Aquacultura Indonesiana (2013) 14 (2): 85-97

ISSN 0216-0749

Effects of different diet regimes on development of Gill and Rostrum spines of Pacific
white shrimp Litopenaeus vannamei
Romi Novriadi
Balai Budidaya Laut Batam, Direktorat Jenderal Perikanan Budidaya,
Kementerian Kelautan dan Perikanan, Jl. Raya Barelang, PO BOX 60 Sekupang,
Batam 29422, Kepulauan Riau, Indonesia
Telphone: (0778) 7027623 7027624, Faksimile: (0778) 3582557

Abstract
Romi Novriadi. 2013. Effects of different diet regimes on development of gill and rostrum spines of
pacific white shrimp Litopenaeus vannamei. Aquacultura Indonesiana, 14 (2): 85-97. The effect of partial
replacement of Artemia nauplii with artificial diets were evaluated on gill and rostrum spines development as
well as the quality of white shrimp (Litopenaeus vannamei, Boone) post larvae (PL). The traeatments were: (1)
live food control Artemia nauplii, (2) 65% replacement of Artemia with artificial diet, and (3) 85% replacement
of Artemia with artificial diet. All treatments were also compared to the L. vannamei PL generated from
commercial hatcheries. Instar II Artemia nauplii were cultured as a starter live food and Frippak microencapsulated
feeds were provided as the artificial diet to L. vannamei. A significantly better quality, gill and rostrum spines
development was achieved by post larvae (from PL1 to PL 5) fed with live Artemia or the 65% replacement of
Artemia in comparison to 85% replacement of Artemia. Interestingly, even had the lowest quality, PL fed with
85% replacement of Artemia nauplii still recorded a superior quality compared to commercial hatchery products
at the same PL stages, suggesting that the nutrition standard performed at the commercial hatchery in the
sampling area does not support the optimal growth of gill and rostrum spines as well as the quality of L. vannamei
at post larva stages. Additionally, the findings are important in aquaculture where the appropriate concentrations of
micro particles are also able to demonstrate a better performance in L. vannamei post larvae quality and this may
contribute to the efficiencies towards a reduction of Artemia nauplii cost.
Keywords : L. vannamei; Diet regime; Gill; Rostrum spines; Larva index quality

Abstrak
Dampak penggantian sebahagian pakan naupli Artemia dengan pakan buatan dievaluasi dengan
melakukan analisa terhadap perkembangan insang dan rostrum spines serta kualitas post larva udang Vannamei
(Litopenaeus vannamei, Boone). Perlakuan terdiri atas: (1) pemberian naupli Artemia, (2) 65% penggantian
Artemia dengan pakan buatan, dan (3) 85% penggantian Artemia dengan pakan buatan. Seluruh perlakuan juga
dibandingkan dengan PL udang Vannamei yang dihasilkan oleh panti benih komersial. Fase instar II naupli
Artemia digunakan sebagai pakan hidup dan Frippak Microencapsulated diet digunakan sebagai pakan buatan
untuk udang L. vannamei. Sebuah kualitas, perkembangan insang dan rostrum spines yang lebih baik secara
nyata dicapai oleh post larva (dari PL1 ke PL5) yang diberi pakan Artemia atau 65% penggantian Artemia jika
dibandingkan dengan 85% penggantian Artemia. Menariknya, walaupun memiliki kualitas yang lebih rendah,
PL yang diberi pakan dengan 85% penggantian Artemia masih memberikan kualitas yang lebih baik jika
dibandingkan dengan produk panti benih komersil pada tahapan PL yang sama, menunjukkan bahwa standar
pemberian pakan yang dilakukan oleh panti benih di wilayah pengambilan sampel tidak mendukung
perkembangan insang dan rostrum spines serta kualitas udang L. vannamei pada tahapan post larva. Selain itu,
penelitian ini penting untuk kegiatan budidaya dimana konsentrasi penggunaan pakan buatan yang tepat juga
mampu menghasilkan pertumbuhan, osmoregulasi dan kemampuan detoksifikasi yang lebih baik dan dapat
berkontribusi pada efisiensi terhadap pengurangan biaya penggunaan Artemia nauplii.
Kata kunci : L. vannamei; Pemberian pakan; Insang; Rostrum spines; Indeks kualitas larva

