Growing Algae...

[Savvypro]

17 June 2011 update - part 2...

I've just uploaded a video of what I can see under my microscope. Its at: Spirulina Platensis 17/June/2011

[Savvypro]

18 June 2011 update...

Nothing much to report on the Spirulina front.

On the other two strains, they look to be doing well, the area of the agar slant that their covering is increasing.

The biofuel algae is starting to grow everywhere, even in the area between the agar and the test tube wall.

The other strain is now green, especially where its covering agar, there are some red sports at the top end of the test tube which is agarless. Those spots look to be the same algae but in their dormant state.

New video showing what I mean: 18 June Update...

[Savvypro]

21 June 2011...

I haven't made an update post since the 18th, as there hasn't been much to report on the algae.

On the equipment side I've received my digital ph meter, digital TDS meter, as well as my 5 stage Reverse Osmosis Unit.

Still need to calibrate the meters, but the readings I’v done on my tap water are: pH: 7.3, TDS: 314 ppm. If the TDS numbers don’t change much after calibration I may just find that I got a pretty sweet deal on my RO unit. Before the readings I was thinking of adding a DI stage and a Zeolite stage, before the 1 micron filter - which then feeds to the membrane. The reason being is that I thought I had higher TDS, and because of the increase in radio active particles - the Zeolite stage should help.

Still on it's way is a digital Lux meter, a non contact IR thermometer and a couple of more things - I think.

My intention is to really document everything I possibly can - within monetary reason. As I don’t have unlimited funds, not even a large amount to work with. So I’m making every penny count.

I've also got my eye on a Trinocular Darkfeild 2000x Microscope - but that may have to wait for a bit - as it's over my budget. If anyone is felling generous and willing to contribute to my brain fund. Or have a spare Trinocular Darkfeild 2000x Microscope doing nothing, send me a pm.

[ashtweth]

Thanks for the update my friend, you have done well what an inspiration to read

Ash

[Savvypro]

Thanks for the update my friend, you have done well what an inspiration to read

Ash

No problems Ash, stay tunes as I'll be getting ready to do some real experiments. Once I’ve divided the samples out and multiplied them a couple of times. It’s easier said then done, simply because of the time it will take. Just waiting on the algae to multiply.

New video at: YouTube - ‪9 Days later...‬‏

The video shows the progress so far - after just 9 days.

[Savvypro]

Biofuel...

For those interested in the biofuel side of things, take a look at the following video: YouTube - ‪Biofuels from Algae Project - Brunswick Community College Center for Aquaculture & Biotechnology‬‏

[Savvypro]

In a different thread I stated that I had found the research on how to grow AFA, and that AFA was my next algae to get (actually it's one of a couple more - one of them being a biofuel algae).

The only issue I have with reviling this type of information and the reason I've held back the names of the two mystery strains I am currently growing: is that their edible strains. But specifically, the conditions they grow in are also those that other algae (which can be toxic or produce toxic by-products), can also grow in. This is what is keeping me from telling all. Someone may follow along and could get hurt as they haven't taken the precautions that I am taking.

For example, all that follows is the precautions I’m taking:

Isopropyl Alcohol (99.7% pure, I got 5 litres of the stuff - enough to start making my own disinfectants and so on - if need be),

SURGICAL FACE MASKS (may be over the top, but I got a good deal on two boxes and until I build my laminar flow hood I’m not taking any chances).

Medical gloves - two types. (I got a good deal on 10 boxes of 100 (1000 gloves total) since using them I’ve found out why they were such a good deal (so far 1 glove has had a hole in one of the tips). Can’t complain though as they were only £10 (total including delivery - about $16) and I happened to order two boxes of Nitrile gloves that day as well).

The containers that the algae’s will grow in are all glass (lab test tubes, lab flasks and mason jars), and can be autoclaved.

I also have a pressure cooker to do the autoclaving, but may have to get another one that is bigger - as some of the jars I have are too big for the one I currently have.

Reverse osmosis filter with extra stages for extra pure water.

A laminar flow hood - which I'll be putting together.

To just grow Spirulina, most of the precautions I'm taking are not needed as Spirulina can grow in medium which other things can't survive in. But for the other strains - that's a whole different issue.

[Savvypro]

28 June 2011 update...

Yesterday was the hottest day of the year here in the UK (or at least it was where I am). I decided to move the algae to the north facing kitchen which is normally cooler than the rest of the house. The result wasn't that much of a difference, so I ended up placing the test tubes in a jar of cool water to try and lower the temp a bit. The Spirulina can handle the high temp, but the other two algae do well at a lower temp.

