Culturing phytoplankton - Observations

melev

melev

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[(Susan is Flame*Angel from Reef Central)

I wanted to share something I've noticed recently. Susan and I have been trading a few emails as well on this topic.

Early last year I started to culture phtyoplankton, and it went fine for about 6 months. I began to see cyano in my tanks and figured it might be time to restart my cultures. However, it may have been caused by something else (over feeding Golden Pearls for example). I started a new batch but forgot to split it at 7 days. On day 14 I checked on it to split it and it was yellow and decomposing. Argh.

Started a new batch and reread Susan's page and realized I was supposed to split it at 7 days. No problem. However, a few weeks later my culture crashed again. I couldn't figure it out. How did I have success for 6 months and now not?! I ordered more disks from FAF.

Now each week when I harvest, I run the phtyo through 2 coffee filters (doubled-up) to screen out any crap from the bottom of the bottles. It looks like shale, maybe Calcium flakes/deposits? Coffee filters are very slow. I had the screen for harvesting Rotifers in my garage, so I use that now to screen my phyto. Nice and fast and the phyto looks clean and clear.

What I find shocking is this: Last week, I split my cultures at 6 days, and already they were lighter green, almost turning yellow. Susan mentioned that she'd stopped adding Micro Aglae Grow to her medium, and simply poured in 1 ml per 2-liter bottle. I had some culture medium already in 2-liter bottles waiting to be used, so I added 1 ml per bottle in addition to the original solution (1 gal 1.019sg plus 2ml/gal). Within 3 days, my new batch was extremely dark. On day 5 I decided to split it before it crashed. Look at this picture below. The bottle on the left is last week's batch, the bottle on the right is from today. (Which was started from last week's bottle!)

phyto_color.jpg
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What I believe kept crashing my batches was a lack of Micro Algae Grow. I'm going to continue to screen my batches, and use 1 ml per 2-liter bottle to try and maintain this coloration.

The bottle on the left, when measured with the density stick was 2.5 cm, the bottle on the right was 2 cm. Looks like my density is at 19.2million cells or better.]
 
[Ok, I am trying to do this and NEVER seem to get it to get that dark of a green. So, could you write me exact steps? I see what you are saying now is you do the following (let's say you were just starting from scratch).

1. Fill 2-liter with Saltwater.
2. Add 1ml Micro Algae Grow to bottle.
3. Add ? phyto (how much starter culture? where to buy and exactly what)
4. On Day 6, pour half of bottle in new 2-liter (filter through rotifer screen when doing so...assuming plankton is small enough to go through rotifer screen)
5. Add 1ml Micro Algae Grow to new bottle.
6. Fill new bottle with more saltwater to top.
7. Fill old bottle with more saltwater to top. (Do you add more Micro Algae grow to it also?)

Help me! I keep seeing my culture get slightly darker and then reverse. Don't know what the deal is.]
 
[Hi Jody,

You've almost got it right.

To start off from scratch, you fill up the bottle completely, stopping about 1" from the top. It should be saltwater that is 1.019sg - using a refractometer would be ideal.

Using a full disk of Nanochloropsus from FloridaAquaFarms.com, you would scrape the green algae film into your bottle after it has soaked for 24 hours.

Add 1 ml of Micro Algae Grow, also from FAF.

This is a picture of the stuff I ordered (note I only got one disk, but you should just order 2):

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Here's a picture of what I was doing. In the picture, I put 1/2 the disk in each bottle, but I think it would be better to use 2 disks -- one disk per bottle.

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Set the bottles up with the rigid tubing going to the base of the bottle. It should bubble gently, and not be a rolling boil.

Lighting should be on for 16 hours a day, then in full darkness for 8 hours. I have mine culturing in a closet where it isn't annoying. Here's a picture (I only do two bottles instead of three):

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In 7 days, you will split this culture.

Two new bottles should be filled half way with 1.019sg saltwater plus 1 ml Micro Algae Grow.

Pour your full bottle of phytoplankton through the rotifer screen (or a couple of coffee filters if you don't have that) into a clean container. Now pour that phyto into each bottle filling them both up to within ~1" from the top.

Those bottles are now ready to go into your station, starting the process all over again.

The other bottle you cultured should be run through the screen and then poured into a new clean bottle. This must be stored in the fridge and fed to your tank every other day. I pour about 1 cup in my 29g and 2 cups in my 55g.