Introduction
Pacific white shrimp (L. vannamei,
Boone), is an economically important species that
is widely cultured not only in their native regions
at western pacific coast of Latin America but also
in the expansive Asia (Liao and Chien, 2011).
However, the success story of L. vannamei farming

has always been crippled by the lack of high-quality

post larvae. One of the major factors hampering the
quality of hatchery-reared post larvae is nutrition,
especially during the critical periode where there
is an extensive morphogenesis in the digestive
system occurs at the first 10 days of post larvae
(Lovett and Felder, 1989). The behavioural changes
from herbivorous (filter feeders) to carnivorous

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Aquacultura Indonesiana, Vol. 14, No.2, Agustus 2013 :85-97

(hunters), which occur due to the life cycle

development of shrimps require appropriate
nutrition and correct prey size (Lavens and
Sorgeloos, 1996). Shrimp will be more herbivorous
at zoea stages and therefore require microalgae such
as diatoms Chaetoceros spp., Thalassiosira spp.
and Skeletonema spp. (Cook and Murphy, 1966;
Emmerson, 1980; Martins et al., 2006; Soares et al.,
2006), whereas at the mysis and postlarva stage,
penaeid shrimp becomes more carnivorous and
require zooplankton (Lavens and Sorgeloos, 1996).
Freshly hatched Artemia nauplii are the
most widely used as live feed for early stage
crustacean larvae due to its high nutritional
quality and ease of use (Sorgeloos et al., 1998).
The suitability of Artemia nauplii as live feed in
crustacean larviculture is also supported by the
presence of the fatty acid 20:5 (-3)
eicosapentaenoic acid or EPA. Artemia nauplii is
usually given in live condition to give an
optimum feeding regime especially on the mysis
and early postlarval stages (Mc Fey and Fox,
1983). In addition, several forms of Artemia have
also been used in most penaeid hatcheries to
induce an optimal growth: i.e. heat killed nauplii,
frozen and blended Artemia (Wilkenfeld, 1984;
Wouters and Van Horenbeeck, 2003; Juarez et
al., 2010). However, study from Garca-Ortega et
al. (1995) indicated that heat killed nauplii have a
negative impact to the protein content and enzyme
activity of Artemia, where at 60C more than 50%
of the enzymatic activity was decreased and around
30% of the protein was denatured.
Significant increase in the operational
cost of shrimp hatcheries has been considered as
a major contsraint in the use of live nauplii.
Recent research have include the use of rotifers
Brachionus plicatilis (Emmerson, 1980; Naessens
et al., 1995), nematodes Panagrellus redivivus
(Wilkenfeld, 1984; Biedenbach et al., 1989; Focken
et al., 2006) and extending the use of algal food for
the late mysis and post larval stages as a potential
alternatives for substitute live feed Artemia
(Gopalakrishnan,
1976). Moreover,
microparticulate and micro-encapsulated formulated diets
can also be used to reduce the amount of Artemia
cysts (Galgani and Aquacop, 1988; Wouters et al.,
2009)
Many scientists have studied the effect of
the supplement diets on survival, growth or total
length of penaeid shrimps (Biedenbach et al., 1989;
Hirata et al., 1985; Cobo, 2013; Naessens et
al.,1995). Moreover, Brito et al. (2001) have stated
that partial substitution (50%) of Artemia by
artificial diet and the use of algae beyond the first
post-larval stage have a benefit impact to the

growth and nutritional state of L. vannamei.

However, In the present study, we more focus on
the effect of partial replacement of Artemia with
different percentage of Frippak microencapsulated
diet to the development of gill and rostrum spines
of L. vannamei. In addition, examinations of post
larvae quality, including: growth, salinity stress test,
dry weight and wet weight was also performed
together with comparative study of post larva
quality generated from several commercial
hatcheries.

