Effect of Spirulina platensis on the productive performance of ...
嚜澠nternational Journal of Agricultural Science Research Vol. 2(9), pp. 273-278, September 2013
Available online at
ISSN 2327-3321 ?2013 Academe Research Journals
Full Length Research Paper
Effect of Spirulina platensis on the productive
performance of Litopenaeus vannamei (Boone, 1931)
shrimp
Ruth Gomes de Figueiredo GADELHA 1*, Jo?o Andrade da SILVA2, Neiva Maria de Almeida3
and Ana Herm赤nia Andrade e SILVA4
Programa de P車s-Gradua??o em Ci那ncia e Tecnologia de Alimentos; Universidade Federal da Para赤ba; 58051-900;
Jo?o Pessoa, PB - Brasil.
Accepted 26 August, 2013
Supplementation with Spirulina platensis has shown excellent results on growth when used as a food
source in aquatic animals. Therefore, the present study aimed to evaluate the effect of S. platensis in
diets with different percentages in the productive performance of Litopenaeus vannamei marine shrimp.
Five experimental diets were prepared (0, 10, 20, 30 and 40%) with all isonitrogenous (35%) and
isocaloric (3400 kcal) tested in a completely randomized design consisting of five treatments and three
replications. Juveniles were used with initial weight of 1.42 g fed for 45 days ad libitum three times a
day with the experimental diets and a commercial feed with the same protein percentage. Analysis of
variance showed a significant effect between treatments in final weight, weight gain and survival rate,
with an increase in the values to raise the percentage of S. platensis in diets; however, when compared
to the control diet, values (although lower) near the diet containing 40% were observed, showing a
better amino acid profile in shrimp and providing better productive performance among diets.
Key words: Food, shrimp, growth, yield performance.
INTRODUCTION
Aquiculture has been considered one of the most efficient
paths for a reduction in the deficit between demand and
supply of fish in various regions worldwide. In this
context, shrimp culture stands out as the main segment
due to its accelerated growth rate and high productivity
(MPA, 2010). In Brazil, although recently, this activity is
booming, standing out especially in the North Eastern
region with higher production in the states of Rio Grande
do Norte and Cear芍, obtained by the performance of the
most cultivated peneaeid, the Litopenaeus vannamei
(Natori et al., 2011). However, in cultivation systems,
feeding costs are still the main obstacle among
production costs, representing about 60%, with protein
being the most important and expensive nutrient in
formulated diets (Teixeira et al., 2008).
In this context, many studies have used a great variety
of ingredients previously tested worldwide as protein
source in diets for aquatic species; however, in shrimps,
the proportion has been smaller due to the limited
application of results in the light of the diversity in experimental conditions and lack of nutritional characterization
of ingredients (Glencross, 2006), since the protein source
demands eliminate pollution potential, desirable nutritional
aspects as high protein level, favorable amino acid
profile, high digestibility and acceptable palatability
(Gatlin et al., 2007).
Currently, different microorganisms have been
produced in commercial scale, resulting in an activity of
great interest for the development of different lines within
aquiculture, such as microalgae with great importance in
various trophic chains, especially in the nutrition of other
organisms, treatment of wastewater, production of energy
and in the production of nutritive compounds. Since they
have a very active and low-cost metabolism, they are
able to grow under simple conditions (Derner, 2006),
optimizing their growth according to temperature,
radiation intensity, salinity, agitation, concentration and
nature of nutrients in laboratory conditions (Louren?o,
2006). Among the several microalgae, S. platensis has
*Corresponding
author.
ruthgomesgadelha66@.
E-mail:
Gadelha et al.
274
presented greater nutritional potentials as source of
proteins, vitamins, polyunsaturated fatty acids, besides
having a natural mixture of biopigments with functional
powers (Niu et al., 2007) in the diets of various aquatic
animals, mostly fish, leading to significant effects on
growth, survival rate, immune system and color after
ingestion (Moreira et al., 2010, 2011; Converti et al., 2006).
Based on these data, this study aimed to evaluate the
efficiency of Spirulina platensis at different concentrations
in the diets on the productive performance of L. vannamei
marine shrimp.
