Live Feed for sustainable Aquaculture

Micro-algae are an essential food source in the rearing of all stages of shrimp, Rotifers and
zooplankton.
They can be used as essential live feeds and supplements in the culture of larval and juvenile rotifers
and Shrimp.
Non-toxic marine microalgae, including the stramenopiles Isochrysis sp., Pavlova sp. and
Nannochloropsis sp. as well as various diatoms, represent the primary food source at some stages in
the life cycle of most cultivated marine animals
[Graham JE, Wilcox LW, Graham LE: Algae. City: Pearson Education; 2009.].
Fishmeal, groundnut meal and soybean meal can be partially replaced by spirulina in the preparation
of diets of fish, poultry, cattle and domestic animals (Venkataraman, Somasekaran and Becker, 1994;
ElSayed, 1994; Britz, 1996).
The total global production of farmed shrimp reached more than 1.6 million tonnes in 2003,
representing a value of nearly US$9,000 million
(http://fisherymanagement.wikia.com/wiki/Shrimp_farm).
Shrimp farming production reached 737,200 tonnes in 1998, an increase of 12% from 1997. This
increased production was mainly reported in subtropical regions of America (28%, 457 hatcheries)
and SE Asia (72%, 3,718 hatcheries)
[Hemaiswarya S, Raja R, Kumar RR, Ganesan V, Anbazhagan C: Microalgae: a sustainable feed
source for aquaculture. World J Microb Biot 2011, 27: 17371746. 10.1007/s11274-010-0632-z].
According to Shrimp Production Review by Food and Resource Economics Department, University of
Florida USA, the shrimp farming was estimated to be about 4 million metric tonnes in 2013, with
10.3% projected annual growth rate (mainly attributed to SE Asian countries and China)
(http://www.gaalliance.org/update/GOAL11/DiegoValderrama.pdf).
However, disease outbreaks in recent years have affected farmed Atlantic salmon in Chile, oysters in
Europe, and marine shrimp farming in several countries in Asia, South America and Africa, resulting
in partial or sometimes total loss of production
(http://www.fao.org/docrep/016/i2727e/i2727e.pdf).
In 2010, aquaculture in China suffered production losses of 1.7 million tons caused by natural
disasters, diseases and pollution. Disease outbreaks virtually wiped out marine shrimp farming
production in Mozambique in 2011 (http://www.fao.org/docrep/016/i2727e/i2727e.pdf).

Microalgae species Hypnea cervicornis and Cryptonemia crenulata particularly rich in protein were
tested in shrimp diets. Amount of algae in fish meal resulted in significant increase in shrimp growth
rates [Harun R, Singh M, Forde GM, Danquah MK: Bioprocess engineering of microalgae to produce
a variety of consumer products. Renew Sust Energ Rev 2010, 14: 10371047.
10.1016/j.rser.2009.11.004].
Spirulina was studied as a feed supplement for the giant freshwater prawn (Macrobrachium
rosenbergii), and found to significantly improve growth, survival, and feed utilization. The
supplementation range was 520 percent and results were similar at any of the ranges added to the
feed
(Nakagawa and Gomez-Diaz, 1975).
Centrifugation of algae into a paste form and subsequent refrigeration is cumbersome and dearer.
Limited shelf-life and the high prices of such presently available algal pastes are of not much use to
farmer.
Outdoor pond production on a large scale of Spirulina and Dunaliella salina are used as feed but
these are also expensive.
Efforts are put to grow micro-algae under heterotrophic growth conditions in dark, by using organic
carbon as an energy source.
However, Species like Chaetoceros, Isochrysis, Skeletonema, Thalassiosira, Monochrysis are
incapable of growing in the dark.

Common algae used as feed to Brooder stock, Rotifer, Shrimp

larvae, growing shrimp.
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Chaetoceros calcitrans
Chaetoceros gracilis
Chaetoceros muelleri
Chaetoceros simplex
Chlamydomonas sp.
Chlorella sp.
Chroomonas salina
Cyclotella sp.
Dicruteria sp.
Dunaliella perva
Dunaliella tertiolecta
Isochrysis galbana
Isochrysis sp., clone T-Iso
Nannochloropsis sp.
Pavlova lutheri
Pavlova salina
Phaeodactylum tricornutum
Skeletonema costatum
Tetraselmis chui
Tetraselmis levis
Tetraselmis suecica
Thalassiosira pseudonana, clone 3H
Thalassiosira Weissfloggii

Advantages of Algal Feed

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Algae will degrade the pollutants and toxins

During Day time Algae will improve Dissolved Oxygen.
Algae is the natural feed for aquatic animals
Algae, having better absorption and better nutrient profile, reduces the need and cost
of Artificial feed.
5. Single source of balanced feed
6. Totally organic and Natural
7. Very rare and valuable components like Astaxanthin, Lutein, Beta-carotene,
Chlorophyll, Phycobiliproteins, Polyunsaturated fatty acids (PUFAs), Beta-1,3-glucan,
sulfolipids , (6-hydroxy-7-O-methylscytophycin B, aurilides B and C etc., are only
available from Algae.

