EFFECT OF USING DIFFERENT LEVELS OF Spirulina platensis ON GROWTH PERFORMANCE OF MARINE SHRIMP Litopenaeus vannamei

Full text

Egyptian J. Nutrition and Feeds (2021), 24(3): 431-437
Issued by The Egyptian Society of Nutrition and Feeds
EFFECT OF USING DIFFERENT LEVELS OF Spirulina platensis ON
GROWTH PERFORMANCE OFMARINE SHRIMP Litopenaeus vannamei
Shimmaa A.H. Zidan1 , A.E. Eid1, Mervat A. M. Ali1 and Zaki Z. Sharawy2
1Faculty of Agriculture, Suez Canal University, Egypt.
2Invertebrate Laboratory, Aquaculture Division, National Institute of Oceanography and Fisheries
(NIOF), Egypt.
(Received

2/8/202

, accepted 19/9/202

)
SUMMARY
his study was designed to evaluate the effect of replacement of fish meal with a marine
microalgae species spirulina (Spirulina platensis) on Pacific white
shrimp Litopenaeus vannamei post larvae for 90 days. Spirulina was used with four levels
(5, 10, 15 and 20). Experiment was carried out in 15 tanks with water volume 150 L Each treatment
had 3 replicates, each tank contained 50 Post-larvae of 0.02 g body weight. Shrimp were fed the
experimental diets containing about 40% protein twice daily at 14% from body weight (initial weight)
and readjusted gradually to 5% at the end of the experiment. Growth performance of shrimp was
recorded biweekly. It was found that the best treatment of growth performance,feed utilization and
economic evaluation under these experimental conditions was the treatment in which fish meal was
replaced by 10% spirulina.
Keywords: L. vannamei, Spirulina platensis, growth performance, feed utilization and economic
evaluation.
INTRODUCTION
Pacific white shrimp, Litopenaeus vannamei, is an economically important farm-raised shrimp
because of its great economic value, rapid growth rate and tolerance of a big range of temperatures
and salinities (Huang et al., 2015). In 2010, Litopenaeus vannamei accounted for 71.8% of world
production of all farmed marine shrimp species (FAO, 2012). To increase the growing market for
cultured shrimp, the demand for improved feeds has created a need for high-quality protein sources
(Tacon & Forster, 2000). Fish meal is considered as primary ingredient in marine shrimp diets
because of its balanced amount of essential amino acids, fatty acids, minerals, vitamins and
palatability (Suárez et al., 2009). Commercial shrimp feed formulations generally include between
25% and 50% fish meal, which considered the primary and most expensive ingredient (Gonzalez-
Rodriguez & Abdo de la Parra, 2004). The increasing demand for fish meal and overexploitation of
fish stocks, has spurred a search for sustainable and cheaper protein ingredients to reduce or eliminate
the use of fish meal in aquaculture diets (Kiron et al., 2012).
Microalgae are known as a potential source of food and energy due to their photosynthetic
efficiency and high nutritional value. The high protein contents of different microalgae species are
considered as promising substitutes for fish meal protein or as a valuable additive in aqua feeds (Ju et
al., 2012). In addition, microalgae are the rich source of proteins, vitamins, fatty acids and minerals
(Radhakrishnan et al., 2014), they may possibly be an ideal alternative ingredient for sustainable
aquaculture feeds. In terms of the amino acid profile, almost all microalgae compare positivly with
that of other food proteins (Becker, 2007).
Among many types of microalgae that considered for feed complement in fish and shrimp culture
until now, Spirulina platensis is known as one of the richest sources of protein, vitamins, minerals,
essential amino acids, fatty acids and antioxidant pigments, like carotenoids (Radhakrishnan et al.,
2014). Spirulina protein has a balanced composition of amino acids and concentrations of methionine,
tryptophan and other amino acids similarity to these of casein. It can be used as partial
T

