Culture Expansion

[Eatfrenchfries]

Life just happens. Recently finished my contractor position in Portland so I have a lot more free time on my hands. If anyone is hiring let me know haha.

Decided to dedicate a bit more time to get culture production up because meeting demand + working full time + school is just a nightmare. 

Realized that I was tapping out some of my own cultures to keep up with local demand after my own obligations so I made a dedicated system for zooplankton to get around that.

Current production is about 65 gallons of zooplankton + 45 gallons of phyto split across three systems. 

Master Collection - dedicated glass flasks + air filters + air supply + controllable LED diodes + handmade F/2 for the continuous masters. Then there's a 500mL vial with filters for each cultures I swish daily to keep alive in case of scorched earth contingency. Running 3.5 years (1.5 with no critical incidents). Harvest every 5-7 days. Room temp is maintained. Minimal TLC

P. Feed Supply (45g)- phytoplankton + zooplankton for feeding. Also used to grow things on a whim as needed since everything here is interchangeable: culture vessels + T5 + air filters + dedicated air pumps + 1/2 batch 1/2 continuous. This is the system I kept tapping which took a while for some of the less prolific pods to get going again. Harvest every 7-10 days.  Temperature runs amok but nonissue because of diverse feeds. Running 7 months. I keep duplicates of all supplies/vessels to be able to do it on the fly and clean later. 

Surplus Supply-  Last but not least is the newest addition which I will be sharing part of. One of my mentors in the area graciously thought of me when they had a spare 30g RO storage. Thank you very much my good sir. With this I'm basically using a portion of my Primary to seed this vessel and keep one dominant species from taking hold each week. The original plan was to have the vessel producing within a month but after successfully harvesting my pod cysts for the first time I've been able to cut that time in half. This will be the only vessel heated in my whole production line. The reason why is because pods will reproduce faster, but if they're reproducing faster than I need to keep up the feed or they will crash. Will not be changing with fresh saltwater often because I am collecting the detritus to feed my purple non sulphuric bacteria for snow. Hence why this system has 16 gallons of phyto dedicated to keeping #s but not enough to bloom. Leftover phyto from all my other systems will be pooled to do greenwater changes. 

Essentially just a 30gallon container I added a low air supply but still circulates though sometimes theres *no air at all hehe*, plenty of phyto + pods + heater. 

Ambient light comes from the phyto system which are 1/2 gallon glass milk jugs with two snug air holes for rigid tubing(air pushes in so anything going in is pushed out). LED grow light in the day spectrum. Harvest every 5-7 days for phyto and still haven't harvested the zooplankton yet but soon.......

Would love to post videos showing the zoo cultures but sadly unable to. Pictures are a bit more difficult to capture due to their size. 

Just wanted to show that big or small it's all the same. Just the amount of sterilization involved changes. Obviously I go the extra mile to keep things pure since some strains to replace cost upwards of a $100 not to mention its more than a splash to start.

Phyto shown in the pictures are tetra, iso, and dunaliella. Pods in system are apocyclops, tigriopus, tisbe, parvocalanus (just for show, they will either dominate if I maintain feedings of Thal/Iso or they'll die out from not enough). 

The other picture with the coke bottles was mixing enough F/2 to last me a couple months. I usually just make bulk without silicates and then I add in water glass mixed with RODI to the culture water. 

Also just something I thought was cool that a few people asked about when they came by were these reef "mugs" my girlfriend wanted for her nano tank. I designed a reef viewer with a hook handle that can hook on the side of a tank. Made a few that sold quick so if there's a demand I'll make some more. Currently in 3inch and 6inch sizes. 

 

 

 

Edited March 9 by Eatfrenchfries

[obrien.david.j]

I've been thinking about your home made F/2 fert.   Have you ever watched Cody's Labs.   He made F/2, twice, and an outdoor phyto grow system.   Fun videos

 

 

 

 

[Eatfrenchfries]

I am a little familiar with Cody's labs. Running the same premise of algae + pod cultures outside but since I can't track outside contaminants I mainly use them for personal use. Over winter they'll freeze over but come spring the production speeds up on its own. Sunlight only + culture + food source (dead bugs, banana peel, greenwater) Mainly deal with infusoria, synechococcus, nanno, tig pods for outside cultures due to them being able to withstand fluctuations and temperature extremes. Plus they'll usually remain dominant. Tigs will go dormant until temperatures resume. 

(First time running this in Oregon and it's worked when months are warm. Current pods are hibernating. Used to keep this in California year round, just needed to shade it partially)

 

Mainly I make my own F/2 media to have absolute production control. Not something for the faint if heart because one misstep could ruin future batches. Most prefer to buy it dry in bulk and mix it themselves.