Material and Methods

Experiment Animals
Larvae
of
Pacific
white
shrimp
(L. vannamei, Boone) were used as an experimental
animal and reared in a 175 L experimental tank
with density of 175 shrimp L. vannamei/tank.
The water quality parameters, the photoperiod,
temperature and the feeding regime were adjusted.
Water was exchanged at a rate of aproximately 50%
/day after removing waste and uneaten feed by
siphoning. NH4-N, NO2-N, and NO3-N levels were
maintained below 0.2, 0.1 and 5 mg/L, respectively.
Gentle aeration was applied in all rearing tank to
maintain the dissolved oxygen concentration above
5 mg/L. The photoperiod was set at 12:00 hours
light at an intensity of 900-1000 lx with fluorescent
lamps at the water surface. Newly hatched Artemia
(SepArt, New generation High 5 Artemia, INVE
aquaculture) were used as live food and
administered via the L. vannamei culture water
from PL 1 to PL 5. The Artemia dosage was
calculated based on the feeding level and the
number of shrimp larvae in the tank and split over
two feedings at 07.00 hours and 16.00 hours.

Experimental Design
The experimental set-up were performed in
spherical tank containing 175 L saline water
(30 ppt). The spherical tank were placed in
hatchery containing water maintained at 291oC
using thermostatic heater. A lamp system was
installed to provide around 900-1000 lx at
the water surface. The tank were supplied with
gentle aeration to ensure the dissolved oxygen in
the rearing water of L. vannamei was always over
5 mg/L. Each tank were stocked with batches of
175 nauplii of L. vannamei. In the first trial,
stocking was performed using a volumetric
method. Even though the L. vannamei nauplii at
this stage still deplete internal reserves and do not
feed on algae, 20 algal cells/mL of Chaetoceros
calcitrans was added to guarantee the feed supply

86Hak cipta oleh Masyarakat Akuakultur Indonesia 2013

Effects of different diet regimes on development of gill and rostrum spines of pacific white shrimp Litopenaeus vannamei
(Romi Novriadi)

once the animals reached the protozoea stage.

Feeds was provided 6 times per day and the number

of Chaetoceros calcitrans was counting by

Haemocytometer.

Table 1. Type and amount of feed provided for each successive stage of larval development of L. vannamei
Stage

Live food control

65% substitution

85% substitution

Nauplii 5
Protozoea 1-2

C.calcitrans 20 cells/L
C.calcitrans 50 cells/L

C.calcitrans 20 cells/l
C.calcitrans 50 cells/L

C.calcitrans 20 cells/l
C.calcitrans 50 cells/L

Protozoea 3
Mysis 3

C.calcitrans 50 cells/L
Artemia nauplii 2-10/mL

PL1 PL5

Artemia nauplii 5-10/mL

C.calcitrans 50 cells/L
65% substitution of Artemia
nauplii with Frippak
65% substitution of Artemia
nauplii with Frippak

C.calcitrans 50 cells/L
85% substitution of Artemia
nauplii with Frippak
85% substitution of Artemia
nauplii with Frippak

In our experiments, an open clear water

system was used with a daily water exchange of
50%. During water exchange, the remaining (heat
killed) Artemia and waste from the previous day
were removed by siphoning. This operation was
carried out with great care to avoid loss of larvae.
Feeding was done after water exchange.
Three different treatments with four
replicates each were tested (Table 1): The life
food control (Artemia only) and two substitution
of Artemia with 65% and 85% commercial feeds,
respectively. In the first stages, from Nauplii 5 to
Protozoea 2, all treatments were fed with
Chaetoceros calcitrans. From Protozoea 3 to
Post larvae 5, Artemia nauplii substituted with
commercial feeds (INVE Frippak) were given.
This kind of feeding strategy had been used as
Artemia treatment in former experiment at INVE
SCRD-Thailand.
The
experiments
were
terminated when 90% of the L. vannamei
treatment in one replicate reached the first PL5
stages. The larvae and post larvae were measured
in the frame of post larva quality analysis.

Larval Quality Analysis

Determination of Gill and Rostrum Spines
Development
Artemia nauplii were substituted with
INVE Frippak commercial feeds based on
the feeding level and the number of shrimp larvae
in the tank. Gill and rostrum spines development
in each treatment was detected under light
microscope equipped with a Canon EOS digital
camera connected to a PC. The images were
compared with gill and rostrum development at
the same post larvae stages from commercial
hatcheries in Chonburi-Thailand.
Post Larva Quality

