Culturing Phytoplankton Experiment

Dilan Patel

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So I have been wanting to culture my own phytoplankton for awhile now. Having just made my own saltwater mixing station I have a little extra room where I can go about doing this culturing process. Being that about 16oz at a LFS cost me about $40 this could be a nice money saving project that could benefit my reef. I have ordered and or had all the supplies needed. I will be ordering the phytoplankton strain nannochloropsis as well as the f/2 fertilizer from amazon. I will be using a clamp light and a tetra air pump to run this. The lights will run for 16 hrs and off for 8. Unfortunately I will be unable to order the phytoplankton until I get back in town next week I hope to get this project rolling. Right now I am running a temp test for the lighting source I am using to hopefully have a temp around 78 degrees. I am planning on either using a 1 gallon pickle Jar I have as my only culture or do two cultures of .5 gallons each to give me a back up culture still deciding. The gallon would give me about .5 gallons to use on my tank and the other half would be used to start the next culture. I will be dosing 5ml per 20 gallons so about 50ml for me to start which equates to about 700ml for 2 week time period which by then my next batch should be done ( should be done within 7-10 days but 14 days is worst case scenario). Being that a half gallon is about 1900ml I will in theory have extra I wouldn't mind having a few people test it out and see if they like the quality and give feedback. I may or may not get the microalgae density stick so I can get a rough estimate of the cell density. I hope you guys enjoy seeing this project documented as well as give any advice!

Starting Equipment:
Clamp light 8in
60 W LED Soft white (will probably change this to a 5k spectrum like daylight)
Pickle jar(1 gallon) or 2 1/2 gallon containers
Tetra 60 gallon air pump(dual outlets with air control and backflow prevention)
Rigid airline tubing attached to regular airline tubing(no airstone)
Alcohol to disinfect everything. Very important to keep this culture sterile and workspace as any contaminants has a possibility of crashing the culture
Saltwater would be 1.019 Sg
Couple Pics:
IMG_1031.jpeg


Temp Testing before starting Culture
IMG_1032.jpeg
 
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Peace River

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Although I don't have any phyto cultures going right now, from past experience I would suggest that you consider keeping 2 or 3 cultures running because crashes will happen. Good luck on the project!
 

Eagle_Steve

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Although I don't have any phyto cultures going right now, from past experience I would suggest that you consider keeping 2 or 3 cultures running because crashes will happen. Good luck on the project!
This right here. I keep 4 cultures going. A crash will happen, it is just when.
 
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Dilan Patel

Dilan Patel

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So instead of the pickle jar I will run two cultures at about 1/2 gallon each or if I decide to go with another gallon culture. I think two should give me a backup if needed. I really don't need more than a gallon so I was thinking maybe just go with the standard 2 x 2liter bottles. As for lighting what spectrum do you guys recommend?
 
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Eagle_Steve

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So instead of the pickle jar I will run two cultures at about 1/2 gallon each or if I decide to go with another gallon culture. I think two should give me a backup if needed. I really don't need more than a gallon so I was thinking maybe just go with the standard 2 x 2liter bottles. As for lighting what spectrum do you guys recommend?
I use cheap led strip lights from amazon. Wrap them around my bottles and let it go. I just the 6500k ones and all seems to grow well.
 
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Dilan Patel

Dilan Patel

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Phytoplankton came a day early so I just finished starting the first batch of phyto. Basically I am now using 2x2-L bottles rinsed with Vinegar and then hot water to help steralize the bottles, my hands as well as any equipment used. I was unprepared for them arriving early so I will sit on the second bottle I got in case something happens to this batch I can immediately try again. But basically I split one 16oz bottle between the two culturing bottles. I Then topped them off with RODI saltwater mixed to 1.018-1.019 which is in the range they recommended. I then added 1ml to both bottles of F/2 and now they are bubbling to keep the phyto suspended with 2 60W 5k bulbs on for 16hrs. The mixtures are still pretty dark green but def lighter than before. My goal is to get to the dark green I started the cultures with. Here are a couple pics (not the prettiest but it will work for now). Now for splitting the culture do i need to wash the current bottle then reintroduce the culture or can I just pour out 3/4 of the product and then backfil it and add fertelizer?
IMG_1089.jpeg

IMG_1091.jpeg
IMG_1092.jpeg
 
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Dilan Patel

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had a little hiccup I noticed on the 10th. The color was not darkening as much as I expected come to find out I only fed .1ml instead of 1ml lol. Well luckily I did not starve the culture yet so I added 1ml to each bottle and within that day the phyto was darkening. As of rn the pohyto is getting super dark and it may even be ready to harvest in the next few days. I then plan to split the culture and leave about .25-.5 of 2L left in each bottle to restart the culture and also add 2 more bottles of phyto to start culturing.
 

MohrReefs

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When its emerald green, harvest. I grow my own too and I noticed that once it hits this color the culture is done and if I wait another day it's usually followed by a crash. I suspect it's because they've consumed all of the fert. I also noticed the culture does better in a 1.025 salinity. You can also use this to grow copepods too. I do both.
 

MohrReefs

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Also, make sure to clean everything with alcohol in between harvests. Make sure the alcohol dries completely. Dirty equip crashes cultures as well as residual alcohol.
 
