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My Battle with Dinos


obrien.david.j

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Harvest #28 complete (Jan28th)

  • Continuing to add mid-grow fertilizer kicker.   (2nd dose, after about 1wk) Harvest is back to dark-dark grow of the past

Thought you might like to see the setup, ready to harvest.  Empty bottles ready to go, towel down to catch all drippings, polypropylene beakers, fertilizer, and most importantly isopropyl alcohol to keep things "sterile."   (not really sterile, but to wipe everything down with and keep contamination to a minimum) 

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The harvest results

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Great work on the thread + phyto + pods. Just curious about doing the fertilizer boost to thicken your harvest. Did you ever test for excess fertilizer? 

I had to alter my carbon ratio to light duration to avoid a boom/bust cycle of adding f/2. Nanno oculata is a great strain but long term continuous yielded headaches for me that led to me doing batch cultures for ease of maintenence. I do continuous cultures on pretty much everything else minus those that require silicates (batch gives me greater controll of excess nutrients). 

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1 hour ago, Eatfrenchfries said:

Great work on the thread + phyto + pods. Just curious about doing the fertilizer boost to thicken your harvest. Did you ever test for excess fertilizer? 

I had to alter my carbon ratio to light duration to avoid a boom/bust cycle of adding f/2. Nanno oculata is a great strain but long term continuous yielded headaches for me that led to me doing batch cultures for ease of maintenence. I do continuous cultures on pretty much everything else minus those that require silicates (batch gives me greater controll of excess nutrients). 

I've always wondered how I test for excess fertilizer.   Can you tell me how/give me a procedure?   I've got Hana nitrate and phosphate test kits.

All my phyto is Nanno, and I'm only doing batch cultures.    My idea being Fert getting fully used is the instructions that came with this system were 2ml of A, 2ml of B Fert, for 1gallon grow container - harvest after 7-10days.   When I did that, cultures were weak/pale/not as dense.   I'm now doing mid cycle kicker of Fert, and harvesting at 15-20days - super dense/dark.   I am restarting each gallon with 300ml of the last batch.

 

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For the longest time I used a 3D printed dremel attachment to spin cultures for testing the water. Just got myself a used centrifuge for cheap on FB that is nice but no real need for it. I spin the culture till it separates and then I test the clear portion. Works like a charm. Just make sure it is the clear portion and not the phyto itself. The color will ruin your test. Even better would be sending that water to be tested to see what residuals your phyto didn't consume. 

 

Poseidon ferts come separated to prolong shelf life since they come premixed. Mixing your own removes precipitation issues and guarantees ratios for consumption. Whenever I start my phyto culture I use a 1 (culture) : 5 (medium) to start up. Certain prolific ones I start at 1:3. 

 

Usually for Nanno since it is nonmotile and prone to failing thru contamination I use a denser culture to start it. They thrive in density. I recommend 1:3 or 1:1 even. I prefer tetra in bulk but I keep nanno going because it is useful. 

Your cells would be dying right around the time you re-up them on fertilizer. They're using that fertilizer + decayed cells (carbon) to fuel further growth. Phyto thrives on density. Dilution leads to problems (except spriulna + synechococcus, all bloom no bust). 

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18 hours ago, Eatfrenchfries said:

For the longest time I used a 3D printed dremel attachment to spin cultures for testing the water. Just got myself a used centrifuge for cheap on FB that is nice but no real need for it. I spin the culture till it separates and then I test the clear portion. Works like a charm. Just make sure it is the clear portion and not the phyto itself. The color will ruin your test. Even better would be sending that water to be tested to see what residuals your phyto didn't consume. 

Spin out the phyto, test the clear water.   Am I really looking for Zero?     How many days after using "all" the fert, does the phyto culture crash.

I add 32oz (1/4gallon) phyto a day to about a 400gallon setup.   So whatever I measure in the Phyto water, divide by 1600 - and that's how much fert I'm inadvertently adding to the tank.

