Phytoplankton Culture Starter Guide

[FUGU]

Invertebrates fed on live phytoplankton will have a higher natural HUFA content and increase their nutritional value to fry.

To maintain and develop your phyto culture you need to set aside an area with a warm stable temperature, aeration for the culture vessels, 1-2 plant bulbs rigged approx. 200mm to the side of the culture vessels, and a timer providing 12-24 hours of light per day. Aim to set up 4-6 separate cultures in case a culture crashes.

If you are new to culturing phytoplankton set up your culture undiluted, add 0.5ml Phyto Feed per 2.5 litres of culture, and leave for the first week to ensure it settles in.

Once the phytoplankton is stable your first new culture can be set up with 1 part mature culture and 1 part new culture water. Add the Phyto Feed liquid nutrient at a rate of 1-2ml per 2.5 litre of culture. Traditionally, ratios of 1 part mature culture and 3 parts new culture water have been successful but with this greater dilution effect you are at greater risk of a culture crash. Be realistic on the volumes of phytoplankton you need and balance this with the rates of dilution used, and the number of cultures you wish to set up.

For further reading look no further than the Plankton Culture Manual.

For a culture 'reactor' go to a major supermarket and buy the 5 litre mineral water bottles at 80-90 pence ea., drill the lid, add a rigid plastic airtube and you're away.

[wordley]

..

[FUGU]

Hi - yes, you should be able to providing it is fresh. If the culture has been refridgerated, or shows signs of settling out, avoid diluting the culture during the first week until it is stable. Add a low level of nutrients to the culture when setting up will aid recovery e.g. 0.1ml Phyto Feed per 500 ml of culture.

Good luck!

[FUGU]

Additional Notes for Isocrysis culture:

For first 24 hrs add sterile salt water to the dish so that the algal cells are submerged and then place under constant light.

Use a sterile swab to gently rub the algal cells from the agar plate. The Iso cells will come off the dish in clumps when you use it. Start with only 500 mls of water and add fertiliser at 0.2ml per 500ml of culture.

Ensure the clumps remain in suspension and avoid letting them stay around the top of the container - leave culture to develop for a week.

If after 1 week the culture has not developed a brown colour just add more fertiliser and leave to grow for another week.

After 2 weeks, if the culture still has not developed a brown colour transfer 300ml of the culture to a fresh vessel, add replacement 200ml fresh salt water and fertiliser. Keep the culture at a maximum of 500ml until the culture is chocolate brown in colour.

[drivewayreef]

hi any idea when the iso disks will be in stock ? :thanks:

[FUGU]

Hi - online shop updated - nine Iso disks in stock this PM

Cheers

FUGU

[FUGU]

Autumn update: have been trying these micron filters to protect the Iso cultures

http://i310.photobucket.com/albums/kk429/copeblob/really_big_icon_micronairfilter1.jpg

And have been getting some good results:

http://i310.photobucket.com/albums/kk429/copeblob/really_big_icon_phyto_culture_iso.jpg

And have been trying out some alternate vessels:

http://i310.photobucket.com/albums/kk429/copeblob/big_icon_phyto_culture_nanno.jpg

Happy culturing!

[drivewayreef]

Hi fugu , glad to see your recovered somewhat:applause: That iso looks very good , I've had no success at all , what's you method for sterilizing ?
Cheers Mike

[FUGU]

Hi Mike

The Iso is still going good with 6 mini cultures after nearly 3 weeks from one of our disk isolates. The original culture has been put aside with illumination and no aeration with just a gentle daily swirl; and remains browner than the image above. For Iso feel free to be paranoid and set it up in its own culture chamber from other phytos, isolate the airpump from other phytos, use micron air filters on all airlines, the water is currently double sterilised - first with shock chlorination/dechlorination and then heat sterilised before use. When I added Micro Algae Grow the other day there was a lot of flocculation which didn't look too good but then the culture settled down by the next day. I'll be looking to upscale the culture over the next few weeks so fingers crossed!

Cheers

Fugu

[S7v7n44]

"For further reading look no further than the Plankton Culture Manual".

Where will I find this manual?

[FUGU]

Hi

Plankton Culture Manual - complete with a variety of culture techniques for both phytoplankton and zooplankton:

http://www.zmsystems.co.uk/index.php?app=gbu0&ns=catshow&ref=books

Cheers

Fugu

[FUGU]

Here's the benefits of using a microscope - the phyto comes in all shapes and sizes! Certainly a useful technique when monitoring new and mature cultures.