Hand making F/2 should only be done if you have an actual need to alter the formula. Doesn't add up to do it yourself when someone else can do it for cheaper and guaranteed. There's too much sterilization involved to keep it pure. Had a buddy of mine start mixing his own and that batch was contaminated during mixing so it crashed all his cultures in short order with some sort of cyano. 

One batch of my F/2 is mixed stronger than most for most phyto application.  Second batch is made with room for silicates. Third batch contains heavy metals + ammonia. Fourth batch contains extra carbon sourcing. Everything is based on the standard F formula. I play with F/2 and F/3 for culturing purposes. 

[John Vinson]

Do you sell any of your F/2 mix as well? 

[Eatfrenchfries]

@John Vinson I would consider in the future but as of right now I am still developing the refinement process not to mention the formula. 

I could possibly give you a sample to try out for a few batches. 

*I do have an autoclave but there's a lot of technical effort involved in this process. I trust myself but even I'm not perfect. 

[Brittany]

Great talking to you today.   Thanks for the information.

[Eatfrenchfries]

I figured I had the microscope dialed in but getting a digital camera just means more tinkering to be done. Promise I'll get better. I used to use a camera mount for my phone but must say a digital camera does help a lot. 

Just some pictures of the pods I'm working with.

I can get the microscope to focus but the distance of the camera to the lens distorts the image. (Gotta love YouTube videos for troubleshooting)

Parvocalanus crassirostris 

Tigriopus californicus 

Tisbe biminiensis 

Apocyclops panamensis 

*want to point out in the image with the parvocalanus copepod that the colored dots stuck to it is isochrysis / Thalassiosira (phyto). Overfeeding phyto too fast can ruin pod molts since it's too dense for the culture water. 

Parvocalanus has been my favorite since it behaves differently than the other pods as a visible adult. They float in the water column and use their antenna to detect motile phytoplankton. Using their short legs they quickly pounce (teleport) on to its prey. 

 

 

[obrien.david.j]

I'll try a few pictures next time I harvest mine.   So far, I'm happy enough with the cell phone mount I have to take pictures.  But we'll see.

[Eatfrenchfries]

I've had a few questions on why my blend varies in color and isn't always your typical rich green blends but closer to brown most of the time. I culture X amount for contracts and Y for myself that sometimes there isn't always an even ratio in the blend. 

Why does it vary in color? I culture different strains of phytoplankton so,  red + green + brown = brown 

Microalgae Strains: 

Isochrysis - a golden brown 3-8 micron size motile microalgae that is high in fatty acids making it an excellent feed source for filter feeders and gut loading zooplankton. 

Tetraselmis - bright green that get up to 10 -14 microns motile microalgae that is rich in amino acids perfect for feeding. 

Thalassiosira - brown beige ranging in 4 - 32 microns. This is a diatom that requires the addition of silicates to thrive. Beneficial in cycling tanks faster and highly variable as feed for marine organisms due to its large size. 

Nannochloropsis - emerald green 2 - 5 microns nonmotile microalgae. Nutritious feed but has a tough membrane for pods. Excellent source of pigments. 

Porphyridium - ruby red 6 - 10 microns nonmotile microalgae. Highly nutritious (growth) and contains phycoerythrin pigment used to make red filter feeders color pop.

Rhodomonas - strawberry lemonade 9 - 36 microns nonmotile algae. Highly nutritious (growth) feed. Good for photosynthetic organisms and gut loading. 

Symbiodinium (zooxanthellae)- brownish dinoflagellates 6 - 13 microns harbored by photosynthetic organisms like corals, clams, and anemones. Potential to restore coral bleaching. Used as a technical feed for cryptic organisms that require photosynthetic exudates. Currently working with two lab grade and two I harvested myself. 

Synechococcus - forest green cyanobacteria 0.1 - 1.5 microns. Bulk abundant feed source that is fast producing. Used to cover initial larval stages across the board and a staple feed. Somewhat motile, it is believed to possibly compete with red cyanobnacteria found in aquaria. 

 

Diversity in cell size helps cover varying stages of zooplankton life while nutritional profile maximizes yields. I'll say covering cell size is more important than providing optimum nutrition when going for reef bugs. They're pretty hardy when it comes to food if the size is right. Nutrition helps when gut loading for feeding and keeping densities up without competition. 

(Pictures included are 7 of the microalgae strains listed above and one of the bulk pod culture after being freshly fed. )

Bulk Pod Grow after being fed

Symbiodinium #2

Thalassiosira

Nannochloropsis

T

Tetraselmis ( I know the post its say nanno but they've been repurposed, different green)

Porphyridium

Isochrysis

Synechococcus (grown partially outdoor with manual stirring)

Edited 20 hours ago by Eatfrenchfries

[obrien.david.j]

My Nanno and Tetra continue strong growth.  I hit the 1yr mark of growing Phyto in March.