 ± 0.05b 4.30 ± 0.23b 30.61 ± 0.81b 100
a a a a c a
B 9.60 ± 0.11 22.04 ± 0.23 129.60 ± 0.14 1.48 ± 0.01 2.93 ± 0.01 36.49 ± 0.33 100

Note: (1) Values are presented as mean ± SD. (2) Means in the same column with the different superscripts (a, b, c) are significantly different (p < .05).
(3) Weight gain: [(Final weight − Initial weight)/Initial weight] × 100%. (4) SGR (specific growth rate): [ln (Final weight (g)) − ln (Initial weight (g))/
day (s)] × 100%. (5) FCR (feed conversion ratio): feed intake (g)/(final weight (g) − initial weight (g)). (6) FI (feed intake): feed supplied (g dry weight).
(7) Survival rate: (Final number of sea cucumber/Initial number of sea cucumber) × 100%.

end of 8 weeks feeding trial. The best weight gain (35.20 ± 11.51%),
SGR (0.53  ±  0.15%) and FCR (5.35  ±  1.89) were found in U group
which were significantly better than other groups (p < .05).
Therefore, U. fasciata was selected as ingredient in the following ex-
perimental diet. The experimental diets were prepared containing
approximately 157.0–­172.1 g/kg crude protein and 15.0–­22.6 g/kg
lipid (Table 2). Results of growth performance of sea cucumber fed
different diets for 8 weeks were presented in Table 4. Sea cucumbers
fed with U diet exhibited the lowest weight gain (55.96  ±  3.96%),
SGR (0.79 ± 0.05%) and highest FCR (5.43 ± 0.30). There were no
significant differences among the growth performances of individu-
als fed with U, F and S diets (p > .05). The weight gain, SGR and FCR
of individuals fed with M and B diets showed significant improve-
ments compared with those of individuals fed with U, F and S diets
(p < .05). Moreover, sea cucumbers fed with B diet had significantly
better weight gain (129.60  ±  0.14%), SGR (1.48  ±  0.01%) and FCR
(2.93 ± 0.01) compared with individuals fed with M diet (p < .05).
(31.65 ± 2.43%) (p < .05). No significant difference was found among
the average attractiveness of U, F, S and M (p > .05).
3.3  |  Effects of dietary probiotic supplement
on the growth of Apostichopus japonicus
As shown in Table 6, the growth performance of sea cucumbers
was compared among four different conditions. Compared with the
reference condition (no probiotics supplement and no substrate),
the growth performance was not significantly affected in either di-
etary probiotics supplement alone or culturing with sand substrate
alone (p > .05). It was noted that dietary probiotics supplement to-
gether with culturing in sand environment could be beneficial for
growth of sea cucumbers. Under that condition, the weight gain
(30.63  ±  1.90%), SGR (0.45  ±  0.02%) and FCR (1.83  ±  0.10) were
significantly improved compared with those under other conditions
(p < .05).

3.2  |  Feeding preferences of Apostichopus japonicus

on various Ulva-­based diets 4  |  D I S C U S S I O N

As shown in Table 5, B was the most preferred diet with the high-
est average attractiveness (80.40 ± 11.48%). The relative attractive-
ness of B against other four diets exhibited above 65.08  ±  7.27%.
The average attractiveness of B was significantly higher than that
of U (49.13 ± 19.21%), F (45.10 ± 26.05%), S (43.33 ± 23.17%) and M
Previous studies have indicated that different size of sea cucumber
had varied seaweed choice and nutrition requirement (Xia, Yang, et al.,
2012; Xia, Zhao, et al., 2012). Our data suggested that U. fasciata was
the preferred diet for the growth of juvenile A. japonicus. Despite the
crude protein content of U. fasciata (157.0 g/kg) was lower than that of
LIAO et al.       5 |

G. tenuistipitata (171.0 g/kg). Apostichopus japonicus could have a good could reduce the content of crude fibre and might increase the content
digestibility of U. fasciata to compensate for its lower protein level. In of short-­chain peptide by means of acidic protease. Compared with
the second feeding experiment, four diets were prepared by replac- fish and soybean containing diets, BioMax-­containing diet improved
ing 60 per cent seaweed protein with the addition of fish, soybean its functional and nutritional properties as well as attractiveness for
and FSB (BioMax). Our results provided an optimized formula (B diet) A.  japonicus. Nowadays, studies evaluating the feeding preferences
for sea cucumber feed by the inclusion of U.  fasciata powder and of holothurians are still limited. Our data found that Lactobacillus sp.
BioMax at 1:1 weight ratio. As shown in Appendix 1, BioMax included fermented FSB could serve as an attractant and a growth enhancer
much higher ratio of free amino acids compared with soybean meal. simultaneously in feeding A. japonicus.
Moreover, the dietary inclusion of Lactobacillus sp.-­fermented BioMax Bacillus and Micrococcus species have been identified as probi-
otics and used to enhance the growth and immunity of aquaculture
species over the years (Abd El-­Rhman et al., 2009; Kuebutornye
TA B L E 5  The feeding attractiveness of five Ulva-­based diets for
Apostichopus japonicus et al., 2019). Particularly, dietary B.  subtilis significantly increased
the growth performance, innate immunity and disease resistance
Relative Average
of A. japonicus (Li et al., 2015; Sun et al., 2012; Zhang et al., 2010;
Diets Diets attractiveness (%) attractiveness (%)
Zhao et al., 2012). Moreover, 16S rRNA gene pyrosequencing of
U F 53.33 ± 9.18 49.13 ± 19.21b A.  japonicus revealed that genera Bacillus was present in foregut,
S 54.76 ± 4.12 hindgut and habitat surface sediment libraries, whereas genera
M 66.67 ± 8.25 Micrococcus were detected only in the sediment libraries (Gao et al.,
B 21.75 ± 6.14 2014). The Micrococcus bacteria in the sediment could be considered
F U 46.67 ± 9.18 45.10 ± 26.05b as a food source providing the host with essential nutrients or an
S 52.38 ± 4.12 activator of nutrient intake. Both intestinal autochthonous Bacillus
M 71.82 ± 13.38 and Micrococcus species were suggested as candidate strains for di-