It should last at least one month. I use up almost one bottle a week, so about once a month I have one bottle too much, which I can give away (or accept a small donation for the grateful reefer).

You need to shake the stored bottle daily to avoid the cells settling to the bottom of the bottle. Otherwise they will smother the lower ones and die.

It is important to keep everything clean to avoid contamination, including the rigid tubing that pumps air into the 2-liter bottles. That is why I keep using new bottles (rinsed out coke bottles) rather than reusing the same bottle it was just cultured in. You can clean the rigid tubing with vinegar water (or rubbing alcohol), or simply replace it with new tubing once a month. If you reuse your tubing, check the end to make sure it hasn't become clogged.

Feel free to ask me if you have any more questions.]
 
[Absolutely. DT's costs at least $30 a large bottle, which I used up every month.

Setting this up cost me about $25, since I already had the light. The only cost is the 1/2 cup of salt I use every 2 weeks. The rest is recycled as new batches are grown.

I'm even going to use my old PC lights that have lost their spectrum, because I'm guessing it is the perfect spectrum for growing algae.]
 
[Thanks Marc! Great directions.... I'll have to order some items from FAF. Plus, I am trying to raise my clowns, so I need this to feed the rotifers. I'll probably go ahead and order some roti-rich also just in case I have a phyto crash. Thanks a million for the clear directions!!!]
 
[I'd love to hear how you start a rotifer culture. I've had my starter sample in the freezer for about a month, and I would like to at least try them once.]
 
[Marc,

I've always bought live rotifers. But I believe all you do is put the cysts in water with a flow of air bubbles from a straight tube (enough to keep a decent flow in the water...I'd say like phytoplankton). It takes them 24-72 hours to hatch. (I think I have that time right.) Then you just feed them phyto. I forget how often you should harvest some so you don't have a crash...depends on the tank size you are raising them in.]
 
[Marc,

Raising rotifers is easy. You can use a couple of the same bottles that you use for phyto. Use a slow air flow, maybe a bubble per second. Feed them a couple of inches of phyto per day once you get a good amount of rotifers built up, less at first. If your water still looks green then wait for it to clear before feeding again, but they need plenty of phyto to multiply. Make sure not to contaminate your phyto when feeding or you will crash your phyto.]
 
[Steve,

Can you provide the steps from starting from scratch perhaps? 2 liter bottle, rigid airtubing, and 1.019sg water, plus the starter rotifers? And 2" of phyto?

Do I pour some out every day to be able to pour phyto in? What I'm pouring out will contain some rotifers, right? I take it they will be near invisible to the naked eye.

Do you rinse them while they are in the screen? Or just lower that screen in the tank to swish them out?

It sounds like I'll have a food source daily. Should I feed that during lights out when the corals are extended perhaps?]
 
[Set up two bottles just as you would your phyto bottles, except at one bubble per second. For the first few days start with one inch of phyto in your rotifers every day. Pour out just enough to make room to add your phyto. If the water you pour out still has a green tint to it, then skip that day of adding phyto. In a few days you will be able to see more and more of them. Some people, like Flame Angel, do it differently. but this works for me. You can check out her site for other ideas on feeding.

They are easy to see as they build up. Before my crash I would pour off two cups every night, pour them through your rotifer net, and rinse them with tank water. I am not sure that this is necessary, but I do it anyway. Then refill with phyto. I feed after lights out.

Did I cover everything you wanted to know? Here is a picture of my setup. Phyto in the back, rotifers in the front. Opening a valve drains phyto into the rotifer cylinders. A valve at the bottom of the rotifer cylinders is used to harvest rotifers.

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[Do rotifers swim or sink? Do they act like brine shrimp, attracted to the light?

Each time you drain some fluid to make room for phtyo, do you harvest rotifers for feeding?

Why two bottles?

I know phyto needs lights on for 16 hours, but I thought rotifers only need 10 or 12 hours...?

Sorry about all the questions. If I'd just do this already, it would probably be much more clear.]
 
[No problem......ask whatever you want to know. They swim/get blown around by the current I guess. :rool eyes: Many of them will attach to the sides also. You can gently shake or stir the bottle before you harvest to get them all off of the sides. They will get fairly dense after a couple of weeks. Normally you would feed each time you harvest them, but you do not have to. They will go down the drain without a fight if you don't want to feed as much. :lookaround:

I use two bottles for a backup in case one was to crash. Mine are on a 16 hour schedule like my phyto.

I have found rotifers much easier to maintain than phyto.]
 
