The present invention relates to development of a commercial process for the culture of the micro algae
Spirulina (Arthrospira platensis) in a natural way. The culture media used in this process contain only
natural ingredients of organic nature. No synthetic chemicals have been added in the culture media directly
or indirectly. The yield of organic spirulina was 5 to 6 g/m2/day by this process. The quality of spirulina
product obtained was considerably in a better quality compared to the available commercial spirulina in
terms of protein, chlorophyll, and other vitamins, minerals. The present invention describes the organic
process for the manufacture of spirulina in commercial quantities.
Prior Art:
Many process for the manufacture of spirulina has been proposed, by growing the said alga in an aqueous
medium comprising various carbon sources along with sodium chloride, di potassium hydrogen phosphate,
sodium nitrate, potassium sulphate, magnesium sulphate, calcium chloride, ferric EDTA, trace elements,
urea or ammonium chloride and potassium chloride. Carbon source is selected from group consisting
bicarbonate, carbonate, acetate, glucose and sucrose or combination of thereof.
A process to produce spirulina biomass using a source of development of said microalgae from rejects from
the alcohol industry, notably sugar vinase and carbondioxide originating from' fermentation vessels have
been given.
An invention related to a method of culturing spirulina algae for production of enhanced biomass and
cellular ingredients using the medium comprising of organic carbon source have been dealt with.
Spirulina biomass which is rich in iron, algae rich in gamma linolenic acid (GLA), linoleic and arachidonic
acids, essential amino acids, vitamins, minerals and chlorophyll process for the preparation of phycocyanin
a natural blue colorant from spirulina species were given in the prior art works.
None of the prior art dealt with usage of organic oil cakes as nitrogen source for growing spirulina
culture.
There is no prior art defined for the fermentation of organic inputs using anaerobic micro organism
for release of nitrogen from oil cakes for the growth of spirulina.
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Introduction:
Spirulina (Arthrospira platensis) is a blue-green microscopic cyanobacterium plant that has existed on our
planet for 3.6 billion years and served as a source of nourishment to ancient civilizations. It has been
consumed by human and animals. Spirulina has been commercially produced worldwide using chemical
ingredients (nonorganic spirulina) or using only naturally obtained ingredients (Organic spirulina). Spirulina
has a full spectrum of nutrients that nourish both the body and the mind. Organic Spirulina is grown in an
environment that guarantees the highest level of purity, use of only naturally organic grown ingredients as
input materials as culture media for growing spirulina, making it more easily digestible and pleasant tasting
than other forms.
An optimum source for antioxidant protection that will boost your immune system healthy and vital. Organic
spirulina ranks as one of nature's most powerful "Superfoods." At over 60% protein, spirulina is the most
potent and digestible source of vegetarian protein. This family member of blue-green algae packs in a
whole lot of antioxidant and phytonutrient nutrition along with high amounts of beta-carotene, essential
amino acids, minerals, B vitamins, and chlorophyll. Spirulina is also rich in gamma-linolenic acid (GLA).
Research suggests spirulina's potential benefits for promoting total body health, including…
 Promote normal immune response
 Support optimal digestion and weight management
 Promote optimal cardiovascular health
 Provide vital energy for the body and cognitive brain function
 Support heavy metal detoxification
 Promote optimal gastrointestinal flora
As none of the prior art describe the complete natural / organic process for the commercial manufacture of
spirulina, herewith we propose a commercial process for the manufacture of spirulina algae using a culture
media which contain only the naturally available mineral and vegetable oil cakes as ingredients. The
essential nutrients required for the growth of the algae is supplied only by organic inputs. Synthetic
chemicals have been completely avoided in the process, so that the algae obtained in our process will be
not only organic and completely naturally grown but also possess the benefit of same commercial outcome
through optimized yield and quality of the product in terms of protein, chlorophyll and beta carotene.
DEFINITIONS
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Unless defined otherwise, all technical and scientific terms used herein have the same meaning as
commonly understood by one of ordinary skill in the art to which this disclosure belongs.
In describing the present invention, the following terms will be employed, and are intended to be defined as
indicated below.
For purpose of this invention, the term “OD” means "optical density." It is a method of measuring how many
organisms in a sample is by measuring its turbidity. More cells would absorb more light. Putting the sample
into a spectrophotometer measures its absorbance and value is recorded as OD.
For purposes of this invention, “TA” mean “Total Alkalinity”. Total alkalinity is a measure of the total amount
of dissolved particles in the solution whose pH is higher than 7.0.
For purpose of this invention, the term pH defines the acidity or alkalinity of a solution on a logarithmic
scale on which 7 is neutral, lower values are more acid and higher values more alkaline.
