Growing phyto without a starter culture

[Christopher Marks]

This thread had me looking for mad scientist gifs, but I got distracted by these algae ones instead 😄 

 

 

 

[seabass]

50 minutes ago, Lawnman said:

One of my customers has some green phyto looking stuff in his fountain lol. I was going to snap a picture today. 

Haha, you should have.   It could possibly be the same species.  Spores seem to make it to just about any body of water.

 

Maybe in a week or so, I'll try to culture the water from our screened-in porch.  It receives no real sun to speak of, so it won't have any tint to it at all.  However, I'm still hoping that algae spores get captured in the water.  Should make for a less contaminated culture (although after awhile, I bet this culture would be fine too).

 

 

38 minutes ago, Christopher Marks said:

This thread had me looking for mad scientist gifs, but I got distracted by these algae ones instead 😄 

  That pod! 

[seabass]

Just to continue documenting the progress.

   

1) before shaking,  2) strained cells,  3) back in the bottle

 

I think the culture will be fine.  For reference, I tested the sg this morning (it was 1.010).  More nutrients tomorrow.

[seabass]

Considering the algae which died due to osmotic shock, I'm reminded of one of my favorite quotes from Clownfishes, "Do not be discouraged if you have 1, 2, or 200 rotifer crashes. Just start the culture again with the cyst-containing sediment from the bottom of the crashed culture and call it genetic honing. You’ll eventually get some super rotifers that thrive on the quality of care you give them. Rotifers procreate through about 80 generations a year, with each new generation “selected” by the stresses that their parents endured. By now, I’m fairly certain that my cultures are a hardy strain—many generations ago, I killed off those that could not take my particular brand of TLC."

- Wilkerson, Joyce D.. Clownfishes (Kindle Locations 4210-4214). Microcosm Ltd.. Kindle Edition.

 

So I just consider it genetically honing my phyto culture.

[Subsea]

4 hours ago, seabass said:

Considering the algae which died due to osmotic shock, I'm reminded of one of my favorite quotes from Clownfishes, "Do not be discouraged if you have 1, 2, or 200 rotifer crashes. Just start the culture again with the cyst-containing sediment from the bottom of the crashed culture and call it genetic honing. You’ll eventually get some super rotifers that thrive on the quality of care you give them. Rotifers procreate through about 80 generations a year, with each new generation “selected” by the stresses that their parents endured. By now, I’m fairly certain that my cultures are a hardy strain—many generations ago, I killed off those that could not take my particular brand of TLC."

- Wilkerson, Joyce D.. Clownfishes (Kindle Locations 4210-4214). Microcosm Ltd.. Kindle Edition.

 

So I just consider it genetically honing my phyto culture.

Exactly my sentiments as a “Laissez Faire” reefkeeper.

Laissez les bonne temps roulee,

Patrick

[Aurortpa]

On 8/20/2018 at 2:42 PM, seabass said:

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.

 

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?

I work in an animal sanctuary...some avian bacteria have the ability to go dormant for years like psittacosis--a form of chlamydia. However, I would think that the osmotic change would destroy them, as these observations were made in hosts but honestly I am not absolutely certain.  My expertise is in behavior and husbandry, not pathology.  

[seabass]

I didn't shake the culture today; just strained it, added 2ml of Miracle-Gro, and cleaned the bottle.

 

1) Prior to straining 2) After straining

[paneubert]

Your silence is killing me.....

[seabass]

Today (08/30/18):

 

 

[seabass]

I think that next week I'll dump this culture, and try to culture algae from the water that has been sitting on my porch (phase 3).  Again, my intention with the bird bath phyto wasn't necessarily to use this culture in my systems.  However, it was an interesting exercise nonetheless.  I'll post a final update on this phase on Sunday or Monday.  Maybe I'll add another 2ml of Miracle-Gro as a final boost.

[Lypto]

It seems like its working pretty well! Curious to see if it holds up!

[Ladytank]

After a month of research, this it what I found. You should be using f2 fertilizer. Its for planted tanks and you get it at your lfs. Miracle grow can crash your tank. That seems pretty expensive to me if your on a tight budget to begin with. Just my thought. Cheaper to buy a bottle and cultivate it from there. For about 30 you can have a sustainable solution.