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Dilan Patel

Dilan Patel

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So your saying I need to transfer the split harvest into a new bottle? When I clean/sterilize the stuff I used vinegar and then rinsed it out with Tap water to get rid of the residual vinegar. How does this color look? I am trying to get the color to match my starter culture or is that too far? Thanks
IMG_1124.jpeg
 

MohrReefs

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Me personally, I harvest when the culture is the color of yours. I make a gallon every 3 or 4 days. I'm up to my ears in phyto.
I'd rinse with distilled water just incase (phyto is sensitive to anything in the water), but vinegar can definitely be used for cleaning. Looks great, you seem to be doing fine, soon you'll have more than you can even use.
 
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Dilan Patel

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Thank you! How much fertilizer do you feed? Ive read that 1ml is standard for 2L bottle but I saw people put up to 5ml per 2L. Do you know if there're any risk to overdosing the fertilizer other than wasting product? Im guessing even if I do harvest a bit earlier that is better than bottoming out the nutrients causing a crash. I am going to let it run today and maybe tomorrow putting me at about 4 days with the proper dosage of fertilizer as I did mess up with the fertilizer dosage starting out ran about 4 days with just .1ml in each bottle lol. I may invest the couple bucks into the secci principle stick just to see how dense the phytoplankton is. Thanks for the advice!
 

MohrReefs

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Thank you! How much fertilizer do you feed? Ive read that 1ml is standard for 2L bottle but I saw people put up to 5ml per 2L. Do you know if there're any risk to overdosing the fertilizer other than wasting product? Im guessing even if I do harvest a bit earlier that is better than bottoming out the nutrients causing a crash. I am going to let it run today and maybe tomorrow putting me at about 4 days with the proper dosage of fertilizer as I did mess up with the fertilizer dosage starting out ran about 4 days with just .1ml in each bottle lol. I may invest the couple bucks into the secci principle stick just to see how dense the phytoplankton is. Thanks for the advice!
I bought my system from Poseidon Reef Systems, they include a part A & B fertilizer, I use 2ml of each with 3/4 cup of phyto and the rest of the solution with salt water. If the salinity is a bit high I'll add fresh RODI to drop it down to 1.025. I have my jars wrapped in LED lights and I light it for 16 hours and dark for 8.

To be honest, you could just do a couple test runs with different amounts of fert and just log your results until you find which method works best for you and give you the most yield without a crash. It takes little phyto to start a gallon of culture.
 

MohrReefs

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20200318_192406.jpg


Heres my set up, phyto grows and feeds copes, left over phyto is stored for future use and tank feeding. All stored in fridge when not in use.
 

Aqua Man

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Mercer of Montana phyto is what I’m culturing too. So far I haven’t been able to get as dark as the original. Let it go to 10 days from splitting and was closer.

When it’s time to split, I put one whole culture in containers to store in fridge and use.
Completely sanitize that container and hose. Then dump the other culture into that one. Sanitize the now empty one. Pour back half and top off with salt water. 1.019 SG.

Still very much a novice of phyto. Following your adventures!
 

MohrReefs

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Cool thanks. May have to pick your brain about copepod culturing if I start wanting to do that.
Yeah, shoot me message if you ever starting growing copepods. My mandarin has unlimited food and it saved me 20$ a week in copepods. You get your money back fast.
 
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Dilan Patel

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Just split the cultures. Honestly it looks pretty light. But didn't want to chance starving out the culture. Here are some pics.This time around I put the right dosage of fertilizer lol. I also used the second mercer bottle and I am testing if 1.5ml of fertilizer makes a difference compared to 1ml. 2L Bottles 1&2 are the mercer batch while 3&4 are my split batch. 1&3 have 1.5ml of fertilizer while 2&4n have 1.0ml. Hopefully these come out nice and dark :). Everything was sanitized including the previously used bottles.
IMG_1130.jpeg
IMG_1129.jpeg


New Batch:

IMG_1131.jpeg
 
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Dilan Patel

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Just split my batch after not seeing much color change. TBH I am a bit confused about a couple things. 1 when culturing the phyto with the 1.5ml dose of food appeared darker than the one with 1.0ml. I take them out today and look at them in sunlight and they look pretty much identical. I then compare to my previous batch which looked awfully light while in the closet but they appeared identical as well. Is it that I am comparing them, indirect sunlight as I can place them on my table and can easily tell the difference. I have ordered the secchi stick to see if I am even increasing the cell density(which would correlate to a darker color). Here are some pics today. Considering that they were not growing after the 3-4th which I believe I still did not have enough food. Today I have again increased the amount of food in each vessel with vessel 1 starting out at 2ml of food through vessel 4 having 5ml of food. Im hoping this way I can pinpoint how much food I need to get the darkness I want as well as see a gradual change in color on the next harvest day. I also have noticed in this batch some green particulates floating around. Is this contamination and i need to start over or is this really big phytoplankton lol.

Harvest:05/20/20
IMG_1173.jpeg


Looked like in the closet:
IMG_1172.jpeg


Previous and current batch comparison...can you see a difference?
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IMG_1178.jpeg
 

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