Experiment to test for the better growth medium for

spirulina

Procedures
1. Separate the spirulina from the initial medium by filtration.
2. Wash the spirulina using 2.5% NaCl solution to remove residue of the original solution.
3. Prepare 300 ml of BG-11 medium, SSD1+2 medium and 2.5% NaCl solution respectively.
4. Through weighing by difference, add 0.02 g (fresh mass) of spirulina to each solution with a
spatula.
5. Obtain a sample of each culture to be put under the microscope in a haemocytometer to
ensure that the biomass concentration is similar
6. Complete the set up by adding the air supply, a cleaned stir bar and a watch glass as cover
for each set up
7. Every two day, extract a sample of each set up, dilute the sample with 2.5% NaCl solution (if
needed), and measure then record the biomass concentration.
8. Calculate and record the specific growth rate.

*Note:
The pH of SSD1+2 is 8.3, while the pH of BG-11 is 7.2.
Spirulina prefers an alkaline environment, within the range of pH 8.0-11.0.

The sudden drop in biomass concentration of the BG-11 medium suddenly is possibly due to the
fact that the spirulina trichomes adhere to the wall of the beakers for growth, a phenomenon
similar to that observed in the previous samples grown in the BG-11 medium prior to the
experiment.

On 29/6, the NaCl solution remains clear while the SSD1+2 medium is visibly greener than the
rest, and in the BG-11 medium, yellow solids, potentially dead spirulina, can be observed,
similar to that observed in the previous samples grown in the BG-11 medium prior to the
experiment.

Materials
Materials BG-11 Medium Broth Powder1

1
BG-11 Medium Broth Powder, optimized for cyanobacteria, produced by JC Teaching Aids Company,
Hong Kong; Product code: USB-BG11H; Website:
http://www.jc-teachingaids.com/eshop/index.php?route=product/product&product_id=419
 Spirulina plate sis dry grow medium SSD1+22
NaCl 50g
Distilled water 2 L
Spirulina (Arthrospira platensis)
Filter paper

Apparatus Beakers x 3
Stir bars x 3
Air pump with air outlet x 3
LED light
Electric balance
Microscope
Haemocytometer
Slide cover
Watch glass x 3
Dropper x 3
Spatula x 4

Set-up

Data

Biomass concentration (no. of trichomes per mm2)

2
Spirulina plate sis dry grow medium SSD1+2, produced by HealthAlgae, Sweden; SKU:
A-S003-0800; Website:
https://www.healthalgae.com/product/spirulina-platensis-80-l-dry-grow-medium-ssd12/
 Date 25/6 26/6 29/6 30/6 2/7 3/7 5/7 6/7
(9:00) (9:00 (9:00) (10:0 (10:0 (9:00 (9:00) (9:00)
) 0) 0) )

BG-11 0.778 0.88 0.000 0.000 0.00 0.000 0.000 0.111

9 0

SSD 0.667 0.77 2.222 2.333 1.77 8.222 5.333 8.667

1+2 8 8

NaCl 0.222 0.33 0.000 0.000 0.00 0.000 0.000 0.000

3 0

Specific growth rate

Where, N0 is the population size at the beginning of a time interval, Nt is the population size at
the end of the time interval.3

Date 25/6-26/6 29/6 30/6 2/7 3/7 5/7

BG- 0.00556 NA NA NA NA NA
11

SSD 0.00641 0.0146 0.00195 -0.00566 0.0666 -0.00902

1+2

25% 0.0169 NA NA NA NA NA
NaCl

Conclusion and explanation

The SSD1+2 medium is proven to be the superior growth medium in terms of sustaining the
growth of spirulina.

3
Ranjith, L., Shukla, S. P., Vennila, A., & Purushothaman, C. S. (2013). Growth performance of Spirulina
(Arthrospira) platensis in a low cost medium: An assessment. Acta Biologica Indica, 335–342.
 A sound explanation is that it has provided the culture with an alkaline environment which the
BG-11 medium has failed to create. The initial pH of the SSD1+2 medium measured is 8.3 while
that of the BG-11 medium is 7.2 and that of the 2.5% NaCl solution is 7.1. The most suitable pH
for spirulina lies between 8.5 and 10.0, a range which both secures the growth of spirulina and
prevents infections. Normally, as the culture continues to grow, the pH of the solution will
increase as carbonic acid present in the solution will be absorbed by the culture for
photosynthesis. However, in a low initial biomass concentration (about 0.07 g/L for each setup),
in a environment that is initially less alkaline than desired (as in the case of the batch grown in
the BG-11 medium), the culture is unable to grow in a rate that is satisfactory and enables the
spirulina to bring about an increase in the pH of the solution. Therefore, the pH of the BG-11
medium and the NaCl solution remains lower than ideal, (final pH of BG-11 medium: 6.8; final
pH of NaCl: 6.6; decrease in pH value of solution is most probably caused by a rate of
respiration that is higher than the rate of photosynthesis) limiting the growth of the culture. The
pH of the SSD1+2 medium, on the other hand, continuously increases, reaching a final pH of
10.1, providing an ideal growing environment for the spirulina culture.

Error and solution

Evaporation may lead to an increase in biomass concentration or the concentration of
nutrients, leading to inaccuracy in measurement of biomass concentration and a higher growth
rate than normal respectively. To cancel out the impact, addition of water to maintain the volume
of solution in each setup at 300 ml is carried out.

Formation of colonies of trichomes or adhesion of trichomes to the wall may affect the biomass
concentration measured with a haemocytometer. To minimize the impact, air is introduced into
the solution with an air pump to stir and aerate the solution.