Hi,
i am working on dynamic of cyanobacteria in a lake, and i want to do a culture and isolate a specie of cyanobacteria specifically Cylindrospermopsis, to do some experiments, what is the right method to isolate and which medium should i use ? please
cordially
Hello Seridi Lamia
1. collect your sample from the location in a sterile falcon and incubated it in your algal incubator (a fluorescence lamp (2000 ± 200 Lux) with 12:12 h dark/light cycles at ambient temperature for 2-7 days to adapt)
2. Serial dilution in test tube to try to purify your sample and incubate under the same conditions for 15 days until biomass appears (green or blue-green color, depending on your samples)
3. start to swap it on agar BG11 plates and let them grow. Then examine the colonies, choose the purified one, and transfer it to a new sterilized tube or container, allowing it to grow.
We used BG11 media for growing cyanobacteria.
I attached two articles, one of them written by Bolch et al., and it was a reference to teach me how I culture cyanobacteria.
Article Algal-Derived Synthesis of Silver Nanoparticles Using the Un...
genetic, morphological, and toxicological variation among globally distributed strains of Nodularia (Cyanobacteria)
Hi Seridi,
Cylindrospermopsis is a heterocyte-forming genus. So, if this genus is the focus of your study and you do not care about other non heterocytous genera (e.g. Oscillatoria, Phormidium, Leptolyngbya, etc), I would recommend BG110 (Same recipe as BG11 but without nitrogen source). In this way you will get rid of many genera that do not fix nitrogen and will make the isolation process a bit easier.
When it comes to isolation, I recommend this book chapter to you:
Rippka R. Isolation and purification of cyanobacteria. Methods Enzymol. 1988;167:3-27. doi: 10.1016/0076-6879(88)67004-2.
Briefly, the basic idea is to spread your crude material on the plate, incubate, then choose single filaments and transfer those on fresh plates. Depending on your sample and experience, it may take multiple transfers until you manage to get unicyanobacterial cultures so you need to be very patient and keep trying until you succeed!
best of luck with your experiments!
Maria
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