Hi all, I'm new in this forum. I'm a post grad student from Malaysia. I have some questions to ask you guys, about growing Bb.

Just bought myself culture of Bb from UK (CCAP) and USA (UTEX). I'm maintaining the cultures in the 3N Modified Bold Medium. It has been almost a month, it doesn't grow as fast as I thought (I feel I'm lucky already coz it is still alive). I'm growing it in 500ml conical flasks. I do not know how to measure the light intensity, as I use the normal aquarium light to provide luminescents in 12 hours photoperiod basis.

And also, I've found that there are whitish fungi-like thing growing in the medium as well, and not sure how to get rid of it. I think i will try expose the medium to the sun and see whether the UV light kills those things. Will that work?

The third thing is that I'm wondering why some of the Bb colonies sink at the bottom of the flask and some of them are floating on the surface.

I don't pump CO2 to the culture, does CO2 aeration make any different? Does Bb likes to stay stationary or we have to keep them shake on the shaker?

How to measure their growth rate? According to the literature, Bb double every 2-3 days right? Is measuring the dry weight is the best approach to determine their growth rate? But it kills the algae though... is there any non-invasive method?

 

If CO2 does matter, I plan to use CO2 from yeast fermentation since I do not have enough money to buy the CO2 tank and regulator those costly stuff. Is that ok?

  

Aeroplane,

 

The easy way to measure growth is with a Stecci(SP?) disk.  It is a black dot about 14mm across with a white dot about 6mm in the middle of it.  It is attached to a stick about 36 cm long. 

 

Put a stick-on ruler, or markings each CM or sonething up the stick, so you can measure depth.

 

Dip the disk into your tank and keep lowering it until you can no longer see the disk.  At that point, read the depth. 

There are calibrations for various species of algae, but you will just need to record your readings to see how much your BB is growing.  (If your first reading is 20cm and your second is 10cm, you have doubled.)

 

Try to read it in the same light conditions each day - same time of day, same windows open, same lights on, etc. 

 

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I think you need air bubbling through the growth medium - a small aquarium air pump, some tubing, a splitter, and a couple of airstones.  That will supply the algae with CO2, take away O2, provide a little movement, etc.

 

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Patrick reported that his algae liked the CO2 from fermentation. 

What temperature are you growing the Bb at? salinity? Ph? 12 hr photoperiod is good, how strong is the light and in what colours? There's a way to measure the population using equipment originally developed for counting blood cells. I'll post more information later. Right now, I am off to MIT for that Energy Club meeting I posted about.

THank you guys, I will try to make the suggested changes and see how it work. Will keep you guys informed.

Hi all, this is the picture that i took from my culture. What are those granular stuffs in the cell? Is that the oil that we are going to extract?

Hi Bobk, I am growing it at an average of 27-28 C. pH neutral, 12hr photoperiod. I use normal aquarium light, slightly blue in color.

Aeroplane,

 

It would be my guess that those are the oil globules.  The nice think about BB is that we don't have to extract them.  They will migrate outside the cell, where we can collect them and let the BB cells continue to grow and produce more oil.

 

I assume you are aware that the oil is very different from a typical plant oil.  It is more like crude oil, just cleaner.  It is believed that you cannot transesterify it, that you must crack it like crude.

 

 

Thank you liberty for your information. That helps a lot.  

I bought Bb from different places, and there are different isolation numbers as well. Some of them tends to clump together, some of them appears in yellow orange, some of them are like greenish... Basically I'm trying to ask why they are so different, even under the microscope.

 

Base on what i've observed in the last 3 months, my Botryococcus braunii grows better on the solid agar medium compare to the liquid medium. Probably because I didn't pump CO2 into the liquid.

Aeroplane,

I really liked the picture.  It's got me thinking of getting a microscope with digital camera.  What was the setup you used? What magnifacation did you need?

Ken

Hi Ken,

We don't need to have those luxurious digital camera equipped microscopes to take the pictures. All you need is a normal 5 mega pix digicam and stick the camera to the eye piece of the microscope. Then you can take down the picture. Remember, don't shake your hand because it is very sensitive to any movement.

:)

http://item.express.ebay.com/__Microscopes-Parts_40x-1600x-BINOCULAR-MICROSCOPE-DIGITAL-USB-PC-CAMERA_W0QQitemZ170094662977QQihZ007QQptdnZMicroscopesQ20Q26Q20PartsQQptdiZ795QQcmdZExpressItem

 We have a unit like this budgeted for July 1. Cant wait to hook it up the comp/monitor and zoom around.

Using liquid nitrogen to freeze dry the algae biomass consumes a lot of energy. I would like to know if I dry it under rtp, will the oil evaporated as well?

Aeroplane,

At rtp, oil doesn't evaporate.

Depending on what you are doing, you may be able to skip the drying.  It is need for test quantities to get accurate lipid percentages, but may not be needed for other processes.

 

 

hi, i'm italian. Sorry to my english.

Hi Teki, You will need to buy the culture media and culture them :)

UTEX provides the recipe for culture media as well. You can refer to their website.

Good luck to you~

 I have been facing this problem.

The smaller "algae" obviously does not look like Bb. I don know what are those things. These smaller algae does not clump together like Bb. After some time (about 1 month) in the culture medium, it become the dominant culture in the medium. I don't think this is contamination because I did a negative control. I can't spot any of these in negative control.

I'm sure the stock culture is clean as well, it is from UTEX. I also check it under the microscope before i innoculate them. What are these little green algae?

 

Aero,

Maybe they are baby BB.  Since the BB will clump, maybe you could seperate some of the unkowns and put them into a separate culture.  If they grow up to be BB, all is well.

 

 

 

Thanks liberty. I have recultured the \'baby\'. Any other suggestion what would it be?

Aero,

I'm afraid I'm not much help.