I've decided that when I divide out the algae I'm going to do a couple of Spirulina tubes with agar. Simply because with the algae grown on a slant, you can tell if it's still alive as the test tube wall above the growing area gets covered with water vapour from photosynthesis.

My main concern at this stage is just to ensure that the algae are established and that I can multiply them out, otherwise I'll have to reorder more samples - which are expensive. So far so good, for the two algae in agar, the Spirulina is a different issue as it's in liquid and going by the colour of the water is somewhat subjective.

[Savvypro]

3 July 2011 update...

Yesterday I made up a bath of media for each strain.

The way I've done it is to have all the mediums with out agar (just in case it goes off) that way I can just take as much as I need, add the agar and steam sterilise.

Which I did yesterday. This morning I found that the test tubes which had agar mediums didn't set - but the 75ml flats did. I made up the agar mediums in the flats then poured the required amount (for the test tubes) into the test tubes.

I’ve got some videos of what I did yesterday (which I still need to edit together). It’s quite boring as it’s basically me washing all the glassware, drying them, weighing the chemicals then adding them into the jars. Heating the medium jars in a bath of hot water to increase the absorption of the chems into the water. Boring really.

This morning I transferred samples into the viable flats and test tubes. And will see what the results are. I’ll be redoing the agar test tubes, maybe adding a bit more agar or maybe doing the agar mediums in one container, then filing the test tubes from that container.

On a different note - anyone have an All American Pressure Canner lying around that they no longer need. As I spent most of my time during the steam sterilising stage (using the pressure cooker) in the hallway (I had distance and a wall between me and it, the pressure cooker was always in sight, it was never left unattended and there was enough water in the pot). Using that thing always gives me the impression that it’s going to blow, even though its a new steel pressure cooker.

[Savvypro]

6 July 2011 update...

The Spirulina is doing well, even the small amount that was left in the small vial (and by small amount - I'm talking droplets here, held back by the water tension) is starting to take hold in it's new medium. For the vial I did as follows: I poured out the contents (about 1ml) into a larger test tube with about 15ml of new medium. Even though this dilution is extreme (15 to 1), the Spirulina is getting darker in the new test tube. All the new test tubes including the medical flat, which holds about 50ml of medium (it's a 75ml bottle) is doing well.

The agar flats look to be doing ok - it's too early to tell.

[ashtweth]

Good to hear from you Bro, thanks for the good news, please drop me an email have some extra info i found you for my friend

Ash

[Savvypro]

@Ash - I sent you a pm...

On a different note:

When I was transferring samples from the agar strain test tubes, over to the new agar flats. I used some plastic inoculation loops that I had bought. I didn't like the fact that each time I do a transfer I would have to dispose them. Luckily I saw a site that was offering what looked like DIY loops. But at a high price. So I ordered some Nichrome wire, which arrived this morning.

To DIY your own inoculation loop, just cut off a piece of wire. Get a thin piece of round metal - I got a screw driver that came with my pH meter. Fold the wire in half (don’t bend the middle bit - just get the two end bits together) and take the two end bits which are now together and clamp them in the chuck end of a cordless drill (the business end of the drill - where the bits go).

Then you take the screw driver and stretch out the wire so that the looped end is being pulled away from the drill by the screw driver. Make sure that you don’t kink the wire - trust me on this as it will give you a wonky loop.

Then all you do is press the drill button so that the chuck starts turning - do it slowly. Keep a hold of the screw driver, don’t remove it from the wire where the loop will end up being. As the chuck turns, the wire will start forming a helix - with a loop at the screw driver end. After a few seconds the helix will have arrived that the loop end.

Now you have to be careful, because as the chuck keeps turning, the wire will start to turn on it’s self - because of the tension that is building up. If you let go of the screw driver, it will go flying off. When to stop, is when you decide that the loop is ready. If the wire does turn on it’s self - stop, it’s done.

This process doesn't take long to make a useable loop. If you go too fast, or just keep pressing the drill button - you'll end up wasting wire and maybe hurting yourself or others.

The only thing I needed to buy was the 3m of 25 gauge Nichrome wire which I found on Ebay for a total of £2.19 (about $3.49 at the time of posting). I have enough wire left over to do about a dozen loops.

I did a test with 25cm of wire, and ended up with a loop that's 11cm long. You'll then need to find a handle to attach the loop to, I'm thinking a pen of some sort - maybe with a cap.

[Savvypro]

12 July 2011 update...

I meant to post this yesterday, but didn't have time.

