phytoplankton culture settling

[fryman]

I have the 1 part Guillard’s F2 and it has begun to show some dark pieces of debris in it. Is this some kind of normal precipitation or should I pitch it. I don’t think I contaminated it in any way.

Dark debris isn't normal, in my experience. I dunno what it could be. I've heard that 1-part doesn't store as well as 2-part but I've used both with similar results.

No matter how strong i bubble culture, i have habbit that everyday i swirl container a bit, it helps a lot with settling.

I've heard alot of people have settling issues but I'm afraid I don't have much advice to offer. I have little to no settling in my cultures. I used to get crashes but follow a pretty strict protocol now so it's rare for me nowadays.

Does it matter how GREEN a pod culture is. Is just a tint of green enough or should it look more like the Tetraselmus in the attached photo?

The nanno looks good to me, the tetraselmis looks like it was just split. Generally speaking darker means more dense. But alot of things are green so just looking dark green doesn't mean high quality ime.

Sounds like since I have a multistrain culture eventually one will become dominant. Do you culture multiple strains? Is the work worth the diversity between strains?

Do you only dose f/2 right after the split or is it something you do on a more regular schedule?

My usual mix is tisochrysis, tetraselmis, and nannochloropsis. I have tried others but that combo is great for most uses (I grow copepods, rotifers, and artemia). For bivalves (I'm trying flame scallops) I'd add diatoms like Thalassiosira.

It makes a difference but you can get by with a single type sometimes. I've grown rotifers on just nanno and they did quite well. Tigriopus copepods will grow on about anything, as will artemia. Other copepods didn't grow for me on a single feed type. But you could feed one type live and supplement with a preserved mix. There are other ways to go.

It was difficult for me to get multiple types started at first but once I had them it's not significantly more work having 3-4 cultures going vs. Just a single one.

I only add f/2 when splitting.

 

[TWYOUNG]

Dark debris isn't normal, in my experience. I dunno what it could be. I've heard that 1-part doesn't store as well as 2-part but I've used both with similar results.


I've heard alot of people have settling issues but I'm afraid I don't have much advice to offer. I have little to no settling in my cultures. I used to get crashes but follow a pretty strict protocol now so it's rare for me nowadays.


The nanno looks good to me, the tetraselmis looks like it was just split. Generally speaking darker means more dense. But alot of things are green so just looking dark green doesn't mean high quality ime.


My usual mix is tisochrysis, tetraselmis, and nannochloropsis. I have tried others but that combo is great for most uses (I grow copepods, rotifers, and artemia). For bivalves (I'm trying flame scallops) I'd add diatoms like Thalassiosira.

It makes a difference but you can get by with a single type sometimes. I've grown rotifers on just nanno and they did quite well. Tigriopus copepods will grow on about anything, as will artemia. Other copepods didn't grow for me on a single feed type. But you could feed one type live and supplement with a preserved mix. There are other ways to go.

It was difficult for me to get multiple types started at first but once I had them it's not significantly more work having 3-4 cultures going vs. Just a single one.

I only add f/2 when splitting.

My tetraselmus starter cultures from Algae Research Supply were much lighter than nanno. Those were split three days ago. Each time I split them they go clear for a day before they recover and start growing. They never get near as dark as my nanno. but I confirm they're alive via scope. My main concern is the color of my pods which were split today. How green do they need to be? Is the slight green tint they are currently dark enough, or should I add more phyto.?

 

[pmcmahon]

I've heard alot of people have settling issues but I'm afraid I don't have much advice to offer. I have little to no settling in my cultures. I used to get crashes but follow a pretty strict protocol now so it's rare for me nowadays.

Would you be willing to share your routine?

 

[reef_1]

I do normal fertiliser dosing, cant be bothered about f/2.

You can measure nitrate levels in the culture water, to see if phyto has food.

In phyto water I would use an airstone.

Imo when splitting make it much lighter until it matures in vaguely in a week, then you have maintenance only weekly once.

Tetraselmis culture should be dark green.

I would make pod cultures with a bit more tinted water, so that you can better following if they have enough food/phyto based on color.

Cultures not growing are infected, crosscontaminated cultures, or bad water, nutrients level (too much media can kill a culture as well).

Phyto cultures are quite resilient and can be restarted from month old fully settled stinky sludge, so you can be more brave when splitting to minimize maintenance.

 

[fryman]

My tetraselmus starter cultures from Algae Research Supply were much lighter than nanno. Those were split three days ago. Each time I split them they go clear for a day before they recover and start growing. They never get near as dark as my nanno. but I confirm they're alive via scope. My main concern is the color of my pods which were split today. How green do they need to be? Is the slight green tint they are currently dark enough, or should I add more phyto.?

Tetraselmis should get darker than that, mine gets about as dense as my nanno.

I personally would add more phyto to those pod cultures.

 

[fryman]

Would you be willing to share your routine?

For 2-liter cultures I clean or replace the bottle and airline every 7-day split. First with vinegar or CLR, followed by H2O2 or 1:10 bleach. Media (fertilized saltwater) is sanitized by UV and/or bleach. Air is filtered by air pump in vacuum bag in a sealed enclosure.

 

[reef_1]

For 2-liter cultures I clean or replace the bottle and airline every 7-day split. First with vinegar or CLR, followed by H2O2 or 1:10 bleach. Media (fertilized saltwater) is sanitized by UV and/or bleach. Air is filtered by air pump in vacuum bag in a sealed enclosure.

That would be too much work for me I do it in pet bottles (soda bottle), whenever I feel I just throw them away and have a new ones.

 

[TWYOUNG]

Tetraselmis should get darker than that, mine gets about as dense as my nanno.

I personally would add more phyto to those pod cultures.

