DIY media for bioluminescente algae

[loïc lauréote]

Hi everybody,
Does anybody knows a DIY liquid media for growing bioluminescence algae like  Pyrocystis lunula.
I grow this algae with a standard laboratory media, but it's too expensive for massive growing.
Thank you for yours answers.
.
Thanks

Best regards

[Avery louie]

Biocurious came up with something.  It involved running water through fertilizer, and then filter sterilizing, I think.

They can also be grown in regular sterilized seawater.

--A

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[Patrik D'haeseleer]

Weird - I posted a lengthy supply, but now I only see a "This message has been deleted." Did someone accidentally push a wrong button? Can it be undeleted, so I don't have to type all that again?

Google Groups sure acts weird sometimes...

[Avery louie]

Weird, I didn't know thongs could be deleted...

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[osazuwa]

My friend and I had read somewhere that you could with miracle grow, but our initial attempt caused our dinoflagellates to die sooner than those in the normal broth they came in.
That said we didn't investigate further. Designing an experiment that investigates some of the recommended approaches would be useful.
Cheers,
Robert Osazuwa Ness

[Avery louie]

Another thing that is often understated is how long the dinos can live in f/2 media- it is certainly longer than it says on the box.  Mine had been going for 3 months w/o a media change when I dropped them on the floor... :(

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[Sebastian S. Cocioba]

Anyone have a recipe for commercial broth on hand? Maybe the paper that produced said broth might shine some light on the dino's needs. My hunch is copper content since mg has a slightly higher than average cupric quantity :)

Sebastian S Cocioba

CEO & Founder

New York Botanics, LLC

Sent via Mobile E-Mail 

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[Avery louie]

recipe: http://www-cyanosite.bio.purdue.edu/media/table/f2.html

Also, I would look into getting the cheap mass-packs from florida aqua farms/some other aquaculture supplier (I would hook you up with where I bought it from, but the bottle is in MA...I will check when I find it).

IMO, it is almost never worth it to piece together media by hand, especially if you need small amounts.  I had to make "marine broth" once, and we went nuts trying to find all kinds of crap, from strontium salts to buffers.  Each stock chemical we used was easily $90+ for the size of the bottle, but we ended up using milligrams (This is when I learned milligrams were small).  It was only feasible because we had it all on hand.

--A

[Patrik D'haeseleer]

Argh!! It deleted my post *again*. Maybe too many links?

I'll just try posting in small snippets, in case the system eats it again...

[Patrik D'haeseleer]

We've been doing lots of DIY growth experiments with P. lunula at BioCurious lately. In fact, we handed out around 250 vials with dinoflagellates for people to take home at Maker Faire a coupel months ago, along with this "Care & Feeding" link:

http:// TinyUrl.com / MyPetDinos

(To be updated with more recent results soon.)

[Patrik D'haeseleer]

The official medium (L1 or f/2) is mostly nitrate by weight, so we figured we'd give plant fertilizers a try. I like liquid Miracle-Gro, because it's easiest to find and work with, but pretty much everything we've tried works. You just get more trouble with cyanobacterial contamination at higher fertilizer concentrations, presumably because the phosphorus content is much higher than in the official medium.

We used salt water from a local aquarium store, but you should be able to avoid contamination issues by sterilizing the water, or mixing your own from filtered water plus sea salts.

[Patrik D'haeseleer]

Plant fertilizers are great if the goal is to be able to tell any ten year old how to grow their own dinos, but if you're growing them yourself in massive amounts (love to hear how massive, and why!), you're much better off buying the Micro Algae Grow Mass Pack (without silicate) from here:

http://florida-aqua-farms.com/secure/agora.cgi?product=MICRO_MACRO_NUTRIENTS

$20 plus shipping will buy you a whole kilo of dry nutrients (just add salt water), which is enough for 10,000 liters! At that rate, you'll probably pay a lot more to filter or sterilize the salt water than on the nutrient mix...