[seabass]

13 hours ago, Ladytank said:

After a month of research, this it what I found. You should be using f2 fertilizer.

You are correct of course; that is the recommended fertilizer to use.  I mostly use Guillard's f/2 formula (Micro Algae Grow, from Florida Aqua Farms) to feed my Nannochloropsis oculata culture:

 

But I actually own, and have used, both (Miracle-Gro and f/2) for culturing rotifers, and even sometimes for use in my tanks.  The cost difference seems fairly insignificant.  I was intentionally trying to keep these trials low tech, and able to be replicated without ordering anything online (since not everyone has a LFS which stocks f/2).  We could go a step further and ask, why not just pick up some live phyto at your LFS; but what fun is that. 

 

In that same vein, I used a coffee filter instead of a plankton screen when collecting the bird bath culture.  However, granted, I started using a screen during the culturing process, purely as a convenience.

 

13 hours ago, Ladytank said:

Miracle grow can crash your tank.

I'd be interested in what you found, because I've never seen that proven out.  Sure, there's speculation (based on it containing things like copper) that it might be harmful to reef systems.  However, copper is also included in Guillard's f/2 (which uses chelated copper sulfate).  In fact, many reef supplements contain copper (even our salt mixes).

 

For anecdotal evidence, I'll tell you that if Miracle-Gro were that toxic to marine inverts, then I wouldn't think that marine rotifer cultures would be sustainable in phyto cultures fed with it (since the rotifers are directly exposed to the phyto culture, as opposed to the few milliliters dosed into our tanks).  However, rotifers reproduce into the millions when using phyto fed with nothing but Miracle-Gro.

 

That doesn't mean that I'm recommending the use of Miracle-Gro over f/2.  It simply means that maybe it's not as bad as some might make it out to be.  Again, this has been more of a working thought experiment, and an exercise in starting a viable phyto culture from not much of anything.  Extra care is probably warranted for cultures that will be used in our systems.

[Subsea]

If we are what we eat, then is it worthwhile to indulge with quality.  I use f/2 on phyto & macro because I think it is worthwhile, yet when it comes to organic price tags in the grocery store, I go with economy.  Both work.

 

Both copper & zinc show up in parts per billion in dry analysis of macro that I grow.  At the time, I was focused on food grade Red Ogo and wanted to present dry analysis to executive chefs in the Austin area.  I was most surprised with the high level of sulfur in dry analysis at almost 5% or 50,000 ppm.  Because of the bitter sulfur test, I could not market this as a food grade product.

 

Test of Trinity Aquifer water showed zero Fe, N, P, Mn, Cu, Zn with detection limits at 0.01 ppm.  SO4 at 190 ppm.  Dry analysis for Gracilaria Parvispora showed copper at 7 ppm, Zn at 140 ppm.  Both of these heavy metals are required as a catalyst  in metabolic processes.

[seabass]

In addition to what @Subsea mentioned - f/2 contains vitamins: B12, B1, and Biotin.  Phyto produced with it, is more nutritious than phyto grown with Miracle-Gro (that's reason enough to choose it).  Guillard's f/2 formula was formulated to meet the specific needs of algae (with nutrients, trace elements, and vitamins).  However, IME, Miracle-Gro seems to grow phyto just about as easily.

[seabass]

 

So here's the culture today (08/31/18).  Before shaking, the cells still seem to group together.  After shaking, the culture looks much more uniform.

 

So I did one more thing today.  I added 3ml of Kent Essential Elements.  I typically add this (in a smaller amount) along with Miracle-Gro to provide additional trace elements.  I have noticed that the algae is lighter (more ecto slime green) than my Nannochloropsis oculata culture.  I'm not completely sure if that's just the natural color of this species, or if it's nutrient related.  So I wanted to see if the addition of Kent's Essential Elements would make any difference.

[seabass]

I didn't really notice the clumping as much until after I added the saltwater.  I suppose it's possible that even though the culture is obviously producing new algae in this higher specific gravity, that the some of the cells may still rupture more easily (causing the grouping/clumping).  However, I'm just speculating and thinking of possible reasons why it might want to do this.