For purpose of this invention, the term TDS defines the total dissolved solids in a solution.
Process of Cultivation of Organic Spirulina
Microorganism and culture medium of microalgae Spirulina platensis, strain was used as inoculums. The
culture mediums were made in commercial ponds of 500m2 and 1000m2 surface areas. The ponds are
provided open sun light provision, with paddle-wheels used to agitate the water.
Nutrients needed for culture media:
The following ingredients were used in the culture media for developing organic spirulina in commercial
ponds. All ingredients were obtained from natural sources / organic grown through certified suppliers. No
synthetic chemical entity has been used in this process of production.
1. Protein source:
Spirulina contains an unusually high amount of protein ranging from 60% and 65% by dry weight,
containing all essential amino acids. Spirulina is rich in essential fatty acids such as gamma-linolenic acid,
and also provides alpha-linolenic acid, linoleic acid, stearidonic acid, eicosapentaenoic acid,
docosahexaenoic acid, and arachidonic acid. The source of aminoacids and proteins for spirulina is
provided in the culture by using oil cakes of a mixture of several fixed oils such as soyabeen oil, castor oil,
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ground nut oil, neem oil, pungam oil, sesame oil, sun flower oil etc in a definite ratio ranging from 10 to
90%.
2. Fermentation:
The input oil cakes were fermented using effective microorganisms containing a mixture of Lactobacilli
cesai, Rhodopseuodomonas, yeast and photosynthetic bacteria for a period of 7 to 15 days at 25 to 35odeg
C in anaerobic conditions without sunlight.
3. Carbon:
The organic Spirulina is provided with a rich source of carbon and from natural organic oil cakes of
vegetable oils. This helps in biosynthesis of chlorophyll, vitamins and other nutrients by the spirulina
through photosynthesis using sunlight.
4. Bicarbonate (HCO3):
Natural sodium bicarbonate obtained as pure baking soda made from the deposits of mineralized sodium
bicarbonate extracted by a simple water process without using any chemicals has been used in the culture
media for organic spirulina. The mineral needs and alkalinity required for the growth of spirlina algae has
been provided by the natural bicarbonate ions.
5. Sodium chloride and Minerals:
The organic Spirulina is provided with a rich source of potassium, calcium, chromium, copper, iron,
magnesium, manganese, phosphorus, selenium, sodium, and zinc from natural sea salt.
Steps involved in Organic Process of spirulina:
 Initially we need to charge water and innoculum into spirulina pond
 Charge fermented organic inputs in specified quantities
 During inoculmn the parameters were set as pH was ranging from 8 to 10. HCO3
concentration was 1.0 to 3.0g/L; TA was 2.0 to 4.0g/L and the OD values ranges from 0.4
to 1.0
 On regular production of spirulina, the daily monitoring paratemetrs were maintained as :
The pH was ranging from 9 to 11. HCO3 concentration was 0.2 to 0.8g/L; TA was 2.0 to
4.0g/L and the OD values ranges from 0.4 to 1.0
 After 3 days based on the analysis data, add the fermented inputs once again
 Continue analysis every day
 After a week, if necessary add fermented inputs again
 After 15 days of innoculum, harvest the spirulina (OD should be more than 0.7)
 After every harvesting, add the fermented inputs to the pond
 Harvest once in six days.
 The slurry of organic spirulina is washed with water
 The washed slurry is spray dried to get the dry powder of organic spirulina.
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Yield:
An average of 5 to 6 g/m2/day of organic spirulina is obtained as dry powder.
EXAMPLES:
Example 1
 Oil cakes of soyabeen oil 10%, castor oil 40%, neem oil 15%, pungam oil 5%, sesame oil
5% were mixed with water and stirred continuously in a anaerobic tank fitted with a
mechanical stirrer
 Effective microorganism mixture containing Lactobacilli cesai, Rhodopseuodomonas, yeast
& few photosynthetic bacteria has been added to the mixture of oil cakes for initiating
fermentation of the protein nutrients by the microorganisms
 After about 7 to 10 days of fermentation, the supernatant liquid which is enriched in free
nitrogen is transferred to another tank in anaerobic condition and kept ready for use in
culture media
 Spirulina culture tanks of 500m2 surface area in which organic spirulina is to be grown are
cleaned well and filled with water having TDS less than 500 ppm up to 11.5cm depth in
the tank.
 Add 200kg of natural sodium bicarbonate and 100kg of sea salt in to the tank.