18 hours ago, Eatfrenchfries said:

Poseidon ferts come separated to prolong shelf life since they come premixed. Mixing your own removes precipitation issues and guarantees ratios for consumption. Whenever I start my phyto culture I use a 1 (culture) : 5 (medium) to start up. Certain prolific ones I start at 1:3. 

Your cells would be dying right around the time you re-up them on fertilizer. They're using that fertilizer + decayed cells (carbon) to fuel further growth. Phyto thrives on density. Dilution leads to problems (except spriulna + synechococcus, all bloom no bust). 

How do I know the concentration of Poseidon Fert?    It's their mix, and their recommendation of 2mlA+2mlB per ~gallon jar (which is really 3L of water - I measure when harvesting).    3L:4ml   ?

Or, is the math really about amount of starter culture to fert?    Poseidon recommends 250ml of culture w/ 2mlA+2mlB fert.   250ml:4ml  ?

Shouldn't the amount of fert be related to the density of culture desired in the final total volume of water.  Assuming a batch grow.

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When test for fertilizer you're not looking for zero. Just looking to see that you are indeed using majority of the fertilizer. I use the redfield ratio as frame of reference to see what is being residually added to my tank. Key factor for me since I keep various macroalgae. No real guess how many days after fert the culture will crash. Could be a day or hours depending on light/heat. More realistic would be your culture being on the decline and just trying to harvest before all the goodies are gone.  

Not sure if Poseidon fert is proprietary but I know they all follow the Guillard recipe with slight modifications based on personal findings. From the measurements you're telling me it sounds like the fertilizer was just diluted a bit more. I think Fritz does it the same. 

The amount of fert is related to the density of culture desired in the final total volume of water. Your culture age(density of old to young, rest or active) is also a factor. These are all things to fine tune your harvests but if you're using Poseidon systems then I would follow their guide. The owner did a great job and offers great support for a streamlined product. Their guarantee is pretty awesome too.  Maybe see if the fresh batch of fert will keep you from doing an extra squirt into your cultures. Something I took the time to dial in because managing 17+ separate cultures isn't something that leaves a lot of room for error. 

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@Eatfrenchfries

Question for you, Can you think why cultures may settle out/go light - right after adding mid cycle fert kicker?

I've noticed this the last two times I gave the 2nd dose.    1-2 days afterwards, the cultures jars look Much clearer - but bounce back to even darker than before in another 1-2 days.   (meaning, they're fine at harvest)

Added 2nd round of Fert last night.  One culture jar is clear this morning, as compared to it's peers left and right.  I expect them to temporary also go clear today/tomorrow.  And then everyone recover just fine.

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The old has to make way for the new. Dead cells will clump to form detritus which is helpful in moderation. It feeds bacteria that produce co2 and recycles the dead cell walls to an extent. Too much and the bacteria will overtake your culture leading way to competition. Your nanno will bounce back but it always runs the risk of coming back mixed with something else. Cultures that go clear and back to color make me wary since phyto isn't that fast reproducing (greens at least, a golden might with appropriate carbon, usually othe single celled bacterias can do what you are describing). Tinted like tea is fine. Yellowish tint is fine. But clear or cloudy gives me reason to pause. Especially if it goes back to green within the next day. Only  my cyano strains grow that fast. Usually a day to three to get a new batch. 

 

Reviewing your journey I would say your nanno culture could probably run even denser which would help your initial harvests. Itd keep other things from outcompeting your phyto. Retiring phyto and giving cultures breaks is a good idea too. Microscope would help too to make sure your culture is still relatively pure. Lots of peoples cultures get lost over time but still produce similiar product results for feeding pods. Not so much for tank dosing. 

*amscope is my favorite brand for microscope in terms of quality and price. 