(https://www.researchgate.net/publication/248340580_Water_quality_control_using_Spiru
lina_platensis_in_shrimp_culture_tanks)

Both microalgae and macroalgae play a crucial role in present shrimp culture management
strategies. The recovery and subsequent administration of algal compounds such as
polysaccharides, fatty acids as well as secondary metabolites to shrimp have resulted in
enhanced immune parameters and increased resistance to bacterial and viral pathogens.
Some of these compounds have been known to exhibit either antiviral or antibacterial
activities. This mode of action against pathogens has also been attributed along with
microalgal aggregates and finfish for the prevention and control of vibriosis in the
greenwater culture system. The significant microalgal assemblage that characterizes this
culture system also contributed in maintaining shrimp immunocompetence through
immediate intake of nutrients and through maintenance of good pond water quality.
Overall, these various applications of algae provide a cheaper and environment friendly
solution for the prevention and control of diseases in shrimp aquaculture.

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Isochrysis is a small golden/brown flagellate. It contains higher DHA. Zooplankton like Rotifer,
Artemia love it.
Nannochloropsis is a small green alga. Zooplankton like Rotifer, Artemia love it.
Pavlova is a small golden/brown flagellate and is similar to Isochrysis. It also possesses very high DHA
profile. Useful for enriching rotifers and other zooplankton.
Tetraselmis is a large green flagellate and contains a very high lipid level. Useful as feed for larval
shrimp.
Thalassiosira weissflogii is a large diatom that is used in the shrimp.
Chlorella possesses better absorption. H. Nakamura (1961) recommended Chlorella up to 5 percent
in the daily diet.

COMPARISION OF ALGAE
PER, NPU, and BV Values of Different Micro-algae.
PER

NPU

BV

Spirulina maxim*

2.30

45.6 - 49.8

60 - 65

Spirulina platensis

2.07

52 7

68

Scenedesmus acutus**

1.27

52

72.1

Chlorella ellipsoidea

0.94

* Clement and Van Landeghem (3)

** Becker et al. (4)

CHEMICAL COMPOSITION OF SPIRULINA

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Essential amino acids

ISOLEUCINE (4.130/o): Required for optimal growth, intelligence development and nitrogen equilibrium
in the body Used to synthesize other non-essential amino acids.
LEUCINE (5.8001o): Stimulator of brain function, increases muscular energy levels.
LYSINE (4.000/o): Building block of blood antibodies, strengthens circulatory system and maintains
normal growth of cells.
METHIONINE (2.170/o): Vital lipotropic (fat and lipid metabolizing) amino acid that maintains liver health.
An anti-stress factor, it calms the nerves.
PHENYLALANINE (3.950/o): Required by the thyroid gland for production of thyroxine which stimulates
metabolic rate.
THREONINE (4.170/o): Improves intestinal competence and digestive assimilation.
TRYPTOPHANE (1.1301o): Increases utilization of B vitamins,improves nerve health and stability of the
emotions. Promotes sense of calm.
VALINE (6.0001o): Stimulates mental capacity and muscle coordination.
Non-essential amino acids
ALANINE (5.820/o): Strengthens cellular walls.
ARGININE (5.98%): Important to male sexual health as seminal fluid is 80 percent arginine. Also helps
detoxify the blood.
ASPARTIC ACID (6.340/o): Aids transformation of carbohydrates into cellular energy.
CYSTINE (0.670/o): Aids pancreatic health, which stabilizes blood sugar and carbohydrate metabolism.
Has been used to alleviate some symptoms of food allergy and intolerance. >
GLUTAMIC ACID (8.940/o): With glucose, one of the principal fuels for the brain cells. Has been used to
reduce the craving for alcohol and stabilize mental health.
GLYCINE (3.5%): Promotes energy and oxygen use in the cells.
HISTIDINE (1.08%): Strengthens nerve relays, especially in the auditory organs. Has been used to reverse
some cases of deafness.
PROLINE (2.970/o): A precursor of glutamic acid.
SERINE (4.00%): Helps form the protective fatty sheaths surrounding nerve fibers.
TYROSINE (4.60%): Slows aging of cells and suppresses hunger centers in the hypothalamus. Can be
synthesized from phenylalanine. Involved in proper coloration of hair and skin, including protection from
sunburn.