MATERIALS AND METHODS
Experimental design and settlement
The experiments were performed at the Research and
Food Processing Center (NUPPA) of the Federal
University of Para赤ba (UFPB), Jo?o Pessoa, Brazil lasting
45 days. 30 L polyethylene boxes were used in open
system with constant aeration from air compressor
attached to a hose and distributed to each tank with
porous stones for oxygen dissipation and artificial lighting,
with a 24-hour photoperiod. The design was entirely
random, consisting of five treatments with three
repetitions. L. vannamei juveniles were acquired from a
cultivation farm located in the city of Livramento, with an
average weight of 1.42㊣0.23g, being acclimatized for two
days, gradually decreasing salinity to 2.5%, being
distributed in their respective treatments with density of
10 shrimps/treatment.
Formulation and elaboration of diets
Five isoproteic diets (35% crude protein) and isoenergetic
(3400 kcal of DE kg-1) were formulated using a specific
software (CRAC version 4.0) with different percentages of
lyophilized S. platensis microalga (SPLF): F (0%), F
(10%), F (20%), F (30%) and F (40%), submitted to the
pelleting process, where dry ingredients were crushed,
weighted, and mixed in an industrial planetary mixer
along with vitamin supplementation (premix) and oil,
adding water at 60∼C until it became consistent
moistened mass. The mixture was introduced in a
manually operated meat grinder in order to form pellets of
2 mm diameter that were further dried in an oven with
forced air circulation, at 80∼C for 24 h and stocked in
paper bags at room temperature.
Feeding
The experimental diets were offered three times a day ad
libitum, and for comparison purposes, commercial diet
(CF) containing 35% crude protein was also used.
Siphoning was performed every two days for the removal
of the remaining diets and feces, followed by the removal
of water samples for the measurement of physicochemical parameters.
Physicochemical analyses of water
The physicochemical analyses of water were performed
throughout the entire cultivation in the Food Science
Laboratory (LACA), with weekly readings of the dissolved
oxygen and temperature by digital oximeter (QUIMISQ758P). pH was measured by a QUIMIS digital pH meter
- both bench type - and elimination of salinity was read
twice a week by a refractometer model BR11 from 0 to
3.5㏑. The quality of water was maintained by daily
renewal of 10% of the volume of each tank, and by
siphoning, and elimination of waste and food debris.
Proximate composition of diets
The proximate composition of diets was determined in
triplicate by analyzing moisture, ash, fat and protein
according to methods described by AOAC (2000).
Amino acids composition of farmed shrimp
The levels of amino acids of the cultivated shrimp were
determined using the methodology of White et al. (1986),
being determined in the sample previously hydrolyzed in
6N redistilled chloric acid, followed by a pre-column
derivatization
of
free
amino
acids
with
phenylisothiocyanate (PITC). The determination of
derivatives of phenylthiocarbamil amino acids (PTC-aa)
was performed in liquid chromatography (VARIAN,
Waters 2690, California, USA).
Growth performance
Biometrics was performed at the beginning and once
every fifteen days for growth performance evaluation,
calculated by:
Survival rate (%) = (final number of shrimp/initial number
of shrimp) ℅ 100; Weight gain (%) = (final average weight
每 initial average weight); SGR (% day - 1) = ln final
weight 每 ln initial weight ℅ 100t; Feed conversion rate =
(food consumption/weight gain).
Statistical analysis
The shrimp growth variables and water quality
parameters were analyzed using analysis of variance
(ANOVA) in order to determine the effect of the levels of
protein substitution and the influence of treatments on
growth, considering the significance level of 5%. In cases
where there was a significant difference, the Tukey＊s test
was applied to compare the average values to the
significance level of 5% using the Statistica R software
version 3.0.
RESULTS AND DISCUSSION
Water quality
The water quality values did not vary significantly
Int. J. Agric. Sci. Res.
275
Table 1. Formulation of the experimental diets used in the experiment.
Ingredients (%)
Lyophilized Spirulina
Fishmeal
Cassava starch
Corn flour
Soybean oil
Corn cobs
Blood meal
Soybean flour
Premix
Salt
F0
0.0
25.0
3.0
25.0
4.0
5.0
15.0
21.5
1.0
0.5
F10
5.2
23.0
2.0
25.0
4.0
4.0
14.0
21.3
1.0
0.5
F20
10.3
20.0
2.0
25.0
4,0
4.0
13.2
20.0
1.0
0.5
F30
15.5
17.0
2.0
25.0
3.5
3.5
12.0
20.0
1.0
0.5
F40
20.7
16.9
2.0
25.0
3.0
3.5
9.4
18.0
1.0
0.5
Chemistry (%)
Crude protein
Lipids
Humidity
Ashes
35.2
6.9
10.4
12.3
35.9
7.1
11.8
11.3
35.8
7.3
11.7
11.5
35.9
6.8
11.3
12.0
35.8
6.7
11.6
11.8
Table 2. Mean ㊣ standard deviation of physicochemical parameters of water in the cultivation of juveniles fed with
different diets.
