Culture Expansion

[Eatfrenchfries]

The 2 liter soda bottle is just how I keep the culture. The only circulation is when I harvest 1/5 phyto and replace with RODI. This display bottle is place under T5s. My bulk culture is suspended via shake plate.

 

The environment this algae comes from regularly dries up so making it rain is the easiest way to suspend the algae

[Eatfrenchfries]

Just wanted to wish everyone a happy new year. 2023 was an eventful year that taught me a lot.  

Thank you to everyone in the community.

Special thanks to everyone who has supported my development. Would not be here if not for all of you. 

 

 

A lot of people ask me about my "setup." Tis not much but this is my primary workstation. 

Pretty much decided to see how much I can fit into my apartment without my girlfriend killing me. 

(Picture: zooplantkon production on the left rack / computer and more zooplankton behind it / 90gallon macro growout t5 top, led actinic bottom)

Zooplankton production except rotifers and brine. Buckets have hardier species. Clear vessels contain pods that require fresh food daily. Fresh and salt zooplankton are here.

7 marine pods,1 amphipod, 2 rotifers, 2 fresh pods, 2 water fleas, black / white worms, brine (baby + adult) in full production.

Currently developing the capability to culture 2 new marine pods.

Top of the rack. Crayfish, black worms in the tubs. Buckets contain rotating pod cultures that I experiment with. Vases under the poster contain freshwater zooplankton (moina, daphnia, cyclops, diaptomus). Out of view. Brute trashcan for amphipod rearing. 

Microalgae culture station. Masters on top. Bulk on bottom are rotated weekly to keep water down. Currently maintain 15 distinct cultures. 

Currently only two breeding projects. 

Halocaridina rubra (top) and Pseudomugil cyanodorsalis (bottom). 

Got a little saltwater hydroponic set up.  Mainly red / black / orange mangroves, but there is a bit of seapurslane grown in a coc coir. Stocked with guppies / ghost shrimp / purple and green bacterial mats @    1.024 salinity

 

 

Bacteria, rotifers, and brine are kept in a separate "office" room. Helps keep contamination down. 

 

 

Slowly learning to not be pulled in different directions. Just do what makes me happy. 

[Krux]

your lady is a saint.  Amazing what you have been able to assemble there!

[Eatfrenchfries]

I

Running out of refrigerator space for all my cultures. Started spinning some at 1500 RPM for 10minutes to condense for feeding. 

The use of sodium alginate at 0.38 grams per liter has proven to be highly effective in keeping suspension. Realized that sodium alginate won't work on live phytoplankton unless it is quite dense. 

 

Currently running these at 1L masters and 3 - 5 gallon bulk. I can only store 500mL of each culture as a backup and 7G of blended feed. But now that I can condense phyto I can store 3 - 5 times the amount without breaking a sweat. 

-Nannochloropsis 

-Tetraselmis 

-Chlorella

-Thalassiosira 

-Isochrysis 

-Pyrocystis 

-Phaeodactylum 

-Chaetoceros 

-Porphyridium 

-Rhodomonas

-Symbiodinium A

-Symbiodinium B

-Dunaliella 

-Spirulna

-Scenedesmus 

-Haematococcus

-Pavlova

-Synechococcus 

Couldn't have this many cultures running without @LavenderTi and the carboys. This was the tunring point that got me deeper into the rabbit hole.

Biggest downside is that the phyto cells lose motility quite rapidly this way. But the consistency is akin to phytofeast.

***PNSB when suspended in the alginate will consume it over time. Not recommended. But it seems to have no problem losing suspension and being motile if concentrated.

 

 

Shoutout to @CuttleFishandCoral for the awesome sea apple. This made my week. 

 

 

 

Thank you @Zach0225 for the CO2 tanks. My PNSB production takes a lot of my CO2 (20lb tank). But now I have enough tanks (2x5lb) to resuscitate phytoplankton cultures. 

Photo pictured above is CO2 addition bringing back a Chlorella culture. 

My hope is to fine tune a few of my finicky cultures to be bullet proof. So as not to renew them yearly. 

The CA reactors will be used to make a effluent reactor that I can use to grow more PNSB. 

 

[Krux]

Any use for stainless steel storage containers?  Once I saw the CO2 canisters it reminded me that I have several 5 gallon kegs from home brewing in my garage.  They can be sterilized and used for liquid storage of some sort...

Let me know if you have any interest, and look up Cornelius keg for more info.  I have between 6 and 8 you are welcome to play with.

[Eatfrenchfries]

Scrapped the idea of using glycerin or sodium alginate to keep the concentrates in suspension. 

Took a step back and realized centrifuge wasn't enough to layer phyto cells on one another and maximize storage space. 

 

1. Run phytoplankton through centrifuge 

-focus on removing water content

- keeping cells intact

- maximize storage 

2. Use 1um mesh to drain water further and create paste 

(5um did not work as some species are less than 2um)

 

*two methods from here

3A. Paste is stored in 20ml glass droppers and refrigerated for up to 7 days of use

- cells are intact therefore maximum nutrients

- motility is maintained for 4 - 7 days (some species do better than others)

- easy to store and use

- can be cultured

3B. Paste is flash frozen by making a dry ice sandwich with wax paper. (Thinner the better)

- cells are mostly intact (70%)

- some nutrition lost

- backup feed that can be stored 1 - 2 years for feeding purposes

- unable to culture

- easy to rehydrate 

 

 

 

 

Polyp response with frozen paste

 

[obrien.david.j]

Thanks for laying out your thought process and progress.  

As you know, I'm culturing Nannochloropsis Oculata and Tetraselmis.   I'm harvesting straight into 32oz container(s), which takes me 2-3wks to feed them all to my tank.   Stored in refrigerator that long.   Am I still feeding with max(or close) nutrients, and good motility?

 

[Eatfrenchfries]

1 hour ago, obrien.david.j said:

Thanks for laying out your thought process and progress.  

As you know, I'm culturing Nannochloropsis Oculata and Tetraselmis.   I'm harvesting straight into 32oz container(s), which takes me 2-3wks to feed them all to my tank.   Stored in refrigerator that long.   Am I still feeding with max(or close) nutrients, and good motility?

 

If the phytoplankton is refrigerated and agitated (daily is ideal) then it will keep nutrition. Up to 4 months if it is a single strain. The cold will slow the metabolism. 

 

Motility can diminish if not agitated or if multiple species are present in the storage vessel. 

Nannochloropsis is nonmotile. The higher the density, the faster it will settle. 

Tetraselmis is motile, this means that once it loses suspension it will no  longer be motile and cannot be resuspended easily. 

 

*motility only matters in rearing certain organisms. Pelagic pods need motile plankton as a large number of them are ambush predators. 

-if algae is allowed to settle at the bottom of the vessel then all the cells will begin to "suffocate

 

 

Some species of phytoplankton are easier to store than others. 

Isochrysis has a terrible shelf life in the refrigerator. Better to be room temp. 

Rhodomonas likes the cold but will degrade immediately and begin to stress making it difficult to store backups. 

[obrien.david.j]

1 hour ago, Eatfrenchfries said:

If the phytoplankton is refrigerated and agitated (daily is ideal) then it will keep nutrition. Up to 4 months if it is a single strain. The cold will slow the metabolism. 

Motility can diminish if not agitated or if multiple species are present in the storage vessel. 

Thanks.   In my case, I keep the species separate.    What I'm not doing today is agitating regularly.  I'll fix that.