Post larva quality was checked at every

stage of PL by analyzing the osmotic stress, dry
weight, wet weight, and average length among
the treatments. Size of the Post Larvae was
measured according to Kitani (1986) from the
base of the antennal flagellum to the telson, and
the mean of total length and coefficient of
variation were calculated for each treatment and
PL stages of sampling. To evaluate the post larvae
survival to osmotic stress, a simple stress test was
performed with some modification. Each treatment
was left for 60 minutes at different range salinity,
namely: 1, 2 and 3 ppt, respectively and the number
of survivors was counted after the specified time.
Dry weight analysis were performed by drying 1 g
of post larvae in the oven for 24 hours at 60oC and
measured using analytical balance. Wet weight
obtained by measuring 1 g and dividing by the
number of existing post larvae.
Statistics
The results are presented as mean values
followed by the standard deviation and the
percentage data were arcsine transformed for
statistical comparisons to satisfy normal
distribution and homoscedasticity requirements.
Data were then subjected to one way analysis of
variances followed by Tukeys multiple
comparison range using the statistical software
SPSS version 21.0 to determine significant
differences among treatments. All significance level
of the statistical analysis was set at P<0.05.
Results
Gill and Rostrum Spines Development
Microscopic analysis showed that the
development of gill and rostrum spines varies for
each treatment. At PL 1 stages, the control and
Frippak 65% have 3 dorsal rostral spines plus one

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Aquacultura Indonesiana, Vol. 14, No.2, Agustus 2013 :85-97

growth indications of spine between the first spines

(at the edge) and the second spines. Frippak 85%
treatment only have 3 dorsal rostral spines without
any indication of further spines growth. At PL 2
stages, Live food control and Frippak 65% have 4
dorsal rostral spines each, while the Frippak 85%
still only have 3 dorsal rostral spines with new
indication of spines growth. At PL 3 stages, Live
food control and Frippak 65% have 4 dorsal spines
each with the addition of setae and the second
spines. However, at Frippak 85% there were

Live food control

variations in the number of spines (3 and 4 dorsal

spines) with and without setae. At PL 4 stages, Live
food control and Frippak 65% have 4 dorsal spines
each with 2-3 setae at the second dorsal spines, while
the Frippak 85% also have 4 dorsal spines but only
have 1-2 setae at the second dorsal spines. Finally at

PL 5, live food control have 5 dorsal rostral spines,

while Frippak 85% have 4 dorsal rostral spines. The
difference in rostrum spines became more and more
pronounced towards the end of the experiment
(Figure 2)

65% substitution

85% substitution

PL 1

PL 2

PL 3

PL 4

PL 5

(1)

(2)

(3)

Figure 1. The development of gill at PL1 to PL5 of L.vannamei . Life food control indicates the effect of
Artemia administration to the gill development (1), 65% substitution indicates the effect of 65%
substitution of Artemia by artificial diet to the gill development (2) and 85% substitution indicates the
effect of 85% substitution of Artemia by artificial diet to the gill development (3)

88Hak cipta oleh Masyarakat Akuakultur Indonesia 2013

Effects of different diet regimes on development of gill and rostrum spines of pacific white shrimp Litopenaeus vannamei
(Romi Novriadi)

Live food control

65% substitution

85% substitution

PL 1

PL 2

PL 3

PL 4

PL 5

Figure 2. The development of rostrum spines at PL1 to PL5 of L.vannamei. Life food control indicates the effect
of Artemia administration to the rostrum development (1), 65% substitution indicates the effect of
65% substitution of Artemia by artificial diet to the rostrum development (2) and 85% substitution
indicates the effect of 85% substitution of Artemia by artificial diet to the rostrum development (3)

The treatments with Artemia (live food

control) and Frippak 65% at PL 2 indicate the
presence of 2-3 branched at their gill, while
Frippak 85% remain unbranched. At PL 3, In
general, live food control and Frippak 65% have
5 branches in their gill, while Frippak 85% varies
from 2 to 5 branches. At PL 4, live food control
and Frippak 65% indicated 7 to 9 branches, while
Frippak 85% varied from 5 to 7 branches.
Finally, at PL 5, live food control and Frippak
65% showed 10 to 11 branches in each of their

gill, while the branches in Frippak 85% varied

between 3 to 9.
Post Larva Quality Analysis
1 Average length and dry weight
Larvae fed with Artemia nauplii were
significantly longer (P<0.05) in average of length
at PL 5 (0.9 cm) followed by 65% substitution
treatment (0.8 cm), and this group was
significantly different with other treatment

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Aquacultura Indonesiana, Vol. 14, No.2, Agustus 2013 :85-97

(P<0.05). 85% substitution of Artemia nauplii

resulted in the shortest of average length at the
end of experiment (PL5) (Figure 3). Post larvae
of L.vannamei fed with Artemia nauplii (Live
food control) were significantly (P<0.05) heavier
in terms of dry weight (0.218 mg) than those fed

with other treatments. Feeding at 65%

substitution were significantly (P<0.05) heavier
in dry weight than 85% substitution. The dry
weight variation trends were observed from PL3
stages.

