Spirulina growth medium
A special setup of nutrients are needed to grow Spirulina. This page will list supplies you need to grow Spirulina plus the recipes and formulas – simple and complex – that you need to grow Spirulina (and also other algae or cyanobacteria)
Sections on this page about Spirulina growth medium
List of Spirulina growth mediums
There are a number of Spirulina growth mediums that have been developed today. Here is a compiled list:
– Zarrouk’s medium
–
– …to be continued
Our complete Spirulina growth medium ready to mix with water (tap water works very well as long as it doesn’t contain chlorine). It is based on the Zarrouk’s medium but has been adapted more to the proportions of nutrient that Spirulina actually need. E.g. extra nitrogen (and only nitrate) to give higher yields per liter. For a 10 liter culture where you harvest regularly (10% daily) this medium will cover you for at least 2 months. Mix it with water, stir for circa 20 minutes and add to your culture. Alternatively you can add it direct to the culture with the water. You can vary the concentration from 0.5x to 3x and still grow well.
Recipe for Zarrouk's medium
Zarrouks medium is one of the most popular Spirulina growings mediums used in research.
Components used in Zarrouk's medium
Components (PART A - Zarrouk's medium)
Gram per Liter
(g/L)
Gram per Gallon
(GPG)
Molarity (mmole/L)
NaHCO3 (Sodium Bicarbonate)
16.80
63.58
Na & C: 199.98
K2HPO4 (Dipotassium Phosphate)
0.50
1.89
K: 5.74
P: 2.87
Components (PART B - Zarrouk's medium)
Gram per Liter
(g/L)
Gram per Gallon
(GPG)
Molarity (mmole/L)
NaNO3 (Sodium Nitrate)
2.50
9.46
Na & N: 29.42
K2SO4 (Potassium Sulfate)
1.00
3.79
K: 11.48
S: 5.73
NaCl (Sodium Chloride)
1.00
3.79
-
MgSO4·7H2O (Magnesium Sulfate Heptahydrate)
0.20
0.76
-
EDTA-Na2·2H2O (Disodium Ethylenediaminetetraacetate Dihydrate)
0.08
0.30
-
CaCl2·2H2O (Calcium Chloride Dihydrate)
0.04
0.15
-
FeSO4·2H2O (Iron(II) Sulfate Heptahydrate)
0.01
0.04
-
Components (PART C - Zarrouk's medium)
Trace elements mixture A:
10 ml to 1 liter of PART A + B
Gram or mg per Liter (g/L) or (mg/L)
Gram or mg per Gallon
(GPG) or (mgPG)
Molarity (mmole/L)
H3BO3 (Boric acid)
2.86 mg
10.83 mg
-
MnCl2·4H2O (Manganese Chloride Tetrahydrate)
1.810
6.85
-
ZnSO4·7H2O (Zinc Sulfate Heptahydrate)
0.222
0.84
-
MoO3·0.015 (Sodium Molybdate)
0.015
0.057
-
CuSO4·5H2O (Copper Sulfate Pentahydrate)
0.074
0.28
-
Components (PART D - Zarrouk's medium)
Trace elements mixture B:
1 ml to 1 liter of PART A + B
Gram or mg per Liter (g/L) or (mg/L)
Gram or mg per Gallon
(GPG) or (mgPG)
Molarity (mmole/L)
NH4VO3 (Ammonium Metavanadate)
22.9 mg
86.7 mg
-
NiSO4·7H2O (Nickel Sulfate Heptahydrate)
47.8 mg
180.9 mg
-
NaWO2 (Sodium Tungstate Dihydrate)
17.9 mg
67.8 mg
-
Ti2(SO4)3·6H2O (Titanium(III) Sulfate Hexahydrate)
4.4 mg
16.7 mg
-
Co(NO3)2·6H2O (Cobalt(II) Nitrate Hexahydrate)
4.4 mg
16.7 mg
-
Source: Amara & Steinbüchel 2013. 1 gallon is approximately equal to 3.78541 liters.
How to prepare the Zarrouk's medium
It is not stated in the source (Amara & Steinbüchel 2013), but it can be assumed that you scale the components in Part A and Part B, add them into a beaker or suitable vessel, and then add deionized/distilled water. You do this during mixing until the total volume reaches 1 liter (if you are making 1 liter of Zarrouk’s medium).
The other way is to add the components in Part A and B to 1 liter of deionized/distilled water. However, that will make the final volume above 1 liter in total.
For Part C, you do the same as above, and for Part D, also the same. Then as the last step, when you are going to use the Zarrouks medium, you add 10 ml from Part C and 1 ml from Part D per total liter of Part A and B.
For sterile cultivation, you first autoclave the prepared Part A and B mixture, followed by sterile filtration of Part C and D as they are added. If you autoclave the final mixture of all parts, it is likely that some of the ions will adhere to each other and form strong and not soluble salts that fall out on the bottom of the medium mixture. This makes the components unusable by the Spirulina cells when you cultivate.
Since you are using so little of Part C and D, it is good to aliquot the rest into 10, 50, or 100 ml volumes and freeze them until you need them again. That way, you keep them fresh and avoid extra work.
Zarrouk's medium raw data
If you want the Zarrouk’s medium as raw data:
One liter of Zarrouk’s medium consists of (part A) NaHCO3 16.80 g and K2HPO4 0.50 g; (part B) NaNO3 2.50 g, K2SO4 1.00 g, NaCl 1.00 g, MgSO4·7H2O 0.20 g, EDTA-Na2·2H2O 0.08 g, CaCl2·2H2O 0.04 g, and FeSO4·2H2O 0.01 g; trace elements mixture A (part C 10 mL/l): 1.00 mL, trace elements mixture B (part D 1.0 mL/l): 1.00 mL; part C mg/L: H3BO3 2.86, MnCl2·4H2O 1.810 g, ZnSO4·7H2O 0.222 MoO3·0.015, and CuSO4·5H2O 0.074 (the used amount is 10 mL/l); part D mg/L: NH4VO3 22.9, NiSO4·7H2O 47.8, NaWO2 17.9, Ti2(SO4)3·6H2O, and Co(NO3)2·6H2O 4.4 (the amount used was 1.0 mL/l) (Amara & Steinbüchel 2013).