Patrik

[loïc lauréote]

Hi,

Thanks for answers,

I did a success trial with seawater 3 months ago, but i had a problem of contaminant (green algae) and the concentration didn't satisfied me.
I will  create a bioreactor with at least 100 liters and more later for a visual performance, so i need a large volume in a transparent box and the higher concentration i can.
I will try with plant fertilizer, (thanks Patrick), and try to reproduce the exact f2 medium composition for accurate reproduction.
is there a problem to provide co2 in a large volume ?

see you

Loïc

2012/8/28 Patrik D'haeseleer <pat...@gmail.com>

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[Patrik D'haeseleer]

Note that the Micro Algae Grow Mass Pack link I posted earlier *is* f/2 medium. At $20 for 10,000 liter, there's really no reason to mix your own or use plant fertilizer...

Dinoflagellates grow very slowly, doubling every 5-6 days at most, so they typically don't need much CO2. Plus they don't like constant motion. For a large volume like 100 liter, maybe get an aquarium pump that you turn on once a day or so. Heck, with some luck you might be able to find a cheap second hand 100L aquarium on craigslist, complete with light fixtures and pump. (You'll definitely also need some grow lights on a timer, if you want the dinos to "perform" on schedule.)

Patrik

[loïc lauréote]

Thanks a lot, i will build my protocole with theses recommandations.

2012/8/27 Patrik D'haeseleer <pat...@gmail.com>

Hi Loïc,

We've actually been doing a bunch of growth experiments with P. lunula at BioCurious lately. We handed out around 250 vials of dinoflagellates at the Bay Area Maker Faire a couple of months ago, along with a link to the following page of instructions:

http://tinyurl.com/MyPetDinos  (I'll be updating that one soon with our latest results)

The official growth medium (L1 or f/2) is mostly nitrate by weight, so we figured we would give plant fertilizers a try. Turns out they grow on pretty much anything we've tried, provided you get the concentration in the right neighborhood. Higher concentrations of plant fertilizer do tend to cause more problems with cyanobacterial contamination, presumably because of the higher phosphorus levels than are found in the official medium. We've been getting our salt water from a local aquarium store, but you may be able to avoid this problem by sterilizing the water, or mixing it yourself from filtered water plus sea salts.

What size of "massive" growing do you have in mind here? Plant fertilizers are great because you can tell any ten year old how to grow dinoflagellates. But if you're really planning to grow massive amounts in a more controlled environment, you may want to have a look at the "Micro Algae Grow Mass Pack" here:

http://florida-aqua-farms.com/secure/agora.cgi?cart_id=3307010.22134*Ua7qr2&product=MICRO_MACRO_NUTRIENTS

One kg of the dry nutrients only costs $20 plus shipping, and is enough to make 10,000 liters! At that level, it's going to cost you more to filter or sterilize that much salt water...

On Monday, August 27, 2012 2:16:11 AM UTC-7, Loïc wrote:

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[loïc lauréote]

Hi,
i have my nutrients from florida-aqua-farm, but i have one question, im not sure of my product composition, do i have to put sea salt in this aqua farm media   ?

thx,

2012/8/30 loïc lauréote <loic.l...@gmail.com>

[Patrik D'haeseleer]

Hi Loïc,

Did you wind up getting the "mass pack"? In th ecase of the liquid nutrients, you add just 10 drops per liter of salt water. I assume the mass pack works the same: just add salt water.

You can buy salt water at any aquarium store - around a dollar per gallon around here. If you really need huge volumes, I guess you could filter seawater as well, but it's hard to compete with the convenience and cost of aquarium store salt water.

Word of warning: we've had a lot more trouble with cyanobacterial contamination from one particular aquariums store. Might want to look around and experiment a little.