Zidan et al.
432
supplementation or complete replacement for protein in aqua-feeds and is a cheaper feed ingredient
than another animal origin (Habib et al., 2008).
The aim of the present study was to evaluate the effect of replacement of fish meal with a
marine microalgae species S. platensis on growth performance, feed utilization and economical
evaluation, in Pacific white shrimp L. vannamei post larvae.
MATERIALS AND METHODS
The white leg shrimp L. vannamei post-larvae were obtained from a commercial shrimp hatchery
(Berket Ghalioun, Kafr Al-Sheikh, Egypt). Shrimps were transported in double –layered polythene
bags that were oxygenated. When the shrimp arrived at the laboratory, they were moved into the
acclimation tanks filled with seawater (salinity, 32ppt). Before start the experiment, shrimps were
acclimated to laboratory condition for two weeks and were fed twice daily with a commercial diet
(Skretting: 38% crude protein, 8% crude fat, and 5.9 crude fiber with 3980 Kcal of energy).
The experiment was carried out in 15 tanks with water volume 150 L. Each treatment had 3
replicates, each tank contained 50 Post-larvae of 0.02 g body weight. Tanks were filled with seawater
after filtered by plankton net (50µm) to prevent the entry of un wanted materials and suspended
particles into the tanks and was diluted with fresh water to make a salinity (32 ppt). All tanks were
supplied with 3 air stone-hoses type of diffuser system which is fitted to air-blower (220 w). Aeration
was provided 24 hours throughout the experiment. All tanks were always covered with plastic sheet to
reduce escapes of shrimp. water was exchanged once a week.
After two weeks, all tanks were stocked with shrimp post-larvae. Shrimp were fed the
experimental diets (as shown in Table 1) twice daily at 14% from body weight (initial weight) and
decreasing gradually to 5% at the end of the experiment. Each treatment's daily feeding ration was
calculated and adjusted by estimating the biweekly sampled mean biomass.
Table (1): Composition and proximate analysis of the experimental diets (% on DM basis).
Ingredient
Treatments
Control
T1 (5%) 
T2 (10%)
T3 (15%)
T4 (20%)
fish meal (70%) protein

34.00
30.00
27.00
23.00
20.00
Soybean meal (44%) protein

34.00
34.00
34.00
34.00
34.00
Commercial Spirulina 
00.00
4.00
7.00
11.00
14.00
Yellow corn
23.00
23.00
23.00
23.00
23.00
Sun flour oil
6.00
6.00
6.00
6.00
6.00
Mineral mixture1
2.00
2.00
2.00
2.00
2.00
Vitamin mixture2
1.00
1.00
1.00
1.00
1.00
Total
100
100
100
100
100
Proximate analysis
Protein
41.39
40.00
39.55
39.35
39.89
Lipids
10.32
10.31
10.32
10.36
10.35
Ash
7.35
7.31
7.32
7.34
7.36
Fibers
3.16
3.24
3.36
3.44
3.54
Mositure
23.71
23.69
23.43
23.81
23.80
NFE3
37.78
39.14
39.45
39.51
38.86
Gross energy
(Kcal/100g) 4
486.65
484.29
483.12
482.62
482.90
1Each Kg mineral mixture premix contained Mn, 22 g; Zn, 22 g; Fe, 12 g; Cu, 4 g; I, 0.4 g, Selenium, 0.4 g and
Co, 4.8 mg.
2Each Kg vitamin contained Vitamin A, 4.8 million IU, D3, 0.8 million IU; E, 4 g; K, 0.8 g; B1, 0.4 g; Riboflavin,
1.6 g; B6, 0.6 g, B12, 4 mg; Pantothenic acid, 4 g; Nicotinic acid, 8 g; Folic acid, 0.4 g Biotin,20 mg

3Nitrogen Free Extract = 100 – (%Protein + %Fat + %Fiber + %Ash).
4Gross Energy based on protein (5.65 Kcal/g), fat (9.45 Kcal/g) and carbohydrate (4.11Kcal/g). According to (NRC, 2011)
The experimental diets were prepared by weighing of each component individually and mixing the mineral,
vitamins and additives with corn. Then, this mixture was added to the components together with oil. Water was