Minerals:
1. POTASSIUM (15,400 mg/kg): A crucial mineral that regulates body electrolyte balance. Deficiency can
cause heart arrest, hypertension, adrenal exhaustion and muscular collapse.
2. CALCIUM (1,315 mg/kg): The most abundant mineral in the body, it is especially important to bone and
dental health, but is also involved in neural transmissions to the muscles. Spirulina supplies about as
much calcium, gram for gram, as milk.
3. ZINC (39 mg/kg): The pivot point of over thirty vital enzymatic reactions, with profound effects on mental
health, skin tone, prostate function and healing capacity.
4. MAGNESIUM (1,915 mg/kg): Deficiency can lead to spasmodic muscle disorders, including cardiac
irregularities. Helps assimilation of vitamin C, B vitamins and protein.
5. MANGANESE (25 mg/kg): Activates enzyme systems, along with zinc. Promotes activity of
neurotransmitter acetylcholine, and helps stabilize blood sugar.
6. SELENIUM (0.40 ppm): Originally believed to be a toxic heavy metal, but now known to be necessary for
health. It retards aging, harmful oxidation and free radical formation, reduces the toxic effect of
carcinogens, and improves cardiac efficiency.
7. IRON (580 mg/kg): Promotes formation of hemoglobin, the oxygen-carrying blood pigment found in
healthy red blood cells. Iron deficiency is most common among women in their reproductive years.
8. PHOSPHORUS (8,942 mg/kg): The second most abundant mineral in the human body, it is found in
practically every cell. Functions with calcium to maintain bone density. Helps to digest carbohydrates and
the B vitamins niacin and riboflavin.

Vitamins:
1. PYRIDOXINE or B6 (3 mg/kg): Involved in breakdown and assimilation of protein. Protects cardiac health,
reduces edema and stabilizes female hormone levels. Dr. Carl Pfeiffer has demonstrated that B6,
together with the mineral zinc, can cure some forms of schizophrenia.
2. BIOTIN (0.4 mg/kg): An enzyme that carries CO, during certain biochemical reactions involved in
carbohydrate metabolism. Also acts as a co-enzyme in the assimilation of other B-complex vitamins.
Biotin is destroyed by eating raw egg whites and some kinds of raw fish.
3. COBALAMIN or B12 (2 mg/kg): The most difficult of all vitamins to obtain from vegetable sources. A B12
deficiency results in pernicious anemia, nerve degeneration, premature senility, pronounced fatigue and
mental illnesses resembling schizophrenia.
4. PANTOTHENIC ACID (11 mg/kg): The "stress" vitamin, used by the adrenal glands, along with cholesterol
and vitamin C, to manufacture cortisone and other steroids in response to physical and mental stress.
Deficiency encourages sensitivity to allergy, infection and degenerative diseases such as arthritis and
rheumatism. Ulcers and hypoglycemia have also been associated with shortage of this vitamin.
5. FOLIC ACID (0.5 mg/kg): Essential to proper hemoglobin formation in red blood cells. Deficiency results in
anemia, poor growth, skin pigmentation disorders and premature graying of the hair.
6. INOSITOL (350 mg/kg): Vital lipotropic nutrient that sustains liver health and helps detoxify carcinogens,
particularly excess female hormones. Helps normalize blood cholesterol levels. With choline, inositol is
used by the liver to manufacture lecithin. Inositol is the second most abundant vitamin in the body, after
niacin. Recent studies indicate that inositol, with biotin, reduces loss of scalp hair.
7. NIACIN (118 mg/kg): Also known as nicotinic acid and niacinamide, which is an alternative form, niacin is
essential to mental health. Dr. Abram Hoffer, a renowned pioneer in orthomolecular psychiatry, has
completely relieved schizophrenic symptoms using niacin. The Physicians' Desk Reference, a
pharmaceutical text used by doctors when prescribing medication, recognizes niacin as an effective
cholesterol lowering agent.
8. RIBOFLAVIN or B2 (40 mg/kg): The most common vitamin deficiency is that of riboflavin and results in
cataracts, failing vision, watery eyes and uncontrollable eczema.
9. THIAMINE or B 1 (55 mg/kg): A co-enzyme in the breakdown of dietary carbohydrate. Maintains levels of
glucose in the blood. Deficiency results in weakness, cardiac damage, abdominal distention and poor
oxygenation. Severe shortage results in death; critical toxemia develops from unmetabolized
carbohydrate fragments.
10. TOCOPHEROL or vitamin E (190 mg/kg): Spirulina contains more vitamin E per gram than pure wheat
germ. This nutrient protects heart and vascular health, promotes oxygenation of cells, and retards aging.