Figure. The proportions and how to mix the different parts of the Zarrouk’s medium.
NPK ratio and mineral concentrations of Zarrouk's medium
From the the list above we can get the following molarities (mmol/L) :
– Nitrogen:
– Phosphorus:
– Potasium:
HealthAlgae's medium vs Zarrouk's medium
HealthAlgae’s SSD1+2 medium is very similar to the Zarrouk’s medium but has been improved to fit the ratio of nutrients needed by the Spirulina cells.
Emphasis has been placed on providing more nitrogen since this is the nutrient depleted before any of the others.
The nitrogen also only comes in nitrate (NO3-) form in HealthAlgae’s medium. This enables higher nitrogen concentrations to be used in comparison to ammonia/ammonium (NH4/NH3+), which is toxic in very small amounts.
However, it can be recommended to add ammonia/ammonium or urea (which breaks down to ammonia/ammonium and CO2) on a regular low feed to further improve a dense and fast-growing Spirulina culture. But that requires that temperature, mixing, pH, and foremost light are providing enough to yield even higher densities. The latter are often the limiting factors and not the medium nitrogen.
HealthAlgae’s Spirulina growth medium also comes in the form of one dry mix and is ready to be added to tap or mineral water (21 grams per liter), mixed, and then used directly to grow Spirulina.
You can find HealthAlgae’s Spirulina growth medium here if you wish to order:
Components (Zarrouk's medium)
Gram per Liter
(g/L)
Gram per Gallon
(GPG)
Molarity (mole/L)
NaNO₃ (Sodium Nitrate)
1.5
5.7
K₂HPO₄ (Dipotassium Phosphate)
0.02
0.08
MgSO₄·7H₂O (Magnesium Sulfate)
0.075
0.284
CaCl₂ (Calcium Chloride)
0.036
0.136
NaCl (Sodium Chloride)
0.01
0.038
Na₂CO₃ (Sodium Carbonate)
0.001
0.0038
H₃BO₃ (Boric Acid)
0.0003
0.0011
MnCl₂·4H₂O (Manganese Chloride)
0.0016
0.0061
ZnSO₄·7H₂O (Zinc Sulfate)
0.00022
0.00083
MoO₃ (Sodium Molybdate)
0.00005
0.00019
CuSO₄·5H₂O (Copper Sulfate)
0.000036
0.00014
CoCl₂·6H₂O (Cobalt Chloride)
0.00002
0.000076
FeSO₄·7H₂O (Ferrous Sulfate)
0.00002
0.000076
EDTA (Ethylenediaminetetraacetic Acid)
0.1
0.38
pH adjusted to 9.5 using NaOH
-
-
Source: Amara & Steinbüchel 2013
References
- Amara, A., & Steinbüchel, A. (2013). New Medium for Pharmaceutical Grade Arthrospira. International Journal of Bacteriology, 2013, PMC4745477. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4745477/
How to find a Spirulina culture
Component Amount per Liter Amount per Gallon
NaNO₃ (Sodium Nitrate) 1.5 g 5.7 g
K₂HPO₄ (Dipotassium Phosphate) 0.02 g 0.08 g
MgSO₄·7H₂O (Magnesium Sulfate) 0.075 g 0.284 g
CaCl₂ (Calcium Chloride) 0.036 g 0.136 g
NaCl (Sodium Chloride) 0.01 g 0.038 g
Na₂CO₃ (Sodium Carbonate) 0.001 g 0.0038 g
H₃BO₃ (Boric Acid) 0.0003 g 0.0011 g
MnCl₂·4H₂O (Manganese Chloride) 0.0016 g 0.0061 g
ZnSO₄·7H₂O (Zinc Sulfate) 0.00022 g 0.00083 g
MoO₃ (Sodium Molybdate) 0.00005 g 0.00019 g
CuSO₄·5H₂O (Copper Sulfate) 0.000036 g 0.00014 g
CoCl₂·6H₂O (Cobalt Chloride) 0.00002 g 0.000076 g
FeSO₄·7H₂O (Ferrous Sulfate) 0.00002 g 0.000076 g
EDTA (Ethylenediaminetetraacetic Acid) 0.1 g 0.38 g
pH adjusted to 9.5 using NaOH – –
Live Spirulina cultures for sale from farmers
The second option is to go to a Spirulina grower near you. It is possible that they can sell you their Spirulina culture and you can start growing your own Spirulina from that one. Just be sure to get the specific nutrients they use too. This is good since the farmer might use a specific nutrient setup that the culture is more adapted too and if not followed will result in low growth in the start.
The large and best option to get a steady Spirulina culture going with 10x concentrated 500 ml Spirulina start culture and a complete Spirulina manual.
Great starter of your own culture with 500 ml 10x concentrated Spirulina culture, full instructions manual and 10 L dry complete growth medium ready to mix with water and use.
Thank you
– What to do when you get you have ordered your Spirulina starter culture
– Direct steps to take when you get your live Spirulina culture
– How to maintain the Spirulina culture from HealthAlgae when you get it
– Setting up a high yielding Spirulina culture system
If you are interested in how to make an photobioreactor or read about the ones you can order from us, have a look here: Photobioreactors for growing Spirulina and algae : link to the photoreactors
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