Figure 3. Average length of L. vannamei from PL1 to PL 5, fed with Artemia naulii (LFC = Live Food Control),
65% S and 85% S (S= substitution of Artemia nauplii with Frippak INVE Commercial feed),
respectively. Values are averageSE, n=4. Different letters denote significant differences (P<0.05).
Different types of superscript denote different comparison.

Figure 4. Average dry weight of L. vannamei post larvae from PL1 to PL 5 fed with Artemia naulii
(LFC = Live Food Control), 65% S and 85% S (S= substitution of Artemia nauplii with Frippak
INVE Commercial feed), respectively. Values are average SE, n=4. Different letters denote
significant differences (P<0.05). Different types of superscript denote different comparison.

2. Salinity stress analysis

Post larvae of L.vannamei are maintained
in saline water (30 ppt) at 30 0,5oC before the
salinity shock treatment. All the treatments (live
food control, 65% substitution and 85%

substitution) were exposed to three low salinity

solution (1 ppt, 2 ppt and 3 ppt) at ambient
temperature and the survival was determined
after 1 h. As much as twenty post larvae were
used per trial at every post larvae stages.

90Hak cipta oleh Masyarakat Akuakultur Indonesia 2013

Effects of different diet regimes on development of gill and rostrum spines of pacific white shrimp Litopenaeus vannamei
(Romi Novriadi)

Figure 5.

Mortality (%) of PL 1 to PL 5 L. vannamei post larvae fed Artemia nauplii with or without
commercial feed. In the experiment, LFC: Live food control (only fed with Artemia nauplii), 65%
S : 65% substitution of Artemia nauplii with commercial feeds, and 85% S: 85% substitution of
Artemia nauplii with commercial feeds. All the treatments exposed to 1 ppt solution. Values are
averageSE, n=3, Different letters denote significant differences (P<0.05).

The results presented in Figure 5 showed

that a significant low percentage mortality
(P<0.05) was observed in L. vannamei post
larvae fed with Artemia nauplii exposed to 1 ppt
salinity solution in comparison to 85% partial
substitution of Artemia with microencapsulated
diet at every stage of post larvae development.
However, there is no significant difference in
terms of percentage mortality was observed

between post larvae of L. vannamei fed with

Artemia in comparison to post larvae that receive
65% partial replacement of Artemia.
Similar trend was also noted at 2 ppt and
3 ppt treatments solution (Figure 6 and 7). The
treatments with both live food control and 65%
substitution resulted in the lowest mortality
percentage in comparison to 85% partial
replacement of Artemia nauplii (P<0.05)

Figure 6. Mortality (%) of PL1 to PL5 L.vannamei post larvae fed Artemia nauplii with or without commercial
feed. in experiment, LFC : Live food control (only fed with Artemia nauplii), 65% S : 65%
substitution of Artemia nauplii with commercial feeds, and 85% S : 85% substitution of Artemia
nauplii with commercial feeds. All the treatments exposed to 2 ppt solution. Values are averageSE,
n=3, Different letters denote significant differences (P<0.05).

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Aquacultura Indonesiana, Vol. 14, No.2, Agustus 2013 :85-97

Figure 7. Mortality (%) of PL1 to PL5 L.vannamei post larvae fed Artemia nauplii with or without commercial
feed. in experiment, LFC : Live food control (only fed with Artemia nauplii), 65% S : 65%
substitution of Artemia nauplii with commercial feeds, and 85% S : 85% substitution of Artemia
nauplii with commercial feeds. All the treatments exposed to 3 ppt solution. Values are average SE,
n=3, Different letters denote significant differences (P<0.05).