Egyptian J. Nutrition and Feeds (2021)
433
added to the mixture until became suitable for making granules. This mixture was passed through CBM granule
machine with 2mm diameter. The pellets were dried at room temperature and kept frozen until experimental
start.
Spirulina (Arthrospira platensis) used in present study was obtained from local market (Alhlw, Co. for
biological production-Zagazik, Egypt). The chemical analysis of Spirulina (Arthrospira platensis) was shown in
Table (2).
Table (2): The Chemical analysis of Spirulina (Arthrospira platensis).
Growth performance parameters:
Shrimp weight (g) was measured at the initial of the experiment and biweekly by collected
randomize number of shrimp from each tank and weighted in an analytical digital balance and then
returned back to their tanks during the experiment. Shrimp Weight gain (WG), Specific growth rate
(SGR) and Survival rate% (SR) were calculated according to the following equations:
 Weight gain (WG) = Final body weight (g) - Initial body weight (g).
 Specific growth rate % (SGR) = [(ln FBW - ln IBW) /day of experiment] ×100
 Survival rate

(SR) = (Final number of shrimp / Initial number of shrimp) ×100.
Feed utilization parameters:
Feed utilization parameters were calculated according to the following equations:
 Feed Conversion Ratio (FCR)= Total feed consumption/ weight gain.
 Feed Efficiency (FE) %= (Final weight – initial weight) / feed consumed ×100.
 Protein Efficiency Ratio (PER)= body weight gain (g)/ protein intake (g)
Statistical analysis:
All data of variables measured were analyzed by two-way ANOVA. The ANOVA was performed
by using the SAS v9.0.0 (SAS, 2004) program. The ANOVA was followed by Duncan's test (Duncan,
1955) at P<0.05 level of significance.
Economical evaluation:
The cost of feed to raise unit biomass of shrimp was estimated by a simple economic analysis. The
estimation was based on local retail sale market price of all the dietary ingredients at the time of the
study.
 Cost /kg diet (LE) = Cost per Kg diet L.E.
 Consumed feed to produce 1kg shrimp (kg) = Feed intake per shrimp per period/ final weight per
shrimp Kg/Kg.
 Feed cost per kg fresh shrimp (LE) = Step 1× step 2.
 Relative % of feed cost/ kg shrimp = Respective figures for step 3/ highest figure in this step.
 Feed cost 1Kg gain (LE) = Feed intake per Kg gain × step 1.
 Relative % of feed cost of Kg gain = Respective figures for step 5/ highest figure in this step.
Component
% (Mean ± SD)
protein
57.79 ± 1.53
Carbohydrates
14.60 ± 0.60
Ash
12.05 ± 0.31
Lipids
9.33 ± 0.16
Moisture
6.98 ± 0.05
Fibers
6.23 ± 0.25

