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Growing phyto without a starter culture


seabass

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It's supposed to look like hammered copper; however, it's just plated with a thin layer of copper.  It is true that too much copper can act as an  algaecide.  And ideally, you wouldn't purposely use a copper container.  IDK, maybe it'll have some effect, but I think it will be alright.  Apparently, there was already some algae growing in the bath.  I'm expecting to see the culture becoming distinctly green by next weekend.

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RayWhisperer
On 8/15/2018 at 4:26 PM, A Little Blue said:

I don’t know if I want to go with under-gravel filter. Miracle Mud substrate about 1”-1.5” deep with some live rock rubble. Would love to have seagrass in the display part of the tank (commonly present in Pipefish natural surrounding) but I doubt that Seagrass will be able to thrive alongside macro algae reactor. Low flow tank, around 150gph. To maximize efficiency of the reactor and minimize flow in the display part of the tank (tank drilled and reactor used as a close loop filtration/flow). Pipefish aren’t really great swimmers and lose a lot of energy in stronger currents. Usually live in deeper waters for that reason. They feed on small crustaceans, plankton, small shrimp and copepods/amphipods, rarely accept frozen food. Light isn't important as I won’t have any LPS/SPS corals in this tank but they are visual feeders, meaning they need to see food in order to suck it in with its tinny mouth. I would like a pair of Bluestripe Pipefish that grows to max 3”. It’s possible to get a matted pair, which would be optimal. Or one male and two females. I might have missed something but that’s basically all there is to Pipefish, care needed and my plan for this setup. 

I don’t know where you are getting all of this info from. As far as MM, pod culture, or whatever else, I’ll defer to seabass and subsea. However, blue stripe are a reef pipefish, and have no problems with flow and current. They are, in fact, very strong swimmers. As are any of the flag fin pipefish. The pipefish that inhabit sea grass beds, OTOH, are weaker swimmers. Pipes like the chain, or gulf pipefish are better suited to what you are planning on.

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On 8/18/2018 at 3:15 PM, seabass said:

IDK, maybe a little more green.

081818a.jpg

 

I might add some more Miracle-Gro tomorrow.

I added 2 ml more Miracle-Gro yesterday.  Looking a bit more green today:

082018a.jpg

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So I'm almost sure that I can get an active saltwater culture from this.  However, I wondering about bacterial contaminants in the water (from birds and such).  Anybody have any thoughts about this?  Would you think that these populations would fade over time?  Any sort of filtration or sterilization would probably affect the algae too.  I've even considered using an antibiotic.  I would think that a marine salinity might kill some bacterial strains. :unsure:

 

I'm also wondering if I should try to capture algae spores indoors to limit the potential bacterial contamination.  A compromise might be letting some water sit out on our screened-in porch.  Algae spores should be able to pass through the screens, but other animals couldn't contaminate it.  Again, thoughts?  Also, is anybody generally interested in me pursuing any of these alternatives?

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Birds carry a whole host of protozoa, flukes and other parasites, so I wouldn't drink it, but the saltwater might just kill everything. The live ones would probably die, but the cysts and eggs would probably remain dormant.

If you really want to isolate the algae I'd try separating out some and moving it through several culture bottles over time, eventually you'd have a relatively pure culture.  

If you have access to microscope and can find out what algae it is, it might give you a better clue on what the best course of action would be.

 

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Um...  How much money does a phytoplankton starter culture cost?  It's the ongoing expense of repeated phyto purchases that can be saved by culturing it at home, not the initial investment in a starer culture that can benefit the hobbyist.

 

Not to say this isn't a fun experiment, but I would not put my reef at risk of contamination to save a few bucks. 

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1 hour ago, Lypto said:

Birds carry a whole host of protozoa, flukes and other parasites, so I wouldn't drink it, but the saltwater might just kill everything...  If you really want to isolate the algae I'd try separating out some and moving it through several culture bottles over time, eventually you'd have a relatively pure culture.

Yeah, I figure by the time that I continue to dilute the culture (in order to keep it growing), most of the contaminants would be gone.  I also assume that most of the harmful bacteria (and/or parasites) would starve off.  Not to mention the effect of higher salinity.  Still, it'd be nice to start with a less contaminated culture.

 

56 minutes ago, holy carp said:

Um...  How much money does a phytoplankton starter culture cost?  It's the ongoing expense of repeated phyto purchases that can be saved by culturing it at home, not the initial investment in a starer culture that can benefit the hobbyist.  Not to say this isn't a fun experiment, but I would not put my reef at risk of contamination to save a few bucks. 

Fair enough.  This has always more of an experiment (of what's possible) than an instructional article.

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After the culture darkens up a bit more, I'll probably put it under one of these cheapo digital microscopes.

image.jpeg.65f1a61f372132972f21d3ed9168651f.jpeg
I've had one for awhile, but haven't taken it out of the box yet.  I have no idea what it will show (if anything).
 