3. Post larva quality

commercial hatcheries.

comparison

with

12 shrimp hatcheries was visited in the

frame of sampling campaign to obtain a
comparative data between post larva quality

generated from trial work with post larva quality

from commercial hatchery. Sampling was carried
out using local equipment and sample were kept
on plastic filled with oxygen during
transportation.
Observation Parameter

Hatchery Farm

Type of shrimp

Feeding scheme

wet weight Dry weight average of

PL stages

Prom-Phong Hatchery Penaeus Vannamei

Zoea : C. Calcitrans + artificial diet+ heat killed Artemia

( gr)

( gr )

mortality with osmotic stress (1 H)

length (cm)

1 g/L

2 g/L

3 g/L

Total

Type of Artemia

SR

water quality (at sampling)

T (C)

sal ( g/L)

PL 6

0,0013624 0,000326

0,71032

10

Brand "white bear"

50%

25

30

PL 8

0,00223214 0,00047

0,91085

Brand "Galaxy"

60%

27

25 (rainy day)

PL 7

0,00174216 0,0004546

0,8326

Brand " Swan GSL"

50-60 %

30

30

PL 6

0,00110865 0,000276

0,7116

Brand "Eagle" Artemia

50%

30

30

PL 12

0,00649351 0,0015633

0,9951

Brand " Aqua " AQP

50%

30

30

PL 8

0,0037594 0,00053867

0,9817

Brand "Galaxy"

50%

29

20

Mysis : C. Calcitrans + artificial diet+ heat killed Artemia

PL 1-12 : Daphnia + Artemia + artificial diet
Pichitpon Farm

Penaeus Vannamei

Zoea : C. Calcitrans + artificial diet+ heat killed Artemia

Mysis : C. Calcitrans + artificial diet+ heat killed Artemia
PL 1-12 : Artemia + artificial diet+ Black flakes

Lab-Inter2 Farm

Penaeus Vannamei

Nauplii : C. Calcitrans
Zoea : C. Calcitrans + artificial diet+ heat killed Artemia
Mysis : C. Calcitrans + artificial diet+ heat killed Artemia
PL 1-12 : Artemia + artificial diet+ Black flakes

55 Farm

Penaeus Vannamei

Nauplii : C. Calcitrans
Zoea : C. Calcitrans + artificial diet+ heat killed Artemia
Mysis : C. Calcitrans + artificial diet+ heat killed Artemia
PL 1-12 : Artemia + Daphnia + artificial diet+ Black flakes

Phanh Tib Farm

Penaeus Vannamei

Nauplii : C. Calcitrans
Zoea : C. Calcitrans + Spirulina + heat killed Artemia
Mysis : C. Calcitrans + artificial diet+ heat killed Artemia
PL 1-12 : Artemia +artificial diet

Chaleam Farm 1

Penaeus Vannamei

This hatchery is only as a transit step at PL 8-12

Artemia only applied when packaging to avoid the stress condition in shrimp

Chanchira Farm 2

Penaeus Monodon

Mysis to PL 1 : heat killed artemia (Instar-1)

PL 2

0,0015008 0,00021067

0,704

20

20

20

60

Brand M.T.I

> 80 %

30

30

Chanchira Farm 1

Penaeus Monodon

Mysis to PL 1 : heat killed artemia (Instar-1)

PL 3

0,0013298 0,00019933

0,7548

20

20

20

60

Brand M.T.I

> 80 %

30

30

Chanchira Farm

Penaeus Vannamei

Mysis to PL 1 : heat killed artemia (Instar-1)

PL 1-12 : Artemia +artificial diet

PL 3

0,00065934 0,000096

0,4914

Brand M.T.I

60%

30

30

Pasit Farm 1

Penaeus Monodon

Nauplii : C. calcitrans

PL 8

0,0117375 0,0002967

0,70165

20

20

20

60

Brand " Premium"

> 80 %

30

30

60-70 %

30

30

10-50%

30

30

Mysis to Zoea 1 : heat killed Artemia (instar 1)

Red and green package

Zoea 2 to PL 4 : Artemia
PL 5 onwards : Daphnia
Pasit Farm

Penaeus Vannamei

Nauplii : C. calcitrans

PL 3

0,00105568 0,000099

0,5973

10

Mysis to Zoea 1 : heat killed Artemia (instar 1)

Brand " Premium"