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8 hours ago, Eatfrenchfries said:

The old has to make way for the new. Dead cells will clump to form detritus which is helpful in moderation. It feeds bacteria that produce co2 and recycles the dead cell walls to an extent. Too much and the bacteria will overtake your culture leading way to competition. Your nanno will bounce back but it always runs the risk of coming back mixed with something else. Cultures that go clear and back to color make me wary since phyto isn't that fast reproducing (greens at least, a golden might with appropriate carbon, usually othe single celled bacterias can do what you are describing). Tinted like tea is fine. Yellowish tint is fine. But clear or cloudy gives me reason to pause. Especially if it goes back to green within the next day. Only  my cyano strains grow that fast. Usually a day to three to get a new batch. 

 

Reviewing your journey I would say your nanno culture could probably run even denser which would help your initial harvests. Itd keep other things from outcompeting your phyto. Retiring phyto and giving cultures breaks is a good idea too. Microscope would help too to make sure your culture is still relatively pure. Lots of peoples cultures get lost over time but still produce similiar product results for feeding pods. Not so much for tank dosing. 

*amscope is my favorite brand for microscope in terms of quality and price. 

I've got a Swift SW350T 40X-2500X Magnification, Siedentopf Head.   I'll scope out the cultures and see what I see.

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Nanno should be round. Anything oblong would a bacteria. Nothing should be moving. Your microscope is more than enough to do the deed. 

The only other thing I can think of is your 2nd F/2 boost is pushing the culture past the exponential growth curve and into another decline and then the next curve. 

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You are nailing what I need for scoping.  Look for round, and no other shape.  And no movement.  Reference photo I found below.

Exponential Growth curves!  Now you're speaking my language.  Another round of google searches, and I can't (easily) find growth curve studies, against fertilizer additions.  But I did find a nice simple study against initial density, light intensity, salinity, and fertilizer types.

https://www.researchgate.net/publication/353922072_Effects_of_initial_density_nutrient_medium_salinity_and_light_intensity_on_the_growth_of_microalgae_Nannochloropsis_oculata

I found this pict off etsy (it's what came up from a google search)

Nannochloropsis Salina algae phytoplanktonlive image 1

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Research gate is my go to. Chances are if there's something you need access to. Just reach out for permission. Hydrospace LLC pointed me towards some good literature for my projects too. Dinkins, Reef Nutrition, Tommy's, Jay's  pointed me towards other directions that broaden my scope but didn't narrow my field enough either. They would rather show you the direction to take then the way itself.  

 

I'll tell you that added N will lead to a higher curve values than limited P. You're also best off keeping a journal for reference. I keep a log for each strain in my collection and another for those in production. Majority of people I know haven't dabbled in them and those that do guard trade secrets closely because they paid for research grade samples. They're not cheap or easy to work with. $125 - $200 for a 5mL - 100 mL sample. Possibly at the next meeting I can bring some of my journals + notes for you to look over. 

 

If growth curves are your language then you should look into a dissolved oxygen meter and pH meter. It'll help you calculate the curve. I have them both if you need. 

 

Higher pH means bigger phyto boom. Lower pH means a phyto decline. Dissolved oxygen will vary between night and day. It is important to see how much the o2 the phyto is pulling night vs day. Obviously you want it pulling during the day. If it's pulling more at night then it is contributing to your busts. 

 

Picture is of nanno at a lower magnification because I was looking for zooplankton. Obviously parvocalanus don't eat nanno but I heard they eat Thalassiosira in addition to their staple Isochrysis. Had to give it a test though to see if a diverse blend would be better than two or three big fatty motiles. 

 

Screenshot_20230204_094210_Gallery.jpg

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Still trying to find the sweet spot for my Nanno phyto as well.  I know Poseidon's instructions say harvest after 7-10 days, but as David mentioned, it's just not as dark as I'd prefer.  However, anything after 12 days or so is pretty much a coin flip, on whether or not the culture crashes and starts going clear (it's happened when I've added fertilizer after a week, and without adding anything). 

I've got 3 jugs proceeding nicely at 9 days now, so I'm thinking about harvesting 2 of them after 10 days, then try to get the 3rd to 14 days. 