Added more phyto to pod cxs. I'm curious why my tetra is so pale. The starter culture I received was also much lighter than my nanno. Each time I split it, (including the initial time), it loses all color for about 24 hours until it begins to grow darker again. I've monitored it all along with a scope and it's definitely alive and moving. Any suggestions on how to get it to grow darker?

 

[Smoke-Town]

Phyto cultures are quite resilient and can be restarted from month old fully settled stinky sludge, so you can be more brave when splitting to minimize maintenance.

Is this statement true?

For 2-liter cultures I clean or replace the bottle and airline every 7-day split. First with vinegar or CLR, followed by H2O2 or 1:10 bleach. Media (fertilized saltwater) is sanitized by UV and/or bleach. Air is filtered by air pump in vacuum bag in a sealed enclosure.

If the first statement is true, why would you need to do all this?

I'm only asking because I have been trying phytoplankton cultures recently and am having issues. The pods are easy but the phyto settles and smells like spoiled veggies. I have nice glass I'd prefer to keep them in... I like the laboratory look... really don't want to use plastic garbage bottles and replace stuff every week.

 

[TWYOUNG]

Is this statement true?

If the first statement is true, why would you need to do all this?

I'm only asking because I have been trying phytoplankton cultures recently and am having issues. The pods are easy but the phyto settles and smells like spoiled veggies. I have nice glass I'd prefer to keep them in... I like the laboratory look... really don't want to use plastic garbage bottles and replace stuff every week.

I've been quite successful for about 18mo now. I don't filter the air going in but I soak the 1 gal glass jars I use overnight in bleach solution, air dry then rinse out with rubbing alcohol. I haven't always done it but several months ago I bought an automatic tea/coffee pot with a glass pitcher so I can boil my fresh sw before using. I have a large mixing station and sw is sometimes stored for weeks.

 

[fryman]

Is this statement true?

If the first statement is true, why would you need to do all this?

I'm only asking because I have been trying phytoplankton cultures recently and am having issues. The pods are easy but the phyto settles and smells like spoiled veggies. I have nice glass I'd prefer to keep them in... I like the laboratory look... really don't want to use plastic garbage bottles and replace stuff every week.

You will find a whole range of opinions here on how important sanitizing is, and/or how much is necessary. My protocol is based on standard practice in the aquaculture industry. But at the hobbiest level you will not often see people use best practice. Which is understandable.

Actually, I agree with the post above about keeping a fridge backup. It's most useful imo as a backup, but in my experience can also help "cleanup" a dirty culture. I'm not certain why but think maybe alot of bacteria contamination dies in the fridge, allowing the phyto to "take back" a culture when you restart.

However, it is not 100% effective and some contaminating organisms will survive a period @ 3-5C just fine. Also, it takes awhile for a fridge backup to recover, and so it's not ideal if you have to revert to backups all the time.

And finally, not all phytoplankton types survive storage at 3-5C. In my experience, isochrysis/tisochrysis and rhodomonas cannot be stored in the fridge more than a few days. So if you want to grow those types, your sanitization protocol is more important.

Glass labware is great for growing phyto, just keep it clean. I use 4L Erlenmeyer flasks for my non-fridgeable backup cultures (e.g. isochrysis).

 

[fryman]

I've been quite successful for about 18mo now. I don't filter the air going in but I soak the 1 gal glass jars I use overnight in bleach solution, air dry then rinse out with rubbing alcohol. I haven't always done it but several months ago I bought an automatic tea/coffee pot with a glass pitcher so I can boil my fresh sw before using. I have a large mixing station and sw is sometimes stored for weeks.

FIltering the air is most important if you keep multiple types of phyto to prevent cross-contamination. Before filtering the air, I found that eventually everything turned into nanno or synechococcus.

 

[TWYOUNG]

You will find a whole range of opinions here on how important sanitizing is, and/or how much is necessary. My protocol is based on standard practice in the aquaculture industry. But at the hobbiest level you will not often see people use best practice. Which is understandable.

Actually, I agree with the post above about keeping a fridge backup. It's most useful imo as a backup, but in my experience can also help "cleanup" a dirty culture. I'm not certain why but think maybe alot of bacteria contamination dies in the fridge, allowing the phyto to "take back" a culture when you restart.

However, it is not 100% effective and some contaminating organisms will survive a period @ 3-5C just fine. Also, it takes awhile for a fridge backup to recover, and so it's not ideal if you have to revert to backups all the time.

And finally, not all phytoplankton types survive storage at 3-5C. In my experience, isochrysis/tisochrysis and rhodomonas cannot be stored in the fridge more than a few days. So if you want to grow those types, your sanitization protocol is more important.

Glass labware is great for growing phyto, just keep it clean. I use 4L Erlenmeyer flasks for my non-fridgeable backup cultures (e.g. isochrysis).

I love to see a pic of your Erlenmeyer flask setup. My family thinks mine looks like a lab but I imagine you've got me beat big time!

 

[TWYOUNG]

FIltering the air is most important if you keep multiple types of phyto to prevent cross-contamination. Before filtering the air, I found that eventually everything turned into nanno or synechococcus.

I only grow tetraselmus but I'm curious how that contamination occurs. Do spores of some sort travel back up the airline tubing?

 

[Stephers]

I only grow tetraselmus but I'm curious how that contamination occurs. Do spores of some sort travel back up the airline tubing?

It doesn't take a lot of cells for them to contaminate and then start growing and outcompeting. The bubbling on the cultures is vigorous and I'm sure some cells find their way out if allowed. I've had a lot of luck using airlocks on my containers (used in wine/beer brewing). I keep 4 species (tetra, nanno, rhodomonas, T-iso) directly next to each other and haven't had any contamination issues (knock on wood!!). The airlocks allows gases to leave but nothing to enter and contaminate.