 
I strained the culture tonight with a 53 micron sieve (should have taken a pic).  There was a decent amount of green matter that was filtered out (must have been a larger algae than Chlorella).  However, the culture still has a green tint to it.
 
 
Also, I'll probably put out a bowl of RO/DI water on our porch (to catch algae spores).  I'd actually feel much better about using a culture derived from that.
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53 minutes ago, SeaFurn said:

Why not put a bowl of 1.016 saltwater out on the porch while you’re at it? 

I assume that the algae that I'd be catching is Chlorella (which is typically a freshwater algae).  So I guess I'm assuming that it would be more likely to grow in freshwater (just speculating).  Also, I'd have to top it off regularly, and I'm not sure that I plan to give it that much attention. :lol:

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This is so very interesting!

 

I’m curious why the algae, simpler in form to most, can adapt to changing salinity!  Will only further prove that FW dips are ineffective against more complex nuisance algae.

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Today:

082218a.jpg  082218b.jpg  082218c.jpg

 

Here is the culture before touching it.  I shook it, and strained it through a 53 micron plankton sieve.  Then I rinsed out the bottle, and added the culture back.

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On 8/21/2018 at 12:09 AM, Aurortpa said:

This is so very interesting!

 

I’m curious why the algae, simpler in form to most, can adapt to changing salinity!  Will only further prove that FW dips are ineffective against more complex nuisance algae.

Hydrogen peroxide, IMO, is extremely effective against all algae as a dip.  For hardy corals, I dip for 10 minutes in 10% solution of peroxide with tank water.  This dip kills everything including  pods, micro stars, bristle worms.  Slim coat protects coral.  This dip does not eliminate Aptasia.

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If I have time tomorrow, I might change half the culture with some 1.019 saltwater (and some more Miracle-Gro).  It'd be nice if I could ID what I have growing in here.  Maybe next week I'll break out the cheap digital microscope (IIRC, it might be 1000x magnification).  Although I'm not expecting much. :mellow:

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20 hours ago, Subsea said:

Hydrogen peroxide, IMO, is extremely effective against all algae as a dip.  For hardy corals, I dip for 10 minutes in 10% solution of peroxide with tank water.  This dip kills everything including  pods, micro stars, bristle worms.  Slim coat protects coral.  This dip does not eliminate Aptasia.

I have used h202 for dipping collected plants for freshwater tanks. You could see all kinds of bugs raining down from the plants, and the plants barely survive, but it sure does clean out all the nasty stuff.

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Thanks Chris!

 

Here's my culture today after straining it, and diluting it (by more than half) with 1.019 saltwater.  I also added 1ml of Miracle-Grow prior to diluting it.  Oh, and I also cleaned, bleached, and dechlorinated the bottle.  I also noted that my strainer didn't catch much of anything today (so it's probably becoming mostly phyto).

082318a.jpg

We'll have to see if the saltwater has any effect on the culture.  But it's coming along nicely.  Anybody surprised it's this easy?  Culturing phyto is so much fun.  :happydance:

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So the culture has changed a bit today.  After shaking it, the plankton sieve caught a lot more material.

082418a.jpg082418b.jpg

 

Here's what's left after filtering.  You can see how it wants to clump together.

082418c.jpg082418d.jpg

 

I broke out the digital microscope.  While pretty cool, I can't really see any more detail than what my pocket sized Canon can give me (even though it lets you get closer).

082418e.jpg

It uses the Windows 10 Camera app, so it's super simple.  I left the pic as is to let you see what I got.  IDK, maybe I can improve upon it with some practice.

 

Edit: BTW, I bought some glass slides and cover slips.  Maybe this will allow me to get even closer, while flattening the field of view (allowing for better pics).

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It's still clumping, so I filtered the culture again (so it's a little lighter). :unsure:  I don't really want to contaminate my 25 micron screen (that I occasionally use for my "real" phyto culture), so I will continue to use this 53 micron sieve on this culture.

 

I've seen other cultures do this on occasion.  I'm not sure if it's nutrient related; however, the addition of salt seems like an fairly obvious cause.

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I found this article which discusses this problem in suspended cell cultures: https://www.sigmaaldrich.com/technical-documents/articles/biology/cell-culture-troubleshooting-cell-clumping.html

 

So apparently the sudden addition of salt ruptured many of the cell walls (due to osmotic shock), releasing (sticky) free DNA, resulting in the clumping of cells.  These cell clumps would not fit through my plankton sieve.  My solution going forward will be to continue to filter the culture until the sticky cells (and DNA) are removed.

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I planned on raising the specific gravity much slower, but got impatient.  It's a good learning experience about acclimating a culture to another sg.  So I guess the remaining cells must be more compatible with higher salinites (or at least less prone to rupture due to osmotic shock).  Anyway, the first step towards adapting the culture to saltwater.

 

BTW, I strained the culture once again.  The culture is a bit lighter than when I first added the saltwater, but there was much less clumping this time.

082418f.jpg

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