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Why the pH value decreased seriously in my algal culture?

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#1 biobunny



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Posted 04 March 2009 - 03:24 AM

Hi Is there anyone who would be so kind as to give me some idea to explain this phenomenon?

I cultured several flasks of one green microalgae species in TAP medium under irradiance of 90 umol.m-2.s-1,
I added 18g/L glucose in the medium, and after 9days of cutlure, I found the aglal culture in bad growth condition though the biomass accumulated was much higher than the control culture without glucose, the color of the algae showed a tendency of turning white!! I collected the cells and tested the pH value of the culture medium, and it was below pH4.5!! I think most of the algal cells must have died! The pH value in the control culture without glucose was 8.0. I checked the culture under microscope and no bacteria was found.

I think the whole thing is very confusing :unsure:

The nitrogen source in TAP medium is NH4Cl, which should bring the pH down upon algal growth. So this could be a possible reason for the pH decrease in the above culture. But in another culture which also contained 18g/L glucose and under a much stronger irradiance of 800 umol.m-2.s-1. , the pH was 7.0 and with even more biomass accumulated!!! If NH4Cl is the criminal, then why the culture under stronger irradiance which should have taken in much more NH4Cl to produce biomass did not give a even lower pH??

Is it possible that the CO2 produced by respiration can cause a pH as acid as below 4.5?

The following is TAP recipe:

Tris - Acetate - Phosphate (TAP) Medium

(modified from Durnford Lab)

For One Liter of Medium:

2X Filnerís Beijernicks Solution 25ml

1M Potassium Phosphate 1ml

Trace mineral solution or 5ml

P IV solution 2X (see SVM) 3ml

Tris-Base 2.42g

Glacial Acetic Acid (17.4 mM acetate) 1ml

(pH should be 7.2)

Stock solutions:

2X Filnerís Beijernicks Solution (500 ml)

NH4Cl 8g

CaCl2 * 2H2O 1g

MgSO4 *7H2O 2g

Trace Mineral Solution (500 ml)

5 g disodium EDTA Ė dissolve in 400 ml water by heating and stirring

Neutralize to pH 6.5 with 5N NaoH

Add each of the following in order. Allow each to dissolve completely before adding the next.

0.5 g FeSO4*7H2O

2.2 g ZnSO4*7H2O

1.14 g H3BO3

0.51 g MnCl2*4H2O

0.016 g CuSO4*5H2O

0.073 g Na2MoO4*2H2O

0.016 g CoCl2*6H2O

1M Potassium Phosphate Stock (50 ml)

add: 20 ml 1M stock KH2PO4 (1M stock: 6.8g/50ml)

30 ml 1M stock K2HPO4 (1M stock: 8.7g/50ml)

pH 7.2 at RT

#2 molgen



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Posted 05 March 2009 - 02:35 AM

CO2 can and dos lower the pH.
Try a bottle with a 22u filter for passive diffusion of gas.

#3 pito



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Posted 07 March 2009 - 10:35 AM

Hallo biobunny,

as molgen allready said: CO2 can indeed lower the pH.
CO2 is often used in watersystems to lower the pH.

you might wanna check the following: http://www.lenntech....bon-dioxide.htm

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.

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