Diets
F0
F10
F20
F30
F40
*CF
pH
7.75㊣0.04?
7.67㊣0.20?
7.65㊣0.07?
7.73㊣0.08?
7.78㊣0.10?
7.56㊣0.04?
-1
Dissolved Oxygen ((mg.L )
6.48㊣0.19?
6.56㊣0.10?
6.32㊣0.18?
6.28㊣0.26?
6.52㊣0.11?
6.24㊣0.21?
Temperature (∼C)
28.26㊣0.50a
28.00㊣ 0.69?
27.80㊣0.63?
27.32㊣0.41?
28.22㊣0.40?
27.94㊣0.26?
*Commercial diet. Same letters in the same column are not statistically different (P>0.05).
throughout the experimental period during cultivation
(Table 2) of L. vannamei, with temperature values ranging
from 28 to 30∼C, pH between 6 and 9, and dissolved
oxygen above 4 mg/L according to Boyd (2002). This
same author recommends 2.5% salinity as pattern for the
achievement of greater shrimp growth, also emphasizing
that this species is adapted to lower salinity levels.
Chemical composition of diets
The chemical composition of the experimental diets is
shown in Table 1, with protein values around 35%. This
percentage was considered ideal for the development in
the juvenile phase of L. vannamei according to RivasVegas et al. (2006) and Holme et al. (2009). For humidity,
values below 13% were observed to be commonly found
in commercial diets, and were considered appropriate for
the formulation of diets for aquatic animals avoiding
excessive drying and decreased protein quality of pellets,
according to Cuzon and Guillaume (1997).
The lipid values ranged from 6.7 to 7.1%, which are in
agreement with values required by this species (6 to
7.5%). According to Gonz芍lez-F谷lix et al. (2002), these
values are considered ideal to avoid a decrease in growth
and increase in mortality rate of shrimps; in relation to the
ash values, diets showed values similar to those found in
commercial diets, with maximum of 13%, observed by
Melo (2003), when commercial diets for shrimps were
analyzed.
Amino acid profile
In the muscle tissue composition of L. vannamei (Table 3)
marine shrimp, higher concentrations of arginine, lysine
and methionine were observed in animals fed with diets
supplemented with 10 and 40% of S. platensis. However,
it was found that the methionine content in animals fed
with diet supplemented with 10% was lower when
compared to the muscle demand values for this shrimp
species fed with commercial diet. Whereas the imbalance
Gadelha et al.
276
Table 3. Amino acid profile in the muscle of the juvenile shrimp fed with experimental diets using SPLF.
AARP (%)
Arginine
Phenylalanie
Histidine
Isoleucine
Leucine
Lysine
Methionine
Tyrosine
Threonine
Valine
*Shrimp
7.5
3.6
1.9
3.6
6.5
6.4
2.6
5.4
3.4
3.8
F0
4.9
2.7
1.4
2.5
3.7
5.3
1.6
2.0
1.8
2.7
F10
8.8
4.0
2.1
3.9
5.9
8.6
2.2
3.3
3.0
3.9
F20
2.3
1.1
1.5
1.1
1.8
0.4
0.4
0.8
0.3
1.6
Treatment
F30
0.06
2.0
1.2
1.8
2.9
1.1
1.2
1.6
1.7
2.0
F40
8.3
5.5
5.6
4.4
6.1
8.5
2.8
4.3
4.0
5.4
FC
9.6
6.0
3.0
5.6
6.2
12.8
3.5
4.2
4.0
5.7
*Shrimp muscle requirement (Cuzon et al., 2004).
Table 4. Production parameters of Litopenaeus vannamei juveniles fed with experimental diets.