Nannochloropsis (small round dots):

http://i310.photobucket.com/albums/kk429/copeblob/big_icon_phytocult_nan.jpg

Isochrysis (very irregular in shape):

http://i310.photobucket.com/albums/kk429/copeblob/big_icon_phytocult_iso.jpg

Tetraselmis (highly motile guys hence the blurring):

http://i310.photobucket.com/albums/kk429/copeblob/big_icon_phytocult_tet.jpg

I'll have to video the Tetraselmis next time.

Cheers

Fugu

[rodders1986]

Hi,

Do you use RO water mixed with salt or tap water mixed with salt. Reason I ask is because I have read phyto uses Nitrates and Phosphates to grow?

Thanks

:)

[FUGU]

Hi Rodders yes, you are correct both Nitrate and Phosphate are the 2 biggest ingredients in common phyto fertilisers and the levels will drop as the culture develops. Both RO and tap water can be used for culture however RO will provide a more consistent starting point. However just because you may start with RO you still need to aim for the stored water, taps, pipes, culture vessels, beakers remain sterile at the point of culture start up. CO2 injection into the phyto culture is one aspect worth looking into to boost phyto growth rates; you'll just have to be sure the pH is not adversely affected.

Cheers

Fugu

[rodders1986]

Hello,

Thanks for the reply. Are you saying it's better to go with RO water? I take it, that it would be right? If using RO, would you use a fertilier to top up necessary nitrate/phosphate?

I am thinking of ordering some stuff for you soon, just trying to work it all out first lol

Sorry for all the questions :o

:thanks:

[FUGU]

Hi Rodders - yes, the RO will be more ideal to start with. You use the fertiliser at start up of the culture and as a weekly feed. The Micro Algae Grow fertiliser is in stock and very effective for phyto growth. Just let me know if you have any questions re the items you need.

Cheers

Fugu

[chew1904]

Fugu

I am about to start culturing some phyto and the more I read the more I'm confused by a couple of things.

The first is what SG do you mix the salt water to? I ask because I've seen several different values on different sites.

The other question is around steralising the water and equipment. The equipment is all cleaned with a mild bleach solution and then a large dose of dechlorinator used to neutralise any residue. But how do you steralise you salt water?


Thanks
Chew

[ainsy]

sorry to jump on your thread, but can I ask what you use as a dechlorinator?

[chew1904]

sorry to jump on your thread, but can I ask what you use as a dechlorinator?

I used dechlorinator (such as stress coat or tetra aquasafe) that you would use to take chlorine and heavey metals out of tap water in freshwater tanks. Have used this method for steralising and cleaning all maner of equipment on my freshwater and marine setups for years.

To steralise a bottle for example I fill it with water and bleach, leave it to stand for a few hours then empty it and rince it well, then fill it up with water and add 4 times the dose on the bottle of dechlorinator and leave to stand for half an hour, then empty and rinse and its good to go.

[FUGU]

Hi chew and ainsy

Variations in the SG of live cultures will vary – just monitor the SG of the stock you receive and work with that to begin with to avoid shocking the culture.

S.G.s for Nanno and Tetra are approx 1.0222 and working really well at the moment, previously tried 1.016

Previously Nanno has also been grown at 1.002 for freshwater projects (freshwater rotifers, Daphnia)

S.G. for Isochrysis up to 1.026

For water sterilisation we’re currently doing shock chlorination with calcium hypochlorite and sodium thiosulphate for dechlorinating, the bonus is that both the culture vessel and water are treated at the same time. Heat sterilisation is used for small volumes and starter cultures with a microwave with 1 litre on high for 7 mins and allowed to cool before adding salt.

Also, to remove scale deposits off the side of culture vessels dilute acid (eg a solution of white vinegar and warm water) can be used, left overnight, and then rinsed and left to air dry.

Cheers

Fugu

[chew1904]

Thaks Fugu

I've read that a lot of people are using chlorine crystals designed for shock treating swimming pools and hot tubs to steralise the water and vessels. They then use a few drops of normal aquarium dechlorinator or even leave it with air bubbling through it for 24 hours to drive the chlorine off.

[FUGU]

Hi chewy

You're on the right track. The recommendation is to use a 53 micron strainer to pour your chlorine solution through to avoid the risk of any undissolved granules falling through. At the gas off stage you can use a chlorine test strip to be doubly sure the chlorine has gone.

Cheers

Fugu

[chew1904]

Thanks Fugu. There has to be a market for a complete kit (and the topups) for phyto culture. No one place seems to sell all the bits I need. Although I have to say the chlorine crystals and tests are the only things I can't find on your site. ZM is the only place I've managed to find the airline filters so far. :thumbsup:

[FUGU]

I know what you mean Chewi - anything is possible just let me know if you need anything in the short time otherwise no doubt these will be listed shortly! I'll have the small disc inline filters in shortly. Cheers,

Fugu

[chew1904]

Great stuff thanks Fugu you've been very helpful. Any chance you'll have the plankton culture manual available soon? They are impossible to get hold of at the minute.