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Carotenoids:
Alpha-carotene -- traces
Beta-carotene -- 1,700 mg/kg
Xanthophylis -- 1,000 mg/kg
Cryptoxanthin -- 556 mg/kg
Echinenone -- 439 mg/kg
Zeaxanthin -- 316 mg/kg
Lutein -- 289 mg/kg

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Enzymatic pigments:
Chlorophyll
Phycocyanin
Porphyrin
phycoerythrin
tetrapyrrole
phytonadione

An analysis of the components of Spirulina and Chlorella (in 100g)

Nutrients

Spriulina

Chlorella

Protein

55 - 80g

55 - 60g

Lipid

5 - 7.5g

10 - 12g

Chlorophyll

1 - 2.5g

1 - 2g

Carotenoid

0.2 - 0.4g

0.2 - 0.4g

Phycocyanin

2.5 - 9.5g

0g

Potassium

1,100 - 2,200mg

1,000 - 1,400mg

Calcium

500 - 700mg

200 - 400mg

Phosphorus

340 - 720mg

300 - 500mg

Magnesium

250 - 700mg

250 - 300mg

Fe

50 - 100mg

50 - 80mg

Pro-Vitamin A

20 - 30mg

0.4 - 0.5mg

Vitamin B1

2 - 4mg

1 - 2mg

Vitamin B2

3 - 5mg

4 - 5mg

Vitamin B6

0.8 - 1.2mg

0.8 - 1.2mg

Vitamin B12

0.2 - 0.3mg

0.1 - 0.2mg

Vitamin E

8 - 10mg

6 - 8mg

Nicotine Acid

8 - 10mg

8 - 10mg

Inositol

70 - 180mg

80 - 100mg

COMPARISION OF SPIRULINA WITH OTHER FOOD PRODUCTS

Vitamin content

Equivalent proportion

Vitamin A (Beta-carotene/ Retinol)

225 apples or 14 glasses of milk

Vitamin B1 (Thiamine)

4 bowls of rice or 8 sardines

Vitamin B2 (Riboflavine)

2 potatoes or 210 grapes

Vitamin B3 (Niacin)

5 oranges

Vitamin B12 (Cyanocobalamin)

120 grams of cheese

Vitamin E (Alpha Tocopherol)

7 glasses of milk or 2 chicken

Amounts of protein in food products (%)

Protein

Spriulina

Soybeans

Beef

Egg

Fish

65 - 70%

34 .3%

19.3%

12.7%

20.0%

Amount of essential amino acids in Food (%)

Isoleucine
Leucine
Lysine
Methionine
Cystine

Spirulina
3.25-3.9
5.9-6.5
2.6-3.3
1.3-2.0
0.5-0.7

Chlorella
5.5
7.7
5.7
1.7
0.4

Soybeans
1.8
2.7
2.58
0.43
0.48

Beef
0.93
1.7
1.76
0.43
0.23

Egg
0.67
1.08
0.89
0.43
0.35

Fish
0.83
1.28
1.95
0.58
0.38

FAO
Standard
4.2
4.8
4.2
2.2
4.2

Phenylalanine
Tyrosine
Threonine
Tryptophane
Valine

2.6-3.3
2.6-3.3
3.0-3.6
1.0-1.6
4.0-4.6

4.1
1.5
4.3
1.1
4.9

1.98
1.38
1.62
0.55
1.86

0.86
0.68
0.86
0.25
1.05

0.65
0.49
0.59
0.2
0.83

0.61
0.61
0.99
0.3
1.02

2.8
----2.8
1.4
4.2

AZOLLA

Experiments have shown that the salt-resisting limit of Azolla filiculoides is 0.7% (salt water) and its
alkali-resisting limit is 0.3% (Na 2 CO 3 + NaHCO 3 ).
Raises Tacon (1989) that aquatic fern (dry basis) can be used in shrimp feed and fish, as it reveals a
good nutritive value: crude protein: 25.3, crude fat: 3.8, crude fiber, ash 9.3 and 12.5