The following video shows the progress of the small vial sample.

Note that I recorded the video after I had left the sample settle, because when the algae is mixed into the medium it’s quite difficult to determine if there is actually anything there (apart from the slight green tint which you see at the end of the video).

YouTube - ‪Spirulina Platensis - small vial test sample - 11/July/2011‬‏


One week ago (see 3rd July 2011 and 7th July 2011 posts), I poured out nearly all the content of the vial into a larger test tube with over 15ml or so of new medium. I then added new fresh medium to what was left in the vial. As you can see from the video, for a test that shouldn't have worked (based on the papers and research that I have read) it’s not doing too badly.

[Savvypro]

16 July 2011 update...

Just did a new batch of agar medium for the two non Spirulina strains. The test tubes are currently cooling down at an angle to form the agar flat. So far, doing the agar medium in one batch, then pouring to the test tubes - looks to be the best way of doing it.

I’ll probably do the transfers tomorrow, as I need to think about how I’m going to do it. I’ve got an experiment or two that I want to do.

The Spirulina in the large test tubes look ready for the next round of multiplication transfers. The Spirulina in the 75ml medical flat bottle still has some way to go but it’s getting there.

[Savvypro]

24 July 2011 update...

Abbreviation: flat refers to the 75ml medical flat bottles I’m using.

The flat of the red mystery strain, looks to have developed a contamination - so I've done 3 transfers from it's original test tube (which isn't contaminated - well it doesn't look contaminated, but unfortunately I can’t tell using my current microscopes as the lenses aren't that good for anything past 400x especially if you need 1000x to see something clearly).

Anyway, I transferred new samples to 2 test tubes and a flask which had 100ml of agar medium. I have a spare test tube with agar medium in storage as a control.

On the other mystery strain, I’ve also made new transfers, but only because the sample in the flat (which I made about a month ago) is growing very slowly. And I wanted to make sure I have other test tubes of the strain growing as backups to the flat.

Now on to the Spirulina, the vial sample just keeps getting greener. Last week the Spirulina were barely able to cover part of the corner of the vial - when the algae had settled to the bottom. Today, the whole of the vial bottom is being covered.

I transferred about 80% of the contents of one of the 3 new Spirulina test tubes, into a flask with about 200ml (medium level was at the 200ml mark on the flask). It just so happens to be about a 15 to 1 expansion (similar to the test I did with the vial sample test tube). I also did the same transfer to the “vial test tube sample” into another flask with 200ml of new medium. Both test tubes then had 10ml of new medium each added to them.

I then poured all the contents of the remaining new test tube into a flat and added 30 ml of new medium to the flat. I then added 10ml of new medium into the almost completely empty test tube. I’m going to be doing the vial experiment but this time on a test tube.

The flat that I created about a month ago, I poured just over half of the medium into a new flat bottle, then added 30ml of new medium into the new flat. The original flat had 40ml of new medium added to it.

At this rate, in 2 to 3 weeks (about the time it’s taken for the test tubes and the liquid Spirulina flat to turn dark green, so dark that anything behind the test tubes and the flat. Thats black in colour - can’t be seen through the Spirulina) I should be able to transfer the contents of the flasks to about 6L of medium (3L per flask). From there I should be able to start harvesting Spirulina.

I’m still trying to determine what the best container will be for bulk growing of the Spirulina (and the other algae) in a limited space. As it stands I’ve just used glass: be it test tubes, the sample vial, the flats and the flasks. They have all been glass - with the only plastic being the caps. When I started buying the equipment about 2 months ago, I weighed up the pros and cons of using either plastic (BPA issue, with BPA free plastics potentially also leaching other chemicals which could turn out to be harmful down the line) or sticking to glass. For the small volume stage I decided to go with glass - plus I managed to get some bargain glass lab ware.

I’ve managed to find some cheap 10l and 20l water/food storage containers which could be used for bulk growing (brewing DemiJohns for example). Finding the same size containers in glass for a similar price is nearly impossible. There are second hand glass bottles and containers, but I’m not going to take the chance on them. Especially when the algae is to be consumed.

The attached image shows my current inventory - if you will. The colour code is as follows:

Blue dot is for Spirulina.
Red dot is for the red mystery strain.
Green dot is for the green mystery strain.

On a different note.

I’ve noticed that with test tubes, the spirulina settle out much faster than in the 75ml glass medical flat bottles. In fact I’m able to watch the self shading process taking place. Unfortunately I can’t get it on video, not with the cameras that I have anyway.