Red and green package

Zoea 2 to PL 4 : Artemia
PL 5 onwards : Daphnia
Ma win Farm

Penaeus Vannamei

Zoea I-III : Spirulina powder + Chaetoceros

PL 10

0,00325733 0,00043313

0,8103

zoea II they applied the heat killed artemia

PL 1 onwards : Artemia + artificial diet
sometimes mix with Vitamin

Table 2. Summary of sampling campaign in 12 commercial hatcheries

92Hak cipta oleh Masyarakat Akuakultur Indonesia 2013

Brand " M.C.C "

Effects of different diet regimes on development of gill and rostrum spines of pacific white shrimp Litopenaeus vannamei
(Romi Novriadi)

There were two commercial hatcheries

that could be compared due to the availability of
PL size based on the observed post larva stage,
namely: Chanchira Farm (PL 3 of L.vannamei)
and Pasit Farm (PL 3 of L.vannamei). Based on
the post larva quality and microscopic analysis,
PL from commercial hatcheries appeared to grow
slightly lower in terms of dry weight and average
length even when compared to Frippak 85%
substitution that generated from our experiment.
Percentage mortality in 1 g L-1 solution also

slightly higher if compared with live food control

and 65% substitution treatments but equal to the
mortality number of 85% substitution treatment.
Similar thing was also seen in the 2 ppt and 3 ppt
solution treatments. No replicate measurements
are available for commercial hatcheries sample to
enable statistical evaluation mainly due to the
limitation of sample. Gill and rostrum
development was also slower for both
commercial hatcheries sample.

Figure 7. Rostrum spines (A) and gill (B) development of L.vannamei at PL 3 in Pasit Farm and rostrum spines
(C) and gill (D) development of L.vannamei at PL 3 in Chanchira farm.

Discussion
In this study, we observed the effect of
different diet regimes on the pacific white shrimp
L.vannamei post larvae performance from PL1
to PL5 and on the rostrum spines and gill
development. It is evident that the administration
of live Artemia nauplii from Mysis to Post larvae
5 resulted in better gill and rostrum spines
development (Figure 1 and 2). Moreover, These
results showed no significant differences between
control and 65% partial replacement of Artemia
in respect to the gill and rostrum spines
development along with the quality of post
larvae. In contrast, 85% replacement of Artemia
nauplii resulting in the lowest quality in
comparison to other treatments. In term of gill
development, as an euryhaline tropical shrimp,
L. vannamei highly depend on gill, mainly for
regulation of their osmoregulatory capacity and

tolerance of salinity fluctuations in the water

(Wickins and Lee, 2002). Salinity is one of the
most important abiotic factors for marine and
estuarine organisms, which affect their growth
and survival (Saoud et al., 2003; Buranajitpirom
et al., 2010; Davis et al., 2005; Pqueux, 1995;
Fielder et al., 2001; Atwood et al., 2003 and
Kumlu et al., 1999). Moreover, mass mortalities
in grow out ponds of penaeid shrimp is often
related to the fluctuations of salinity levels and
less observed in smaller sized organisms because
they are better osmoregulators (Vargas-Albores
and Ochoa, 1992).
An interesting observation was that the
treatments also resulted in similar response to the
rostrum spines development (Figure 2). Overall,
85% substitution of Artemia nauplii resulted to a
slower development of gill and rostrum spines.
However, further studies are required to
determine which of the diet (HUFA,

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Aquacultura Indonesiana, Vol. 14, No.2, Agustus 2013 :85-97