Santiago et al. (1988), El-Salle (1992) included in diets for tilapia the Azolla sp 10% growth and
reached levels above the control treatment, but when included in diets of Azolla sp on a 25% growth
and reached values lower than the control treatment.
These aquatic plants can be used as feed for farm animals and fish (Than et al. 1997) because they
are sources of high nutritional value protein (18 to 32% PB) but have a deficiency that are too
voluminous for their food low dry matter production (5 to 6%) (Bytniewska et al., 1980).
Gopal (1987) said that they should be analyzed in the high water content of the food, suggesting a
drying treatment to reduce the volume of the inclusion or for silage which adds a little system. For
the above we recommend the possibility of a better use of these foods in smaller species of farm or
freshwater fish, which are promising in terms of high yields of biomass.

CHLORELLA

DUNALEILLA

Co-culture of microalgae with shrimp.

The previous sections have demonstrated that intake by the shrimp of various algal-derived
compounds and metabolites can enhance its immune system and resistance to bacterial and viral
pathogens. These generally entailed the recovery of the compounds through various extraction
procedures and their administration to shrimp via injection, immersion and ingestion modes.
Meanwhile, studies have also shown that even just the presence of microalgae in the rearing water
of shrimp and fish can also enhance the survival, growth and transformation index of the cultivated

species (Muller-Feuga 2000). Pond water that is rich in microalgae and associated aggregates
composed of detritus and microorganisms have been reported to improve shrimp growth as well as
its immune response compared to those reared in clear water (Moss et al. 1992; Primavera et al.
2000). On the other hand, microalgal growth can be supported by the excess dissolved nutrients
released by shrimp into the pond water. This mutually beneficial relationship of microalgae and
shrimp resembles that of a natural ecosystem where energy and nutrients can be recycled
efficiently. As the shrimp industry is continually beset with disease problems, co-culture of shrimp
with microalgae provides an alternative approach to farming that is environment-friendly and
sustainable. One of the farming technologies based on the co-culture of shrimp and microalgae is
the greenwater culture system. The term greenwater is generally defined as water that is greenish
in color owing to the dense number of microalgae that is usually enhanced through fertilization
(Moriarty 1997). In shrimp farming, greenwater development is sustained by the presence of finfish
which releases dissolved inorganic nutrients necessary for the growth of microalgae. Production of
greenwater in shrimp farming is either done on an adjacent pond unit or in the shrimp rearing pond
itself with finfish held in a pen-like enclosure. Tilapia, milkfish and grouper are the finfishes
commonly integrated into culture system with the salinetolerant male hybrid of Tilapia hornorum
and Oreochromis niloticus (so-called Jewel tilapia) being the most popular (Tendencia et al. 2004;
Tendencia et al. 2006; Cruz et al. 2008). Figure 1 shows the culture management practices being
adopted in the greenwater culture system based on Corre et al. (2000). It is possible for this culture
system to be made biosecure which is essential in excluding WSSV. This is by minimizing or
completely eliminating the need for seawater addition into the finfish rearing unit. Water released
from the shrimp rearing unit can also be directed towards the finfish rearing unit in order to provide
additional dissolved nutrients to the microalgal component.
(http://www.trnres.com/ebook/uploads/caipangivcontent/T_1437039455Caipang%204.pdf)

RECOMMENDATION:

3716 L/ pond of 200 sft WSA is estimated to yield 9.290 Kg Dry Spirulina in 5-7 days with
controlled release of nutrients.
That is to say, 1 Ha WSA is estimated to yield 150 Kg Dry Spirulina in 5-7 days with controlled
release of nutrients.

Mother algal culture contains

Thalassiosira Weissfloggii
Spirulina platensis
Nostoc ellipsoporum
Dunaliella tertiolecta
Chlorella pyrenoidosa
Azolla filiculoides
Anabaena sp.