Patrik

[loïc lauréote]

i have the mass pack, i have to add salt.
thx

2012/10/12, Patrik D'haeseleer <pat...@gmail.com>:
> Hi Loïc,
>

> Did you wind up getting the "mass pack"? In th ecase of the liquid
> nutrients, you add just 10 drops per liter of salt water. I assume the mass
>
> pack works the same: just add salt water.
>
> You can buy salt water at any aquarium store - around a dollar per gallon
> around here. If you really need huge volumes, I guess you could filter
> seawater as well, but it's hard to compete with the convenience and cost of
>
> aquarium store salt water.
>
> Word of warning: we've had a lot more trouble with cyanobacterial
> contamination from one particular aquariums store. Might want to look
> around and experiment a little.
>
> Patrik
>
> On Tuesday, October 9, 2012 2:44:18 AM UTC-7, Loïc wrote:
>>
>> Hi,
>> i have my nutrients from florida-aqua-farm, but i have one question, im
>> not sure of my product composition, do i have to put sea salt in this aqua
>>
>> farm media ?
>>
>> thx,
>>

>> 2012/8/30 loïc lauréote <loic.l...@gmail.com <javascript:>>

>>
>>> Thanks a lot, i will build my protocole with theses recommandations.
>>>
>>>
>>>

>>> 2012/8/27 Patrik D'haeseleer <pat...@gmail.com <javascript:>>

>>>
>>>> Hi Loïc,
>>>>
>>>> We've actually been doing a bunch of growth experiments with P. lunula
>>>> at BioCurious lately. We handed out around 250 vials of dinoflagellates
>>>> at
>>>> the Bay Area Maker Faire a couple of months ago, along with a link to
>>>> the
>>>> following page of instructions:
>>>>
>>>> http://tinyurl.com/MyPetDinos (I'll be updating that one soon with our
>>>>
>>>> latest results)
>>>>

>>>> The official growth medium (L1 <https://ncma.bigelow.org/node/83> or
>>>> f/2<https://ncma.bigelow.org/node/79>)

>>>>> like *Pyrocystis lunula.
>>>>> *I grow this algae with a standard laboratory media*, but *it's too
>>>>> expensive for massive growing*.
>>>>> *Thank you for yours answer*s.
>>>>> .
>>>>> *Thanks
>>>>>
>>>>> Best regards*
>>>>> *

>>>>
>>>> --
>>>> You received this message because you are subscribed to the Google
>>>> Groups "DIYbio" group.
>>>> To post to this group, send email to

>>>> diy...@googlegroups.com<javascript:>
>>>> .

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>>>> diybio+un...@googlegroups.com <javascript:>.

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>>>> https://groups.google.com/d/msg/diybio/-/Ar-bQUL_bH8J.
>>>>
>>>> For more options, visit https://groups.google.com/groups/opt_out.
>>>>
>>>>
>>>>
>>>
>>>
>>
>
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[Meow-Ludo]

Hi Patrik,

I live in Australia and was wondering if there is a protocol for extracting them from seawater? I have learned a little bit about them at uni and am keen to play with them. I live about 30m away from the ocean so supplying them with fresh nutrients is quite easy.

Are there restrictions on getting them sent internationally?

Cheers,

Meow

[Patrik D'haeseleer]

Hi Meow,

The guys we get our dinos from are actually located here in the San Francisco area. But it does look like they can ship oversees, based on their FAQ page: http://empco.org/edu/faq

Many dinoflagellates produce toxins, so that's something you might want to take into account before culturing some from seawater.

Not sure how to isolate them yourself. You might want to have a look in some of the older papers to see how they did it. You may be able to grow them on fairly sparse medium, and kill of the cyanobacteria using antibacterials

Patrik

[Mega [Andreas Sturm]]

Does anyone know whether you can adapt the dinoflagellates (P. Fusiformis) to lower salt concentration / freshwater? 

[Brian Degger]

Seawater + garden plant fertilizer?

Mega why do you want to grow them in less salty water. Can get contamination with potentially bad bacterias.

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[Patrik D'haeseleer]

On Wednesday, July 31, 2013 1:54:43 AM UTC-7, Mega [Andreas Sturm] wrote:

Does anyone know whether you can adapt the dinoflagellates (P. Fusiformis) to lower salt concentration / freshwater? 

I doubt anyone has tried yet. They do grow very slowly as is though, so they'll take a longtime to evolve to adapt to the lower salt concentration. And even if you can grow them in lower salt, they may grow sooo slow it might not be worth your time.