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1 hour ago, Eatfrenchfries said:

Research gate is my go to. Chances are if there's something you need access to. Just reach out for permission. Hydrospace LLC pointed me towards some good literature for my projects too. Dinkins, Reef Nutrition, Tommy's, Jay's  pointed me towards other directions that broaden my scope but didn't narrow my field enough either. They would rather show you the direction to take then the way itself.  

 

I'll tell you that added N will lead to a higher curve values than limited P. You're also best off keeping a journal for reference. I keep a log for each strain in my collection and another for those in production. Majority of people I know haven't dabbled in them and those that do guard trade secrets closely because they paid for research grade samples. They're not cheap or easy to work with. $125 - $200 for a 5mL - 100 mL sample. Possibly at the next meeting I can bring some of my journals + notes for you to look over. 

 

If growth curves are your language then you should look into a dissolved oxygen meter and pH meter. It'll help you calculate the curve. I have them both if you need. 

 

Higher pH means bigger phyto boom. Lower pH means a phyto decline. Dissolved oxygen will vary between night and day. It is important to see how much the o2 the phyto is pulling night vs day. Obviously you want it pulling during the day. If it's pulling more at night then it is contributing to your busts. 

 

 

I gave away my motivation for growing phyto and pods on the very first post of this thread.  Die Dino's, Die.

My research pointed to Add Phyto (didn't say what strain), and Tisbe Pods had been seen to eat certain dino strains.   Ala Poseidon, Nano and Tisbe pods.    A good presentation at MACNA'22 suggested that Tet. much more nutritious.   Tell me about the Degree-of-Difficulty in growing it, if I wanted to switch a jar over?

Thanks for the example scope pict.

 

 

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Tetraselmis is much more nutritious. You can grow it in your poseidon system the same way. You'd just have to learn it's color palette and possibly give it a little more rest to light. No change in degree of difficulty other then your sterilization techniques to avoid cross contamination are paramount to keeping different strains. Airborne transmission happens. Tetraselmis will absolutely outcompete your Nannochloropsis. Motile vs nonmotile. 

 

Your tisbe pods may eat the dino but will die not long after, keeping it still in your tank. Dinos respond to predators. They are known to wipe out copepod, amphipod, isopod populations. Filter feeders are shown to be much better consumers of dinos. I culture a couple for that exact reason. Dosing dinos will not beat dinos, you'll just have a diversity of dinos. The issue would be figuring out how to suspend the dinos from the sand (diamond goby, pistol shrimp, etc). Your filter feeders (bivalves are awesome) would consume dinos from your system. That in addition to dosing phyto may tip the single cell organism balance in your tank away from the dinos. 

 

I keep macroalgae themed tanks. Dinos are pretty much damning since they'll melt away the tissue and disintegrate the macros. Nutrient imbalances coupled by not enough biodiversity were my biggest hurdles. Dinos are ever present in your reef, the key is to keep them at low numbers. 

 

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1 hour ago, Eatfrenchfries said:

Tetraselmis is much more nutritious. You can grow it in your poseidon system the same way. You'd just have to learn it's color palette and possibly give it a little more rest to light. No change in degree of difficulty other then your sterilization techniques to avoid cross contamination are paramount to keeping different strains. Airborne transmission happens. Tetraselmis will absolutely outcompete your Nannochloropsis. Motile vs nonmotile. 

 

Your tisbe pods may eat the dino but will die not long after, keeping it still in your tank. Dinos respond to predators. They are known to wipe out copepod, amphipod, isopod populations. Filter feeders are shown to be much better consumers of dinos. I culture a couple for that exact reason. Dosing dinos will not beat dinos, you'll just have a diversity of dinos. The issue would be figuring out how to suspend the dinos from the sand (diamond goby, pistol shrimp, etc). Your filter feeders (bivalves are awesome) would consume dinos from your system. That in addition to dosing phyto may tip the single cell organism balance in your tank away from the dinos. 