ECONOMICS
Let us take 5 Ha shrimp rearing ponds
1 Ha algal pond is required.
Algal pond water Column Height: 1 Foot
Algal pond water Volume = 3x10^6 L
Inoculum needed: 12 L / Ha pond
Medium Nutrients/Day: 6 Kg/day

Suggested Replacement of Artificial Feed with live natural algal Feed:

From Day 1 to Day 30: Use only 45% of recommended artificial feed
From Day 30- 60 Days Use 55% recommended artificial Feed
From Day 60-90 Days Use 65% recommended artificial Feed
From Day 90-110/120 Days: Use 75% recommended artificial Feed
Caution: Monitor Feed by checking check trays, feed consumption, body weights of the
shrimp, health of the shrimp, climatic changes etc.

Expected Savings in Total feed: 30 to 40%

Expected Savings per Piece of Shrimp: Rs 0.75 to Rs 1.00
Expected Savings per Kg of Live Shrimp: Rs 22.50 to Rs 30.00
Further, better pigmentation, better disease resistance, better FCR, better
daily body weight gain, better survival rate.
Web References:
http://www.oilgae.com/non_fuel_products/aquaculture.html
https://www.youtube.com/watch?v=USkN8Ah2CzE&feature=youtu.be

REFERENCES:
1.

2.
3.
4.
5.
6.
7.
8.

Anonymous, 1991. The design and operation of live feeds production systems. In: Rotifer and
micro-algae culture systems, Fulks, W. and Main K.L. (Eds.). Proceedings of a US-Asia Workshop,
Honolulu, Hawaii, January 28-31, 1991. The Oceanic Institute, Hawaii, USA, pp 3-52.
Barnab, G. 1990. Harvesting micro-algae. In: Aquaculture, Volume I. Barnab, G. (Ed.). Ellis
Horwood, New York, pp 207-212.
Bolsunovskii, A.Y. & Kosinenko, S.V. 2000. Intracellular phosphorus pool of the cyanobacterium
Spirulina platensis. Mikrobiologiya (Microbiology-Moscow), 69: 135137.
Brown, M.R. 1991. The amino acid and sugar composition of 16 species of microalgae used in
mariculture. Aquaculture, 145: 79-99.
Brown, M.R. and Miller, K.A. 1992. The ascorbic acid content of eleven species of microalgae used
in mariculture. J. Appl. Phycol., 4: 205-215.
Brown, M.R., Jeffrey, S.W. and Garland, C.D. 1989. Nutritional aspects of microalgae used in
mariculture; a literature review, CSIRO Marine Laboratories Report 205, 44 pp.
Cano, R., Raudez, S. & Hooker, E. 2004. The natural diet of Apocyclops panamensis at a shrimp farm
on the Pacific coast of Nicaragua. Zoological Stud., 43: 344349.
Chang, Z.Z., Zhu, W.B., Ye, M.X., Fang, Y. & Zhang, J.Y. 1999. The possibility of nitrifying bacteria
inoculation in Spirulina mass culture. Jiangsu J. Agril. Sci., 15: 191192.

9.
10.

11.

12.

13.

14.
15.
16.
17.

18.

19.

20.
21.

22.
23.

24.

25.

26.
27.
28.