 From the 2nd fermentation tank, 1000L of clear liquid enriched with nitrogen is transferred
through pipelines in to the spirulina tanks
 The spirulina inoculums developed in lab were added in to the tank and mechanical stirring
of the tank contents were carried out using an agitator in the tank to oxygenate the
contents
 The culture was allowed to grown in natural sun light under continuous agitation.
 Observations were made on daily basis for the parameters such as microscopy of cell
morphology, OD, pH, HCO3, Total alkalinity and water depth in the tank.
 The nutrient content in the tank such as protein content, phosphates, micro elements and
carbonates were estimated on a routine basis and enough natural nutrient ingredients
were supplied in a regular intervals
 The culture was allowed to grown initially for 15 days from the inoculums day for obtaining
a good to medium small coils of spirulina with OD >1.0
 Harvesting the organic grown spirulina were carried out through filtration, followed by
washing the thick slurry of spirulina with sufficient water
 The water washed thich slurry of spirulina with TDS about 10% were dried in spray drier to
obtain dry powder of spirulina in dark green color
 The product was packed in sterile conditions in bags and analysed for the target
specifications
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 Yield of spirulina was 5.0 to 5.5g/m2/day
Example 2
 Oil cakes of soyabeen oil 10%, ground nut oil 40%, neem oil 15%, pungam oil 5%, sun
flower oil 5% were mixed with water and stirred continuously in a anaerobic tank fitted with
a mechanical stirrer
 Effective microorganism mixture containing Lactobacilli cesai, Rhodopseuodomonas, yeast
& few photosynthetic bacteria has been added to the mixture of oil cakes for initiating
fermentation of the protein nutrients by the microorganisms
 After about 7 to 10 days of fermentation, the supernatant liquid which is enriched in free
nitrogen is transferred to another tank in anaerobic condition and kept ready for use in
culture media
 Spirulina culture tanks of 500m2 surface area in which organic spirulina is to be grown are
cleaned well and filled with water having TDS less than 500 ppm up to 11.5cm depth in the
tank.
 Add 200kg of natural sodium bicarbonate and 100kg of sea salt in to the tank.
 From the 2nd fermentation tank, 1000L of clear liquid enriched with nitrogen is transferred
through pipelines in to the spirulina tanks
 The spirulina inoculums developed in lab were added in to the tank and mechanical stirring
of the tank contents were carried out using an agitator in the tank to oxygenate the
contents
 The culture was allowed to grown in natural sun light under continuous agitation.
 Observations were made on daily basis for the parameters such as microscopy of cell
morphology, OD, pH, HCO3, Total alkalinity and water depth in the tank.
 The nutrient content in the tank such as protein content, phosphates, micro elements and
carbonates were estimated on a routine basis and enough natural nutrient ingredients
were supplied in a regular intervals
 The culture was allowed to grown initially for 15 days from the inoculums day for obtaining
a good to medium small coils of spirulina with OD >1.0
 Harvesting the organic grown spirulina were carried out through filtration, followed by
washing the thick slurry of spirulina with sufficient water
 The water washed thich slurry of spirulina with TDS about 10% were dried in spray drier to
obtain dry powder of spirulina in dark green color
 The product was packed in sterile conditions in bags and analysed for the target
specifications
 Yield of spirulina was 5.0 to 6.0 g/m2/day
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Quality control parameters of the produced organic spirulina were analysed and optimized as below.
PARAMETER UNIT SPECIFICATION TEST RESULTS
(On dry basis)
ANALYSIS METHOD/
REFERENCE
Moisture % NMT- 8 % 7.4% AOAC Official method 930.04,
16th Edn.
Protein % NLT- 60 % 62.46% AOAC Official method 978.04,
16th Edn.
Ash % NMT- 9 % 5.71% AOAC Official method 930.05,
16th Edn
Bulk Density
50 Tapped
gm/cc NLT- 0.5 0.766 gm/cc US Pharmacopeia. USP Chapter
616 Bulk Density and Tapped
Density. 2006:2638.
Chlorophyll A % 0.8 - 1.2 1.07 % Vonshak.A.1997. Spirulina
platensis (Arthrospira)
physiology, cell biology and
biotechnology.
Beta Carotene μg / gm 1600 - 2200 1870μg / gm Ranganna .S. (1986). Handbook
of analysis and QC for fruit &
veg. Products
Crude
Phycocyanin
% 9 - 13 11.04 % Boussiba S. and A. Richmond.
1979. Isolation and purification
of phycocyanins from the bluegreen
alga Spirulina platensis.
Arch. Microbiol. 120:155-159.