Zidan et al.
434
RESULTS AND DISCUSSION
Growth performance and feed utilization:
Table (3) presents the effect of different dietary replacement of spirulina platensis protein levels
on growth performance and feed utilization in experimental tanks of L. vannamei . After the 90 days
feeding trial, mean weight gain of the shrimp fed with diet T1 and T2 were differed significantly
(P<0.05) than control diets. The spirulina diets with the replacement of fish meal at 5 and 10%
showed higher weight gain; 8.55±0.08g for T1 and 8.63±0.72g for T2. The 10% treatment (T2) had
significantly (P<0.05) the best feed efficiency (FE) and protein efficiency ratio (PER) compared with
the rest of experimental groups.
Table (3): Effect of using spirulina platensis protein levels on growth performance and feed
utilization (Mean±SD) in experimental tanks of L. vannamei for 90 days.
Data are presented as means ±SD. Values in the same row with different superscript letters are significantly
different (P< 0.05).
In this study, best growth rates and most efficient FCR were achieved at T2. Growth enhancement
effect of spirulina is because of its role in nutrient digestibility and its high contents of several
nutrients, like vitamins and minerals (Abdel-Tawwab and Ahmad, 2009). On the other hand, negative
effects of high dietary inclusion levels of spirulina on fish growth can be resulted from reduced
phosphorous availability and decreased feed palatability (Olvera-Novoa et al., 1998). The variations
in spirulina effects on fish growth performance are attributed to different nutrient content of spirulina
species used in the studies (Nandeesha et al., 1998). Nandeesha et al. (2001) found that fish meal
(FM) can be completely replaced with spirulina in diets for rohu carp (Labeo rohita) and even
significantly higher growth can be obtained compared to the use of FM as the sole protein source,
whereas no significant effect was observed on growth performance of catla (Catla catla) by the same
spirulina supplemented diets. Such as differences in growth response of L. rohita and C. catla to
dietary spirulina clearly show that the growth response of fishes to spirulina is likely to be species-
specific. The other significant factor that affects the results of spirulina administration is the
composition of experimental diets in which spirulina is combined (Takeuchi et al., 2002).
James et al. (2006) observed that 8% of spirulina diet showed higher food consuming rate in
Xiphophorus helleri. Rhabdosargus sarba fingerlings fed with 32% spirulina feed had higher feed
intake (El-Sayed, 1994). The study is in agreement with the report by Nakagawa and Gomez-Diaz
(1995) who found that the diet with 5 – 20% supplementation of spirulina meal improved the
Macrobrachium rosenbergii survival and growth. The results showed improvement of fish growth by
replacing 5% -10 % FM protein with spirulina, while higher substitution levels could not provide
further enhancement. Tongsiri et al. (2010) observed that replacement of 5% FM with spirulina
showed the best growth performance of P. gigas, but higher replacement levels lowered the fish
weight gain. Olvera-Novoa et al. (1998) reported that spirulina can replace up to 20% of FM protein
Parameter
Treatment
Control
T1
5%
T2
10%
T3
15%
T4
20%
IBW (g)
0.02
0.02
0.02
0.02
0.02
FBW (g)
7.86±0.71c
8.57±0.08a
8.65±0.92a
8.33±1.7 b
7.32±1.58c
WG (g)
7.84±0.71c
8.55±0.08a
8.63±0.72a
8.31±1.6 b
7.31±1.18c
SGR (%/day)
6.93±0.67 b
7.02±0.69a
7.03±0.67a
6.99±0.69b
6.85±0.41b
FI (g feed/shrimp)
10.42±0.72c
11.83±0.54a
10.18±0.24c
11.73±0.93b
10.16±0.24c
FCR
1.33±0.19b
1.38±0.07a
1.18±0.09c
1.41±0.23a
1.39±0.12a
PER
1.82±0.21b
1.81±0.35b
2.14±0.23a
1.80±0.24b
1.80±0.12b
FE (%)
75.27±2.3b
72.26±2.51b
84.81±1.89a
70.87±1.35c
71.92±1.23c
SR (%)
92.67±3.56b
94.67±4.65a
94.67±6.42a
92.00±3.49b
90.67±3.7c