 

I keep macroalgae themed tanks. Dinos are pretty much damning since they'll melt away the tissue and disintegrate the macros. Nutrient imbalances coupled by not enough biodiversity were my biggest hurdles. Dinos are ever present in your reef, the key is to keep them at low numbers. 

 

We're out of town this weekend.  Will update after getting home tomorrow and breaking out the microscope.

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I’m late to this party but thought I’d share that I had knocked back Dino’s with UV and regular turkey basting and sand mixing to get it in the water column. Was way better but still had some collecting on the rocks and sand everyday (which generally wouldn’t significantly regenerate until the following day vs an hour like before.

Was tired of having the eyesore of my UV running out of the display so I got a big bottle of an isochrysis culture and started dosing daily. The Dino’s completely disappeared after a week. Hesitant to attribute causation, but it did seem to line up to suggest a relationship. I didn’t verify but the dinos seemed to be Ostreopsis or possibly Coolia. 
 

If the Phyto was only helping by boosting pod populations, I wouldn’t have expected to see such a big change in only a week.

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6 hours ago, Webfoot said:

I’m late to this party but thought I’d share that I had knocked back Dino’s with UV and regular turkey basting and sand mixing to get it in the water column. Was way better but still had some collecting on the rocks and sand everyday (which generally wouldn’t significantly regenerate until the following day vs an hour like before.

Was tired of having the eyesore of my UV running out of the display so I got a big bottle of an isochrysis culture and started dosing daily. The Dino’s completely disappeared after a week. Hesitant to attribute causation, but it did seem to line up to suggest a relationship. I didn’t verify but the dinos seemed to be Ostreopsis or possibly Coolia. 
 

If the Phyto was only helping by boosting pod populations, I wouldn’t have expected to see such a big change in only a week.

Thanks for sharing your experience.   My dino's are still with me, but they don't own me!    

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On 2/4/2023 at 1:35 PM, obrien.david.j said:

We're out of town this weekend.  Will update after getting home tomorrow and breaking out the microscope.

Got home, broke out the microscope.   Three things to report.

1. Nanno culture looks clean to me.  No movement, everything visible was uniformly round.

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2.  I still have Dinos in the gravel of the display tank.   I'll have to go look up how to ID them again, but they're there.   Also see nice amount of diatoms (rice shapes), maybe I should restart silica dosing again. 

My last ICP test (Jan 2nd) said I had 270 µg/l  of silica in the water, so it's not depleating back to zero.

 

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3. Found a really cool creature in the gravel.   Got a nice video of it moving around.  Looked like it was munching on something when I found it. 

image.jpeg

 

 

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10 hours ago, Lexinverts said:

Have you tried dosing Iron? I was having an issue a while back and it turned out that I was deficient in Iron. Within 3 days of dosing Iron, the dinos disappeared. My latest challenge is dealing with Cyano.

According to Mack's "dealing with Dinos", high Iron feeds Dino.  But, as far as I can tell - defeating dinos is actually about keeping a healthy population of other organisms, that outcompete them.   Dosing iron for you, could have revived some other population.

In my case, I dose iron daily.  It's part of the reef moonshiners recommendation of daily elements.  Their calculator suggests 2 drops daily, I've done tests a 2/day and 4/day and even a pop up to 6/day - no dino difference.

Dealing with Dinos:

Quote

10. Avoid supplements and salt mixes that are known to add iron to the system.  These can also fuel dinos as well.

...

If water changes are required we would recommend using a high quality salt mix that does not contain high levels of iron, one of our team recently had an ICP completed based on a clean salt mix using Tropic Marin pro, the snapshot below shows the results elevated levels of iron, manganese, Lithium Barium, it is believed it’s the high iron content within fresh salt water mixes causes the dino population to increase

...