Total Heavy
Metals
PPM NMT -5 Complies AOAC 18th Edn. : 2006
Pb PPM NMT- 0.5 3.34 PPM AOAC 18th Edn. : 2006
As PPM NMT- 0.2 0.059PPM AOAC 18th Edn. : 2006
Hg PPM NMT- 0.05 0.039 PPM AOAC 18th Edn. : 2006
Cd PPM NMT- 0.2 0.49PPM AOAC 18th Edn. : 2006
Total Plate Count CFU/gm NMT- 50,000 48210Cfu/gm
Total Yeast and
Mould
CFU/gm NMT- 100 Less than
10Cfu/gm
Salmonella /25 gm Absent Absent
Shigella /25 gm Absent Absent
E.Coli /25 gm Absent Absent
Staphylococcus /25 gm Absent Absent
Coliforms /25 gm Absent Absent
USFDA, Bacteriological
Analytical manual 3rd, Edn,

We claim:
1. An organic method for production of spirulina, the method comprising steps of:
a) mixing oil cakes as a nitrogen source with water and stirring continuously in an anaerobic tank;
b) adding microorganisms to the oil cakes for fermentation of oil cakes which helps for the
release of nitrogen to the culture and helps for protein synthesis in the algae;
c) maintaining a temperature between 25OC to 35OC for a time duration of 7 to 15 days in
anaerobic conditions without sunlight leading to fermentation of the organic contents and
release the nitrogen available for spirulina;
d) transferring the supernatant liquid to second tank in anaerobic condition;
e) transferring the supernatant liquid from second tank to the spirulina tanks;
f) adding spirulina inoculums to the spirulina tank;
g) stirring the contents of spirulina tank in presence of natural sun light for 15 days till an optical
density of the culture reaches the value of 0.7 to 1.0;
h) observing the parameters such as microscopy of cell morphology, OD,pH,HCO3, total
alkalinity and water depth in the tank on the daily basis;
i) observing and maintaining nutrient ingredients in the spirulina tank at a regular interval;
j) harvesting the spirulina from the spirulina tank;
k) washing the spirulina with water and
l) drying the spirulina of step (k) to obtain dry powder of spirulina.
2. The organic method for production of spirulina as claimed in claim 1, wherein anaerobic tank is
fitted with a mechanical stirrer.
3. The organic method for production of spirulina as claimed in claim 1, wherein the oil cakes as a
nitrogen source are selected from the group of soyabeen oil 10%, castor oil 40%, neem oil,
pungam oil 5%, sesame oil 5%, sunflower oil 5%, ground nut oil 40%.
4. The organic method for production of spirulina as claimed in claim 1, wherein the microorganism
mixture contain Lactobacilli cesai, Rhodopseudomonas, yeast and few photosynthetic bacteria.
5. The organic method for production of spirulina as claimed in claim 1, wherein the fermentation
period ranges from 1 to 10 days.
6. The organic method for production of spirulina as claimed in claim 1, wherein the spirulina tank of
step e) of claim 1 having a surface ranging from 500m2 to 2500m2
7. The organic method for production of spirulina as claimed in claim 3, wherein the spirulina tank is
filled with water having TDS less than 500ppm with a depth ranging from 10 to 20 in the tank.
8. The organic method for production of spirulina as claimed in claim 3, wherein the spirulina tank is
enriched with natural sodium bicarbonate and sea salt.
9. The organic method for production of spirulina as claimed in claim 1, wherein the parameters
maintained in the pond during inoculumn were pH ranging from 8 to 10; HCO3 (bicarbonate
concentration) from 1.0 to 3.0g/L;TA (total alkalinity) ranging from 2.0 to 4.0g/L.
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10. The organic method for production of spirulina as claimed in claim 1, wherein the parameters
maintained in the pond during regular production as HCO3 (bicarbonate concentration) from 0.2 to
0.8g/L; pH ranging from 9 to 11; TA (total alkalinity) ranging from 2.0 to 4.0g/L.
11. The organic method for production of spirulina as claimed in claim 1, wherein the harvesting of
organic spirulina was carried out when the OD (Optical density) values were from 0.4 to 1.0
12. The organic method for production of spirulina as claimed in claim 5, wherein the amount of natural
sodium bicarbonate is 200kg and sea salt is 100kg.
13. The organic method for production of spirulina as claimed in claim1, wherein the spirulina is
harvested through filtration, followed by washing the thick slurry of spirulina with sufficient water.
14. The organic method for production of spirulina as claimed in claim 1, wherein the spirulina of step
(k) of claim 1 is dried by spray drying.
15. The organic method for production of spirulina as claimed in claim 1, wherein the yield of spirulina
is 5.0 to 6.0 g/m2/day.