Costa, J.A.V., Colla, L.M. & Filho, P.F.D. 2004. Improving Spirulina platensis biomass yield using a
fedbatch process. Bioresource Technol., 92: 237241.
Coutteau, P. and Sorgeloos, P. 1992. The requirement for live algae and their replacement by
artificial diets in the hatchery and nursery rearing of bivalve molluscs: an international survey. J.
Shellfish Res., 11(2): 467-476.
Coutteau, P., Hadley, N., Manzi, J. and Sorgeloos, P. 1994. Effect of algal ration and substitution of
algae by manipulated yeast diets on growth of juvenile Mercenaria mercenaria. Aquaculture, 120:
135-150.
De Pauw, N. and Persoone, G. 1988. Micro-algae for aquaculture. In: Micro-algal Biotechnology.
Borowitzka, M.A. and L.J. Borowitzka (Eds.). Cambridge University Press, Cambridge, U.K., pp 197221.
Donaldson, J. 1991. Commercial production of microalgae at Coast Oyster Company. In: Rotifer and
microalgae culture systems, Proceedings of a US-Asia Workshop, Honolulu, Hawaii, January 28-31,
1991. Fulks, W. and K.L. Main (eds). The Oceanic Institute, Hawaii, USA, pp 229-236.
El-Sayed, A.F.M. 1994. Evaluation of soybean meal, spirulina meal and chicken offal meal as protein
sources for silver seabream (Rhabdosargus sarba) fingerlings. Aquaculture, 127: 169176.
Fox, J.M. 1983. Intensive algal culture techniques. In: CRC Handbook of mariculture. Volume 1.
Crustacean Aquaculture. McVey J P (Ed.). CRC Press, Inc., Boca Raton, Florida, USA, pp 43-69.
Fox, R.D. 1985. Spirulina, the alga that can end malnutrition. The Futurist, 19: 3035.
Gladue, R. 1991. Heterotrophic microalgae production: Potential for application to aquaculture
feeds. In: Rotifer and microalgae culture systems, Proceedings of a US-Asia Workshop, Honolulu,
Hawaii, January 28-31, 1991. Fulks, W and K.L. Main (eds). The Oceanic Institute, Hawaii, USA, pp
276-286.
Helm, M.M., Laing I., and Jones, E. 1979. Culture of algae for larval fish and shellfish rearing. Part 1.
The development of a 200 l algal culture vessel at Conwy, Fisheries Research Technical Report,
MAFF Direct. Fish. Res., Lowestoft (53), UK, 18 pp.
Herrero, C., C. Angeles, J. Fabregas, and J. Abalde 1991. Yields in biomass and chemical constituents
of four commercially important marine microalgae with different culture media. Aquacult. Eng., 10:
99-110.
Howell B.K. & Matthews, A.D. 1991. The carotenoids of wild and blue disease affected farmed tiger
shrimp (Penaeus monodon, Fabricius). Comp. Biochem. & Physiol., 98B: 375379.
Kaji, T., Okabe, M., Shimada, S., Yamamoto, C., Fujiwara, Y., Lee, J.B. & Hayashi, T. 2004. Sodium
spirulan as a potent inhibitor of arterial smooth muscle cell proliferation in vitro. Life Sci., 74: 2431
2439. Kebede, E. & Ahlgren, G. 1996. Optimum growth conditions and light utilization efficiency of
Spirulina platensis (= Arthrospira fusiformis) (Cyanophyta) from Lake Chitu, Ethiopia. Hydrobiol.,
332: 99109.
Laing, I. 1991. Cultivation of marine unicellular algae. MAFF Laboratory Leaflet Number 67.
Directorate of Fisheries Research Lowestoft, UK. 31 pp.
Lee, C-S and Tamaru, C.S. 1993. Live larval food production at the Oceanic Institute, Hawai. In: CRC
Handbook of mariculture. Volume 1. Crustacean Aquaculture, 2nd Edition. McVey J.P. (Ed.). CRC
Press, Inc., Boca Raton, Florida, USA, pp 15-28.
Liao, I., Su, H.-S. and Lin, J.-H 1993. Larval foods for penaeid prawns. In: CRC Handbook of
mariculture. Volume 1. Crustacean Aquaculture, 2nd Edition. McVey J.P. (Ed.). CRC Press, Inc., Boca
Raton, Florida, USA, pp 29-59.
Liao, W.L., Nur-E-Borhan, S.A., Okada, S., Matsui, T. & Yamaguchi, K. 1993. Pigmentation of cultured
black tiger prawn by feeding with a Spirulina-supplemented diet. Bull. Japanese Soc. Sci. Fish.
(Nippon Suisan Gakkaishi), 59: 165169.
Lu, J. & Takeuchi, T. 2004. Spawning and egg quality of the tilapia, Oreochromis niloticus fed solely
on raw Spirulina platensis throughout three generations. Aquaculture, 234: 625640.
Maeda, S. & Sakaguchi, T. 1990. Accumulation and detoxification of toxic metal elements by algae.
Introduction to Appl. Phycol., 109136.
Nguyen, H.T., Nguyen, T.C., Dang, D.K. & Dang, H.P.H. 1980. The first results of investigations and
cultivation of Spirulina platensis in Vietman. Rev. Hydrobiol. Bulg. Acad. Sci., 9. Okamura, H. &
Aoyama, I. 1994. Interactive toxic effect and distribution of heavy metals in phytoplankton. Toxicol.
& Water Quality, 9: 715. Olgun, E.J. 1986. Appropriate biotechnology systems in the arid
environment. In H.W. Doelle, & C.G. Heln, eds. Applied Microbiology. Dordrecht, D. Reidel Publ.
Com., Paris, UNESCO, Trends in Sci. & Res., 2: 111134.