We would recommend currently doing water changes using Red Sea’s blue bucket salt as the general consensus is this doesn’t have high iron levels. The below link is to a post on Reef2Reef where Jason2459 has put together a number of tests based on fresh salt mixes and posted the relevant ICP results of these: 

 

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Harvest #29 complete (Feb12th)

  • Continuing to add mid-grow fertilizer kicker.   (2nd dose, after about 1wk) Harvest is back to dark-dark grow of the past

New Things this time

  • I've noticed a difference in culture color (density) for a while
  • Bought a Secchi Stick to put some numbers between cultures
    • Lightest is Jar 1 - density soore of 18-19
      • I posted earlier went kind of clear after Kicker Fertilizer
      • Shares an air supply with another jar.   (Jars 1&2 share, Jars 3&4 share)
    • Darkest is Jar 5 - density score of <10
      • Newest jar, bought used and is fed by it's own dedicated air pump
      • I've noticed each time I harvest, this jar has 1/2-1" of evaporation.  Others dont.
      • The color of led light has always looked a little different to me too.
  • I've been feeding the Coppod culture differently this cycle.  Been dirtying the water every ~4days, using PhytoFeast.   They're growing, so just harvested their new Phyto into 16oz containers, instead of swapping the pod culture water with ~2 liters of fresh phyto.

Trying an experiment,  Does airsupply matter?

  • Swapped Cap/Air Supply between Jar 1 & 5   (Jar 5 had black airhose, now Jar 1 has the black airhose)
  • Made sure to use starter culture from the same mother Jar (used 5, because it was darkest)
  • No other differences between the setups.   
  • We'll see if density and evaporation moves from Jar 5 --> Jar 1.  Stay tuned...

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Thanks to @Eatfrenchfries, I've added a new phyto strain and new pod strain.   

Phyto:

Dumped one of the existing culture jars, and restarted with Tetraselmis.   Moved about 24" away from other culture jars, as a simpleton attempt to reduce cross contamination risk with other cultures.    Going to grow both cultures the same way.  Same timeline, lightcycle, Fert cycle, etc.

Copepods:

  • Tisbe (Original Culture, from Poseidon Reef)
  • Tigriopus (new culture from @Eatfrenchfries)

Started a new gallon jar for the Tigri pods.   So capacity increase from six "gallon" jars, to seven.   

Additional update on status of Tisbe pods.  I've moved to feeding them every ~4days with PhytoFeast, and it's working.   I'm convinced I've starved my culture in the past, leading to my poor results.

Side topic, also got a sample of dragons breath and learned a new technique of tieing it near the surface with fishing line.   

 

 

 

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On 2/13/2023 at 8:35 AM, obrien.david.j said:

Trying an experiment,  Does airsupply matter?

  • Swapped Cap/Air Supply between Jar 1 & 5   (Jar 5 had black airhose, now Jar 1 has the black airhose)
  • Made sure to use starter culture from the same mother Jar (used 5, because it was darkest)
  • No other differences between the setups.   
  • We'll see if density and evaporation moves from Jar 5 --> Jar 1.  Stay tuned...

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Follow-up on experiment.  Two days ago I gave another mid-cycle fertilizer kicker.   Guess what, it looks like the air supply matters.   The Black hose lid (originally on Jar 5, but moved to Jar 1 for the experiment) has a dedicated airpump.  The others are all tee'd off one output.

Looks like Jar 5, with the tee'd air supply cap has turned lighter.  While Jar 1, with the dedicated/stronger air supply, has embraced the fertilizer and stayed dark. Guess what I'll be figuring out how to change... (air supply upgrades for the other jars)

@Eatfrenchfries - did you say you use dedicated USB air pumps, per culture container.  Which ones, where did you get them?

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Currently using portable USB powered air pumps from DHgate at $1.78 a unit and I've had them for over a year with no issues. Paired them with a USB charging splitter to manually control each one.

I bought 30 my first time around and I'm using almost all of them continuously for projects. 

They are also available on Amazon for a few dollars more. 

Screenshot_20230220_131801_Chrome.jpg

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