[FUGU]

Hi Chewi - we have a 2-3 week lead time on the next batch and very latest 3rd printing plus other goodies. Having said this I'll chk to see what 2nd editions I've put aside due to scuffs on the cover. I'll update you later.

Cheers

Fugu

[chew1904]

Thanks Fugu! :applause:

[baileybum1234]

FUGU you mentioned using CO2. Could you give any more advice on this matter? I understand it can effect PH. I have a PH probe but would rather use it on the Ca reactor. I have the aquamedic phyto reactor if I was to dose bubbles per min during lights on periods could you advise maybe how many bubbles?

Any other tips would be appreciated also!

Thankyou

J

[FUGU]

Hi J

Good question. It's possible SWM have tried? also one of the new LFS in Sheffield has tried in the past by manualling releasing CO2 and getting a good result; I'll see if they've done more. I would say your reactor is sufficiently narrow for decent light penetration. I'm about to set up 30cm dia reactors and CO2 injection is likely to be needed so they'll be an opportunity to test setups with and without injection.

Cheers

Fugu

[baileybum1234]

Yeah SWM utalise CO2 from what I understand. In the aquamedic photoplankton reactor instruction manual they also go on about it as well as the plankton culture manual its self. Im just looking to chat to people who have tried it really. Sounds like you have some ideas of your own. Do you record your findings online at all?

Thanks!

J

[nemoc]

Laing at MAFF suggests only an addition of 0.5% CO2 of the total of the air supplied is required to optimise the balance of growth and pH stability.[/URL]

[URL="http://www.cefas.co.uk/publications/lableaflets/lableaflet67.pdf"]Cultivation of unicellular marine algae - MAFF - CEFAS (http://www.cefas.co.uk/publications/lableaflets/lableaflet67.pdf)

[baileybum1234]

Thanks nemoc interesting document!

I didnt realise it would be so low (0.5%). Maybe I need to hook up my PH meter and let it run for a while just to get an idea of how much CO2 is needed to keep it around 7.5 - 8.

Shame its not as easy as just 'x number of bubbles' per minuite during lights on then a solenoid to shut off CO2 at lights off period. Or maybe it could be in time.

Many people have CO2 on their Ca reactor a T-off to the plankton reactor would be fairly simple.

J

[nemoc]

LOL If only it was that easy :)

I suspect due to the seawater's buffering capacity it is a fine line between that we can use more than we need with little ill effect as no co2 reactor etc; however I also suspect that as density increases the risk of a crash does so too.

Still its fun finding out :)

[FUGU]

Hey guys - useful input thanks. This will be an ongoing project after Easter and I haven't had the chance to look too far into it so far. Lets see what can be done! Cheers

[nemoc]

Perhaps sodium hydrogen carbonate (sodium bicarb) is an easier alternative for the hobbiest?

[FUGU]

Thanks nemoc - I haven't chked this out yet we'll have a look after the Easter hols. If you spot anything in the meantime please post.

Cheers

Fugu

[chew1904]

For water sterilisation we’re currently doing shock chlorination with calcium hypochlorite and sodium thiosulphate for dechlorinating, the bonus is that both the culture vessel and water are treated at the same time. Heat sterilisation is used for small volumes and starter cultures with a microwave with 1 litre on high for 7 mins and allowed to cool before adding salt.

Hi Fugu

Can you tell me how much calcium hypochlorite and sodium thiosulphate you are using per litre? I've found somewhere I can get both but I can't find any real information on the amount of thiosulphate needed to completely dechlorinate.

[FUGU]

Hi Chewi

It will be a tiny amount you need.

Approx figs:

calcium hypochlorite 0.2g per 15 litres
soidum thiosulphate 2.5g per 15 litres

Just keep the first one well away from any livestock tanks incase of accidental spills and beware that spills will bleach any fabric.

Cheers and have a nice w/e :)

Fugu

[chew1904]

Thanks Fugu. As helpful as always! :thumbsup:

[FUGU]

June 2011 update: pricing on the Isochrysis culture now in line with the other phytos

100ml: £2.80
500ml: £7.99
2.5 litres: £26.50

A lovely 90 litre brew:

http://i310.photobucket.com/albums/kk429/copeblob/really_big_icon_isoculture.png

Cheers

Fugu

[gtfcniles]

wow what diameter is that pipe! Clear plastic pipe is stupid expensive.

[FUGU]

Howdy - diameter is 12" and I've just mastered how to manage them with respect to doing production runs, standard cleaning, and then scale removal washes. Fun and games but it should be worth it. :)