[paneubert]

I don't want to hijack your experiment thread, but what do your experienced eyes think of the darker clumps on the walls that I had in one of my batches?  Contamination?  It came off really easy when I rinsed them out.

 

 

 

[seabass]

6 minutes ago, paneubert said:

... what do your experienced eyes think of the darker clumps on the walls that I had in one of my batches?  Contamination?  It came off really easy when I rinsed them out.

That's totally normal.  I use a bottle brush to clean that off whenever I harvest the phyto.  Shaking the culture often releases many of these cells, although sometimes it comes off in larger chunks (which can be removed with a plankton sieve or even a brine shrimp net).

 

I've read where people have used bleach or even acid to clean the bottles.  However, a bottle brush has always been sufficient for me.  If you don't clean it off, it will block some of the light.

[paneubert]

21 minutes ago, seabass said:

 

I've read where people have used bleach or even acid to clean the bottles.  However, a bottle brush has always been sufficient for me.  If you don't clean it off, it will block some of the light.

In yet another example of my saltwater life crossing over to my homebrew/cider making life, I have a crap ton of StarSan concentrate laying around.  I switched to new style jugs for my next phyto batch, otherwise I would have probably soaked these ones in StarSan for a bit.  Killing algae is not technically the labeled use for StarSan but it sure kills yeast cells efficiently, is specifically labeled to work well on "soiled" surfaces, and is actually food safe.  You can technically drink it when it is diluted.....but it is deadly to smaller lifeforms.  Maybe I should start a StarSan revolution in the saltwater community.  Hahaha.

[Lawnman]

Got a picture of that fountain 🙂 

[seabass]

Wow, talk about greenwater.  That's crazy.

[seabass]

I'm pretty happy with how things worked out with this culture (which I started from water that I collected from our bird bath).  It started off as a freshwater culture that looked completely clear.  But today, it was a brackish culture (1.010 sg) that was distinctly green.  I'm kind of sad about it; but seeing that this was just an experiment, I discarded the culture today.

 

09/03/18:

 

The culture didn't look that bad, even before shaking.  And unlike the last few times that I've strained the culture, this time only a little material was filtered out.  This is much more representative of a typical phyto culture.

 

 

Here is the culture after shaking.  I also broke out the slides and digital microscope.

 

Here are the pics from the cheap digital scope that I have.  For reference, the silver lines are 5mm apart.

 

You can't see much, but I thought I'd take a couple of pics before I ditched the culture.

[Christopher Marks]

This has been really interesting to follow along with! What a neat outcome, all from a bird bath!

[seabass]

Phase 3 of 4 (capturing algae spores, porch phyto)
In Phase One, I was trying to replicate the results of Joyce D. Wilkerson (author of Clownfishes); where she observed that a percentage of the jugs of distilled water that she left out in the sun, eventually became tinted green with algae growth.  However, I was unsuccessful at duplicating similar growth in various types of water stored outside in bottles (leading me to believe that her experience may have been a one off).

 

In Phase Two, I was fairly certain that I could start an algae culture from a freshwater pool, which already had some traces of algae in it.  IMO, this phased proved to be a success.

 

Now in Phase Three, I hope to capture algae spore cells with a bowl of water that was placed on my screened-in porch for a couple of weeks.  While no evidence of algae currently exists, I will use the same culturing process as I did in Phase 2.  Hopefully, captured spores will provide the basis for a new freshwater phyto culture, that I will eventually acclimate to a higher specific gravity.

 

     

This is an old, but clean, food storage container that was previously stained with tomato sauce (the water isn't as discolored as it appears).  Here it is after being strained through a paper towel.

 

I then added 4ml of Micro Algae Grow from Florida Aqua Farms (Guillard’s f/2 fertilizer) to the porch water.  I filled the rest of the bottle with RO/DI and will light it with a standard household 75W equivalent LED bulb (daylight spectrum) for 24 hours a day.  As you can see, there are no visible signs of algae present at this time.  However, I'm still cautiously optimistic that this effort will be successful.

 

Note:  I disinfected (with bleach and then with hydrogen peroxide) the equipment that was used to culture my bird bath phyto, as I will be reusing it for this phase of the experiment.  While there is some chance of cross contamination, I tried to limit that risk.