B 9.52 ± 16.50
S U 45.24 ± 4.12
F 47.62 ± 4.12
M 68.25 ± 2.75
B 12.22 ± 10.72
M U 33.33 ± 8.25
F 28.17 ± 13.38
S 31.75 ± 2.75
B 33.33 ± 8.25
B U 78.25 ± 60.14
F 90.48 ± 16.50
S 87.78 ± 10.72
etary probiotics (Gao et al., 2014). Although the mixture of B. subtilis
b and M. luteus incorporated in diets did not significantly promote the
43.33 ± 23.17
growth of A. japonicus in bare tanks, our results showed a positive
effect on the dietary supplement of probiotics while sand was pro-
vided as a substrate in tanks. Sand or mud may be an essential part of
sea cucumber diets and substrate in tanks (Qiu et al., 2014; Robinson
31.65 ± 2.43b
et al., 2013). In rearing A. japonicus, the present study suggested that
sand may function as a digestive aid rather than a shelter.
Previous studies showed that A. japonicus fed solely macroalgae
or microalgae exhibited poor growth performance (Liu et al., 2009;
80.40 ± 11.48a Shi, Dong, Pei, et al., 2015; Shi et al., 2013). The inclusion of benthic
sediment or yellow soil in formulated diet is helpful to improve the
growth and production of sea cucumber (Liu et al., 2009; Shi, Dong,

M 65.08 ± 7.27 Wang, et al., 2015). However, the manufacture of benthic feed in

Note: (1) Values are presented as mean ± SD. (2) Means in the
same column with the different superscripts (a, b) are significantly
different (p < .05). (3) Attractiveness activity: (Attractive number/Test
number) × 100%.
practice is complex and time-­consuming. The present study sug-
gested a low-­cost and easily prepared diet for A. japonicus by using
the U. fasciata powder and FSB (BioMax) as ingredient. Comparing
with other studies using similar weight of A.  japonicus, our feed

TA B L E 6  Effects of the addition of probiotics in diet and sand as substrate on the survival rate and growth performance of sea cucumber
(Apostichopus japonicus) for 8 weeks

Initial weight Weight gain Survival

Probiotics/Substrate (g) Final weight (g) (%) SGR (%) FCR FI (g) (%)

−/− 30.18 ± 0.51a 33.52 ± 0.11b 11.10 ± 2.26b 0.18 ± 0.03b 5.24 ± 1.08a 17.20 ± 0.35a 100

a ab b b a
−/+ 32.23 ± 2.67 35.68 ± 1.93 10.83 ± 3.20 0.17 ± 0.05 5.46 ± 1.54 18.26 ± 1.27a 100
a ab b b ab a
+/− 31.60 ± 2.32 36.35 ± 3.21 14.99 ± 1.73 0.23 ± 0.03 3.60 ± 0.64 16.85 ± 0.17 100
+/+ 32.00 ± 2.04a 41.82 ± 3.27a 30.63 ± 1.90a 0.45 ± 0.02a 1.83 ± 0.10 b 17.89 ± 1.28a 100

Note: (1) Values are presented as mean ± SD. (2) Means in the same column with the different superscripts (a, b) are significantly different (p < .05).
(3) Weight gain: [(Final weight − Initial weight)/Initial weight] × 100%. (4) SGR (specific growth rate): [ln (Final weight (g)) − ln (Initial weight (g))/
day (s)] × 100%. (5) FCR (feed conversion ratio): feed intake (g)/(final weight (g) − initial weight (g)). (6) FI (feed intake): feed supplied (g dry
weight). (7) Survival rate: (Final number of sea cucumber/Initial number of sea cucumber) × 100%.
 |
6      LIAO et al.
formulation fit the nutritional requirements of small juvenile sea cu-
cumber with the high SGR value of 1.48%, which was better than
0.85% reported from other study using complex feed formulation
including algae powder, fish meal, sea mud, soybean meal and syn-
biotic of Bacillus lincheniformis and alginate oligosaccharides (Wang,
Sun, et al., 2017). In addition, the system of dietary probiotics with
sand bottom had a comparable SGR of 0.45% to SGR of 0.23%–­
0.99% reported from the previous study feeding the middle size of
juvenile A. japonicas in tank culture by complex diets supplemented
with graded levels of hydrolysed soybean meal (Song et al., 2016).
Therefore, our simple feed formulation and culturing system can be
easily applied to sea cucumber aquaculture in Taiwan.
AC K N OW L E D G E M E N T
The second author thanks the International Cooperation and
Development Fund (Taiwan ICDF) for granting the scholarship of
the Master Program in Aquaculture Technology and Management.
C O N FL I C T O F I N T E R E S T
The authors declare that they have no conflicts of interest.
ORCID
Fan-­Hua Nan  https://orcid.org/0000-0003-4133-7171
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(Selenka). Aquaculture, 338, 304–­3 08. https://doi.org/10.1016/j. Lysine 49.4 1619.3
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Analysis was conducted according to China Standards System
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marcusii DB11 supplementation on the growth, immune response,
and expression of immune-­ related genes in coelomocytes and