Egyptian J. Nutrition and Feeds (2021)
435
in diets for O. mossambicus, but reduced growth and feed utilization were showed at higher
replacement levels. Güroy et al. (2012) also observed better growth in yellow tail cichlids, with an
increase in weight of between 12% and 17% following the inclusion of 2.5% and 10% Spirulina,
respectively. In an even better scenario, Adel et al. (2016) observed that sturgeon juveniles fed with
10% Spirulina showed a growth increase of 57% and an improvement in the FCR of 21%. Another
important study by Kohal et al. (2018) reported that the red cherry shrimp showed a dramatic
improvement in growth from 70.8 mg to 114.6 mg following the addition of 10% Spirulina to the diet,
which represents a gain of 63%, an improvement in the FCR of 15%, and a survival increase from
25.7% on the control diet to 81.3% on the 10% Spirulina diet. Partial substitute of fish meal with S.
platensis has showed promising growth in juvenile L. vannamei (Hanel et al., 2007). The admissible
level of spirulina meal as the dietary inclusion to replace the fishmeal was about 25% with no harmful
effect on the growth of shrimp L. vannamei, and up to 50% of replacement could not affects the feed
intake (Sá and Nunes, 2011). Many such examples explain that if Spirulina is used at low levels in fish
diets could get not only health benefits but also economic benefits by improving the FCR.
Economical Evaluation:
Calculations of economic efficiency of the tested diets based on the cost of feed, costs of one Kg
gain in weight and its ratio with the control group are shown in Table (4). T2 have the lowest feed cost
per kg fresh shrimp (18.59 LE), relative % of feed cost / kg shrimp (85%), feed cost /1Kg gain (18.59
LE) and relative % of feed cost of Kg gain (85%).
Table (4): Economic analysis of Litopenaeus vannamei supplemented with different levels of
Spirulina for 90 days.
Item
Treatment
Control
T1
5%
T2
10%
T3
15%
T4
20%
Cost /kg diet (LE)
16.37
15.93
15.75
15.31
14.93
Consumed feed to produce 1kg shrimp (kg)
1.33
1.38
1.18
1.41
1.39
Feed cost per kg fresh shrimp (LE)
21.77
21.98
18.59
21.59
20.75
Relative % of feed cost/kg shrimp
99
100
85
98
94
Consumed feed to produce 1Kg gain (Kg)
1.33
1.38
1.18
1.41
1.39
Feed cost /1Kg gain (LE)
21.77
21.98
18.59
21.59
20.75
Relative % of feed cost of Kg gain
99
100
85
98
94
CONCLUSION
It can be concluded that the diet in which fish meal was replaced with 10% spirulina was the best
in terms of growth performance, feed utilization and economic evaluation under these experimental
conditions.
REFERENCES
Abdel-Tawwab, M. and H. Ahmad (2009). Live Spirulina (Arthrospira platensis) as a growth and
immunity promoter for Nile tilapia, Oreochromis niloticus (L.), challenged with pathogenic
Aeromonas hydrophila. Aquaculture Res., 40(9):1037-1046.
Adel, M., S. Yeganeh, M. Dadar, M. Sakai and M. A. Dawood (2016). Effects of dietary Spirulina
platensis on growth performance, humoral and mucosal immune responses and disease resistance
in juvenile great sturgeon (Huso huso Linnaeus, 1754). Fish & Shellfish Immunology, 56: 436–
444.
Becker, E. W. (2007). Micro-algae as a source of protein. Biotechnology Advances, 25, 207– 210.
Duncan, D. B. (1955). Multiple range and multiple F test. Biomertics, 11(1):1-42.