Supplementation
(%)
Final weight
(g)
Final length
(cm)
0
10
20
30
40
*CF
3.59 ㊣ 0.25 d
3.72 ㊣ 0.19 cd
3.93 ㊣ 0.33 cd
4.07 ㊣ 0.41bc
4.43 ㊣ 0.37 b
4.97 ㊣ 0.52 a
8.49㊣0.51b
8.60㊣0.51b
8.88㊣0.35ab
8.91㊣0.62ab
8.97㊣0.37ab
9.25㊣0.25a
Weightgain
(g)
2.17 ㊣ 0.35 d
2.29 ㊣ 0.27 cd
2.49 ㊣ 0.37 cd
2.65 ㊣ 0.50 bc
3.01 ㊣ 0.43b
3.53 ㊣ 0.53 a
FCR
SGR (day-1)
1.3a
1.1a
1.3a
1.2a
1.2a
1.7b
2.04 ㊣ 0.37 c
2.13 ㊣ 0.24 c
2.22 ㊣ 0.33 bc
2.33 ㊣ 0.48 bc
2.51 ㊣ 0.43 ab
2.55 ㊣ 0.41 a
Survival rate
(%)
85d
90c
85d
97?
96ab
95b
Same letters in the same column are not statistically different. (P>0.05). *Commercial diet. FCR= Feed Conversion rate.
in amino acids, even if it is represented by a single
essential amino acid, has an immediate effect on meeting
the protein needs. As regards the energy balance (Fox et
al., 2004), it was observed that the diet containing 40% S.
platensis provided the best concentrations of amino acids
for growth performance in animals.
According to Holme et al. (2009), diets containing
amino acids in proportions similar to those in the shrimp
muscles provide the best growth and survival rates during
commercial farming, and the diet quality was not
necessarily related to the total amount of proteins, but a
well-balanced supplementation of amino acids can be
found in S. platensis present in its composition, with a
complete protein containing all the essential and
nonessential amino acids (Di Lifetec Co LTD, 2009).
Production performance indicators
At the end of the 45 days of experiment, significant
difference was observed due to the fact that as the final
weight and weight gain in the shrimp increased, the
percentage of S. platensis in the diet increased, indicating
a better outcome in these parameters with percentages of
30 and 40%. However, when compared to the commercial diet, lower results were observed, but with values
close to those obtained with 40% supplementation. The
literature has shown better results for growth with
supplementation of S. platensis up to 25% in the diet for
several fish species, but similar or better results
compared to this study can be found if the percentage of
this microalga is increased in diets (Table 4).
According to Abdel-Tawwab and Ahmad (2009), these
positive effects on growth are attributed to nutrient
digestibility and high nutritional content present in S.
platensis. The mean survival rate of shrimp showed
significant difference, and the results were not influenced
by the percentage of S. platensis in diets, with high
values, which show the positive effects of this microalga
to promote better cultivation conditions. Jaime-Ceballos
et al. (2007) reported that a 25% increase of these
microalgae in the diet of post-larvae Litopenaeus schmitti
showed higher growth and survival rates. Ghaeni et al.
(2011) used S. platensis in the diet of tiger shrimps
Penaeus semisulcatus and obtained better weight gain
results. Most researchers have used microalgae to feed
fish including tilapia, however, Gomes et al. (2012) and
Ungsethaphand et al. (2010) reported greater weight gain
efficiency and survival rates.
Higher feed conversion rate was observed in animals
fed with commercial diet; however, diets containing S.
Int. J. Agric. Sci. Res.
platensis showed satisfactory results according to Boyd
(1997), indicating good feed conversion rates from 0.9 to
1.5, which van vary depending on the storage density of
diets. The same author reported that shrimp cultivation
farms obtain feed conversion rate from 2.0 to 2.4. SilvaNeto (2010) evaluated S. platensis as attractive in diets
for L. vannamei and showed better feed conversion rates
in animals, concluding that this micro alga has the ability
to stimulate food intake in animals even at low inclusion
levels in the diet.
Conclusion
Based on the results obtained in the present study, it
could be concluded that the use of S. platensis in the
cultivation of L. vannamei marine shrimps showed
positive effects on water quality and a food supplement
by providing greater increase in growth, weight and
length, with high survival rate in all treatments. However,
the 40% level showed a better amino acid profile, with
more significant results in the productive performance of
shrimps. Further studies should be carried out using S.
platensis as a food supplement with higher percentage
compared to this research.
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