29. Palanisamy, V., Latif, F.A. and Resat, R.B.M. 1991. A guide on the production of algal culture for use
in shrimp hatcheries. National Prawn Fry Production and Research Centre, Pulau Sayak, Kedah,
Department of Fisheries, Ministry of Agriculture, Malaysia. 23 pp.
30. Phang, L.Y. Kun, M.A. Borowitzka & B.A. Whitton, eds. In Proc. 1st Asia-Pacific Conference on Algal
Biotechnology. Kuala Lumpur, Malaysia. University of Malaya. Britton G., Armitt, G.M., Lau, S.Y.M.,
Patel, A.K. & Shone, C.C. 1981. Carotenoproteins. In Carotenoid Chemistry & Biochemistry. G.
Britton and T. W. Goodwin, eds. pp. 237251. Oxford. Pergamon Press. Britz, P.J. 1996. The
suitability of selected protein sources for inclusion in formulated diets for the South African
abalone, Haliotis midae. Aquaculture, 140: 6373.
31. Sheldon, R.W. and Parsons T.R. 1967. A practical manual on the use of the Coulter counter in
marine research. Coulter Electronics Sales Company, Ontario, Canada, 65 pp.
32. Smith, L.L., Fox, J.M. and Granvil, D.R. 1993a. Intensive algae culture techniques. In: CRC Handbook
of mariculture. Volume 1. Crustacean Aquaculture, 2nd Edition. McVey J.P. (Ed.). CRC Press, Inc.,
Boca Raton, Florida, USA, pp 3-13.
33. Smith, L.L., Fox, J.M. and Granvil, D.R. 1993b. Intensive larviculture techniques. In: CRC Handbook
of mariculture. Volume 1. Crustacean Aquaculture, 2nd Edition. McVey J.P. (Ed.). CRC Press, Inc.,
Boca Raton, Florida, USA, pp 153-172.
34. Spoehr, H.A. & Milner, H.A. 1949. The Chemical Composition of Chlorella: Effect of Environmental
Conditions. Plant Physiol. 24: 120. Stanier, R.Y., & Van Niel, Y. 1962. The concept of a bacterium.
Arch. Mikrobiol., 42:1735.
35. Toyomizu, M., Sato, K., Taroda, H., Kato, T. & Akiba, Y. 2001. Effects of dietary Spirulina on meat
color in muscle of broiler chicken. British Poultry Sci., 42: 197202.
36. Tredici, M.R., Papuzzo, T. & Tomasello, L. 1986. Outdoor mass culture of Spirulina maxima in seawater. Appl. Microbiol. Biotechnol., 24: 4750.
37. Utting, S.D. and Spencer, B.E. 1991. The hatchery culture of bivalve mollusc larvae and juveniles.
MAFF Laboratory Leaflet Number 68. Directorate of Fisheries Research, Lowestoft, UK. 31 pp.
38. Venkataraman, L.V., Somasekaran, T. & Becker, E.W. 1994. Replacement value of blue-green alga
(Spirulina platensis) for fish meal and a vitamin-mineral premix for broiler chicks. British Poultry
Sci., 3: 373381.
39. Volkman, J.K., Jeffrey, S.W., Nichols, P.D, Rogers, G.I. and Garland, C.D. 1989. Fatty acids and lipid
classes of ten species of microalgae used in mariculture. J. Exp. Mar. Biol. Ecol., 128: 219-240.
40. Vonshak, A. (ed.). 1997. Spirulina platensis (Arthrospira). In Physiology, Cell Biology and
Biotechnology. Basingstoke, Hants, London, UK, Taylor and Francis. Vonshak, A. & Guy, R. 1988.
Photoinhibition as a limiting factor in outdoor cultivation of Spirulina platensis. In Stadler et al. eds.
Algal Biotechnology, pp. 365370. London, Elsevier Applied Sci. Publishers. Vonshak, A. &
Richmond, A. 1988. Mass production of the blue-green alga Spirulina: an overview. Biomass, 15:
233247.
41. HEEL, A.G.J. (1995): Application of nutrient data under Practical Conditions Requirement: Special
Problems of intensive fish farming systems. J. Appl. Ichthyol. 11: 205-214.
42. Thanh Hang, D., Van Lai, N.; Lylian, R. and Ly, J. 1997. Digestion and nitrogen metabolism in Mong
Cai pigs fed sugar cane juice and foliages diference as source of protein. Livestock Research for
Rural Development (9) 2: 17 25.
43. VAN HOVE, C. AND LOPEZ, Y. 1983. In Azolla physiology. Technical Bulletin, National University of
Colombia, Faculty of Agricultural Sciences, Palmira. Volume I, Number 1. p 43 58.
44.