I'd love to be able to find some way to grow them at much higher concentrations though - that might be feasible. I suspect they're producing some sort of byproduct that limits their own growth. We might be able to figure out what that limiting factor is - or at least narrow down the options - by centrifuging down and fractionating the supernatant.

Another option might be to test a range of marine bacteria in a co-culture with the dinoflagellates, in case one of them can metabolize whatever the dinos are excreting, and allow them to reach higher concentrations. Plenty of known examples of symbiotic relationships between bacteria and dinoflagellates or other algae...

Patrik

[Andreas Sturm]

Would be nice to grow them in a freshwater pond of a friend. If not possible, we have to put salt in that pond :D

As they are non-toxic, I guess they would be harmless, yet very cool when your pond glows at night ;)

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[Ravasz]

How about cloning glow genes into some freshwater algae? :P My guess is you'd do better in the long run.

[Cathal Garvey]

Probably because he can't then legally grow them outdoors in a pond
where it'd look cool.

I wonder are there freshwater relatives of those bugs in the wild
though, and whether certain salt concentrations make them receptive to
DNA. Is it really your fault if some naturally competent bacteria were
to encounter some dead cousins under the right natural conditions? :P
(Don't do this)

Yet more of the illogic surrounding current GMO regulations in Europe,
which assumes that outside of human intervention, genetic modification
never occurs.

On Thu, 1 Aug 2013 02:23:44 -0700 (PDT)
Ravasz <ravasz...@gmail.com> wrote:

> How about cloning glow genes into some freshwater algae? :P My guess
> is you'd do better in the long run.
>
> On Wednesday, 31 July 2013 11:20:10 UTC+2, DrBrian wrote:
> >
> > Seawater + garden plant fertilizer?
> >
> > Mega why do you want to grow them in less salty water. Can get
> > contamination with potentially bad bacterias.
> > On 27 Aug 2012 12:31, "loïc lauréote" <loic.l...@gmail.com

> > <javascript:>> wrote:
> >
> >> Hi everybody,
> >> Does anybody knows a DIY liquid media for growing bioluminescence

> >> algae like *Pyrocystis lunula.
> >> *I grow this algae with a standard laboratory media*, but *it's
> >> too expensive for massive growing*.
> >> *Thank you for yours answer*s.
> >> .
> >> *Thanks
> >>
> >> Best regards*
> >> *
> >>

> >> --
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> >> Groups "DIYbio" group.
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> >> diy...@googlegroups.com<javascript:> .

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[Andreas Sturm]

> Probably because he can't then legally grow them outdoors in a pond

> where it'd look cool.

Exactly... 

> I wonder are there freshwater relatives of those bugs in the wild

> though, and whether certain salt concentrations make them receptive to

> DNA.

Haha, that would be fine... 

But they're eukaryotes, so only (legal) means to achieve this would be cross-breeding with freshwater algae? There are at least four genes involved, one that produces the light organelles (scintillons), luciferase, luciferin (or a few genes), a luciferin binding protein. 
Perhaps also a circadian rythm gene which activates the glow. And something that drops the pH in the scintillons, (IIRC, they need pH 6)

>Yet more of the illogic surrounding current GMO regulations in Europe,

which assumes that outside of human intervention, genetic modification

never occurs.

You say it... Crazy, gotta move to US^^

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[Andreas Sturm]

>Yet more of the illogic surrounding current GMO regulations in Europe,

which assumes that outside of human intervention, genetic modification

never occurs.

Isn't this to some degree a form of Creationism, which they say Americans believe in it? 

They believe God created everything, no evolution occured, no genes are trandsferred between species, ... You shouldn't mess with the genes of organism. 

Why isn't it the other way round, that America is against GMO and Europe not? 

[Nathan McCorkle]

I'm not up to date on what messages religious groups are pushing these days, but i'be heard Catholics have embraced evolution as a god-based mechanism. I guess its good to have some more people hopping on board whether they say physics is behind it or god.

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