Zidan et al.
436
El-Sayed, A. F. M .(1994). Evaluation of soybean meal, Spirulina meal and chicken offal meal as
protein sources for silver sea bream (Rhabdosargus sarba) fingerlings. Aquaculture, 127: 169–
176.
FAO, (2012). FAO Statistical Yearbook: Fishery and Aquaculture Statistics. The organization of Food
and Agriculture of the United Nations, Rome.
Gonzalez-Rodriguez, B. and I. Abdo de la Parra (2004). Replacement of fish meal with co-extruded
wet tuna viscera and corn meal in diets for white shrimp (Litopenaeus vannamei,
Boone). Aquaculture Research, 35, 1153– 1157.
Güroy, B., I. Sahin, S. Mantoglu and S. Kayalı (2012). Spirulina as a natural carotenoid source on
growth, pigmentation and reproductive performance of yellow tail cichlid Pseudotropheus
acei.Aquaculture International 20: 869–878.
Habib, M.A.B., M. T.C. Parvin, K. Huntington and M.R. Hasan (2008). A review on culture,
production and use of Spirulina as food for humans and feeds for domestic animals and fish. FAO
Fisheries and Aquaculture circular no 1034. Rome, FAO: 41.
Hanel, R., D. Broekman, S. de Graaf and D. Schnack (2007). Partial Replacement of Fishmeal by
Lyophylized Powder of the Microalgae Spirulina platensis in Pacific White Shrimp Diets. The
Open Marine Biology Journal, 2007, 1, 1-5.
Huang, X. L., M. H. Xia, H. L. Wang, M. Jin, T. Wang and Q. C. Zhou (2015). Dietary thiamin could
improve growth performance, feed utilization and non-specific immune response for juvenile
pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition, 21, 364– 372.
James, R., K. Sampath, R. Thagarathinam and I. Vasudeven (2006). Effect of dietary Spirulina level
on growth, fertility, coloration and leucocyte count in Red Swordtail, Xiphophorushelleri. Isr. J.
Aqua - Bamidgeh. 58 (2): 97-104.
Ju, Z. Y., D. F. Deng and W. Dominy (2012). A defatted microalgae (Haematococcus pluvialis) meal
as a protein ingredient to partially replace fishmeal in diets of Pacific white shrimp (Litopenaeus
vannamei, Boone, 1931). Aquaculture, 354–355: 50– 55.
Kiron, V., W. Phromkunthong, M. Huntley, G. Archibald and G.D. Scheemaker (2012). Marine
microalgae from biorefinery as a potential feed protein source for Atlantic salmon, common carp
and white leg shrimp. Aquaculture Nutrition, 18: 521– 531.
Kohal, M.N., A.E. Fereidouni, F. Firouzbakhsh and I. Hayati (2018). Effects of dietary incorporation
of Arthrospira (Spirulina) platensis meal on growth, survival, body composition, andreproductive
performance of red cherry shrimp Neocaridina davidi (Crustacea, Atyidae) over successive
spawnings. Journal of applied phycology, 30:1–13.
Nakagawa, H. and G. Gomez-Diaz (1995). Usefulness of Spirulina sp. meal as feed additive for giant
freshwater prawn, Macrobrachium rosenbergii. Suisanzoshoku, 43:521-526.
Nandeesha, M.C., B. Gangadhara, J.K. Manissery and L.V. Venkataraman (2001). Growth
performance of two Indian major carps, catla (Catlacatla) and rohu (Labeorohita) fed diets
containing different levels of Spirulina platensis. Bioresour Technol., 80(2):117-20.
Nandeesha, M. C., B. Gangadhara, T. J. Varghese and P. Keshavanath (1998). Effect of feeding
Spirulina platensis on the growth, proximate composition and organoleptic quality of common
carp, Cyprinuscarpio. Aqua cult. Res., 29: 305-312.
NRC (2011). National Research Council,Nutrient Requirement of fish National Academic Prees,
washington, DC.
Olvera-Novoa, M. A., L. J. Dominguez-Cen, L. Olivera-Castillo and C.A. Martmez-Palacios (1998).
Effect of the use of the microalga Spirulina maxima as fish meal replacement in diets for tilapia,
Oreochromismossambicus (Peters), fry. Aquaculture Research, 29:709-715.
Radhakrishnan, S., P. Saravana Bhavan, C. Seenivasan, R. Shanthi and T. Muralisankar (2014).
Replacement of fishmeal with Spirulina platensis, Chlorella vulgaris and Azolla pinnata on non-
enzymatic and enzymatic antioxidant activities of Macrobrachium rosenbergii. The Journal of
Basic & Applied Zoology, 67: 25– 33.
Sá, M. V. C. and A.J. P. Nunes (2011). Spirulina meal, feeding attractant spare fishmeal in white
shrimp diets. Global Aquacul. Advocate, May/June, 34-35.

Egyptian J. Nutrition and Feeds (2021)
437
SAS (2004). Statistical Analysis System Institute. User׳s Guide: statistics. SAS Institute Inc., Cary.
Suárez, J.A., G. Gaxiola, R. Mendoza, S.Cadavid, G. Garcia, G.Alanis and G.
Cuzon (2009). Substitution of fish meal with plant protein sources and energy budget for white
shrimp Litopenaeus vannamei (Boone, 1931). Aquaculture, 289: 118– 123.
Tacon, A.G.J. and I.P. Forster (2000). Trends and challenges to aquaculture and aquafeed
development in the new millennium. In: L.E. Cruz-Suarez, D. Ricque-Marie, M. Tapia-Salazar,
M.A. Olvera-Novoa, and R. Civera- Cerecedo (editors). Avances en Nutrición Acuicola V.
Memorias del V Simposium Internacional de Nutrición Acuicola, Mérida, Yucatan, Mexico. pp. 1
12.
Takeuchi, T., J. Lu, G. Yoshizaki and Y. Satoh (2002). Effect on the growth and body composition of
juvenile tilapia Oreochromis niloticus fed raw Spirulina. Fish. Sci., 68:34- 40.
Tongsiri, K., Mang-Amphan and P. Yuwadee (2010). Effect of Replacing Fishmeal with Spirulina on
Growth, Carcass Composition and pigment of the Mekong Giant Catfish. Asian Journal of
Agricultural Sciences, 2(3): 106-110.





















09
          
    


 