macronutrient
and
micronutrient)
or
environmental factors is responsible, or whether
they act synergistically to induce the
development of gill and rostrum spines at post
larva stages of L. vannamei.
Another important feature arising from
this trial is the impact of different feeding regime
to the growth and survival of L. vannamei post
larvae when tested with osmotic stress factor.
Importantly, we found that control and 65%
substitution of Artemia nauplii, which had a
better development of gill and rostrum spines had
essentially the same survival levels (Figure 5)
when exposed to 1, 2 and 3 ppt solution from 30
ppt for 1 hour. Their survival was significantly
higher in comparison to 85% substitution
(P<0.05). Dietary manipulations have important
roles to improve the osmoregulatory abilities of
culture L. vannamei subjected to osmotically
stressful conditions which in turn is expected to
increase the quality and productivity of
aquaculture production (Romana and Zeng,
2012). This mainly due to the osmoregulation
process require energy that are sourced from
protein (Setiarto et al., 2004; Rosas et al.,
1999 and Silvia et al., 2004) and/or lipids
(Palacios et al., 2004; Luvizotto-Santos et al.,
2003; Lemos et al., 2001 and Sang and Fotedar.
2004). Therefore it seems reasonable to assume
that providing an easy digestible live food and
readily available energy supply with optimal
substitution concentration could effectively
improve the survival of L. vannamei in extreme
change of environment salinity.
In addition to the importance of diet,
every stage of L. vannamei responded similarly
to several feed and fish meals in
microencapsulated diet (Cousin et al., 1993). If
there is a difference in digestibility, potential
exist appear between larvae and postlarvae.
Figure 3 showed that from PL 1 to PL4, there is
no significant difference (P<0.05) in average
length between live food control and 65%
substitution. However, both of the treatments
(LFC and 65% substitution) have a significant
differences (P<0.05) compared to 85%
substitution. Furthermore, in the end of
observation (PL 5), LFC (live food control) had a
larger average length (P<0.05) than 65%
substitution. From PL 1 to PL 5, 85% substitution
resulted in poor growth in terms of average
length. The trials emphasized that native protein
seems to be better hydrolyzed than the processed
one (Zwilling et al., 1981). This is in agreement
with studies of Cuzona et al. (2004), who

reported that more digestible nutrients absorbed

promotes more growth to the cultured aquatic
animals. The provision of feed by using Artemia
nauplii also appears to provide a better growth, as
the post larvae stage had a significantly heavier
(P<0.05) dry weight compared to 65% and 85%
substitution of Artemia nauplii. This difference is
very evident from PL 3 to PL 5 and 65%
substitution still have a better performance than
the 85% substitution.
In comparison of our PL stages with
those from several commercial hatcheries, the
quality of L. vannamei post larvae coming from
commercial hatcheries was not as expected. The
gill and rostrum spines development in both
hatcheries: Pasit and Chanchira farm was slower
in terms of gill lamellae branch and rostrum
spines. Microscopic analysis show that the PL 3
stages from Pasit and Chanchira farm is nearly
equal to PL 3 of 85% substitution and PL 2 of
life food control respectively. In general, the
quality performance of PL from both hatcheries
were still lower than life food control and 65%
substitution. We note that these conditions were
caused by inadequate nutrition in their Artemia
cyst due to the diversity of cyst products in the
market and improper Artemia hatching procedure
and feeding regime.
In the present study, appropriate
concentrations of microencapsulated diets can
also provide a better growth and performance of
L. vannamei post larvae as well as gill and
rostrum spines development in comparison to
post larvae that fed with Artemia nauplii. The
results showed that an appropriate concentrations
of micro particles in the diet will also make the
feed more digestible and provide adequate
nutrition to induce gill and rostrum development
resulting in beneficial effects on L. vannamei
post larvae performance.
Conclusion
Artemia nauplii had good dietary and
promote the optimal development of gill and
rostrum spines for Pacific white shrimp
(L. vannamei, Boone) at post larvae stages. The
constant
dietary
availability
on
microencapsulated diet in Frippak show no
significant differences in gill and rostrum spines
development compared to Artemia nauplii live
food control only when 65% of Artemia was
substituted with this diet, while the differences
become significant when the live food
substitution increase up to 85%. Gill

94Hak cipta oleh Masyarakat Akuakultur Indonesia 2013

Effects of different diet regimes on development of gill and rostrum spines of pacific white shrimp Litopenaeus vannamei
(Romi Novriadi)

development is very important and could induce

the protection against osmoregulatory stress
especially during the critical periode on their life
cycle. Surprisingly, The quality of post larvae
generated from this study was better than the post
larvae coming from commercial hatchery. The
quality varied in L. vannamei post larvae in
accordance to average length, dry weight,
osmotic stress, gill and rostrum spines
development. The apparent capacity of Artemia
nauplii to promote better quality of post larvae
production offers an interesting reality that the
substitution of Artemia with microencapsulated
diet in early development of L. vannamei remains
a challenge. To our knowledge, this study,
reports for the first time the effect of substituting
Artemia nauplii on the development of gill and
rostrum spines of L. vannamei, mainly at post
larva stages from PL1 to PL5.
Acknowledgements
The author acknowledges the grant
support by De Vlaamse Interuniversitaire Raad
(VLIR) Belgium for the Internship program and
the technical assistance by Patipon Srianek
(Thailand).
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