[@Marc wrote:
Susan mentioned that she'd stopped adding Micro Aglae Grow to her medium said:
Can you explain this to me? I am not clear on what you are doing. You are not using any fertilizer at all?]
Click to expand...
 
[I bet if we talked on the phone, this would be sooo much more simple to grasp on my part.

Let me type what I think you are saying and you can correct me if I've off base:

Each day, shake the bottle of rotifers to get them floating. Drain off 1" to 2" of the fliud (what salinity btw?). That fluid is poured through the rotifer screen and rinsed with tank water. Pour the rotifers into the tank.

Replace the missing water with Phyto.

What constitutes a crash? Pure sedimentation? What do you do to avoid it? Feeding daily is the key apparently.


To answer your question, Susan originally stated to mix 2ml M.A.G. to the 1.019 water, then pour that into the 2-liter bottles. She's amended that suggestion with putting 1.019 water into the bottle, then adding 1 ml of M.A.G. This increased the amount of fertilizer per bottle and has created more dense cultures.

If you'll not the picture above, once I did that my culture became dark green compared to the week before.

Let me know how your tank is doing btw. It's been over a month since I heard the bad news.]
 
[If you want to talk by phone we can do that, but it looks like you understand. That is up to you.

Do not shake the bottle, more like just swirl it gently to get the rotifers off of the sides before you harvest them. Too much movement will damage them, hence the very slow bubble rate. Do this only when you harvest, and once you get a dense population you will not even have to do it then. Use the same water as you do for your phyto.

A crash usually means a large die-off, but I have never lost all of them. Drain off 3/4 of the water, keeping what is on the bottom out of the pour-off water, then clean and refill with the new water, and they will crank up again quickly. It is no big deal like a phyto crash can be. I have no idea what causes it, and it is not common, but will happen once in a while.

Remember, when you drain off the water, if it is still green you can hold off for another day before feeding again. Depending on the size of your rotifer population and the density of your phyto, feeding amounts can drastically vary. It is not an exact science. :cool grin: Go for it!]
 
[I'll start up the culture tomorrow I guess. I've got everything I need to do it.]
 
[I started my Rotifer culture yesterday evening.

According to the instructions, I soaked the seeds (embryo?) in a petri dish with 1.009 sg for 24 hours, with the cover in place.

Tonight I added about 2ml of phytoplankton. Replaced cover.

Tomorrow night, I'll do the same.

Finally on Sunday night, I'm to transfer my starter culture to one pint of saltwater and a little phyto.

To be continued.....]
 
[Okay Rick, I need more input. I've had my rotifers going for a few days now, as posted previously.

From the instructions, it sounded like the phyto would be consumed in a few hours, but it doesn't seem to be happening that fast. At least, not yet.

I've had them in a 2 liter bottle since Sunday.

The issue I'm running into is that the bottle is full and I don't have room to add more phtyo (food). So if I pour some out, I lose rotifers. If I screen the fluid, I have some rotifers in the screen, right? How do I get them into the tank?

Do I rinse the screen in the tank, shaking them out into the water that way?

Do you do this routine every day?

When do you feed rotifers? I would imagine after lights out.

Do you stop the skimmer so the rotifers remain in the water column longer?

Do I ever split the culture or just get a little bit from it each day? Does the fluid ever need replacing 100%?

The instructions pointed out to hatch the rotifers in 1.009 sg, but my phyto is 1.019sg. That is quite a jump. Any issues with steadily increasing salinity?]
 
[I shouldn't even respond for calling me Rick, but I will let you pass this time. :laugh long:

As I stated before, the rate of consumption depends on the amount of roftifers you have and the amount of phyto you feed. If the water still has a green tint then you do not need to feed any more yet. Wait until it is almost clear. Do you see some rotifers swimming in the bottle, or is the water too green to see into?

I feed rotifers to the tank after lights out when the corals are all fully polyped out. Yes, dip the screen into the tank to wash them out. I feed almost every night. I leave my skimmer running, but turning it off isn't a bad idea. I didn't figure it trapped enough of them to matter.

When the inside of the bottle starts to get a lot of algae buildup and dead rotifers of the bottom I (without shaking) pour off about 75% of the water and clean the bottle. I do that about once every 4-6 weeks. I have never kept mine in anything but 1.019-20. Maybe .009 is better, I don't know. But with the constant feeding of phyto at .020 being added, it would soon raise the level to that point. That is how I see it anyway. :glare:]
 
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