Phytoplankton

[jonboy]

Frank,
Thanks for the articles last month and this month about producing the green water and the rotifers.
I started out last month with the green water.
I started 2 seperate 2 liter bottles -1 with dt's ,2nd with algae from FAF
both bottles are set up the same ...the dt's is yellow/brown while the
algae is lime green.
I have divided the containers twice now.
I was wondering why the difference in color.

I have since divided the algae into a larger container ( the start of a
phyto reactor) it has turned yellow/brown also. Only 3 days since the
change.

Another observation...tonight it looks like one of the dt's is changing to a
green color.

I don't think that I'm color blind.
Have I done something wrong or is this part of the cycle?
Thanks
Jonathan

 

[FMarini]

Jon:
great question...Allow me to ask. Do you add fertilzer or F/2 to either of thee cultures and if so how much?
So my intial impression is that you had 3 things going on here. 1) a very low density culture from the DTs, and 2) possibly a deficiency, or 3) a contaminate.
So point 1, I have read a number of times from people that much of DT live phyoplankton is dead or let me rephase this not viable. In fact many people have reported to me that they cannot start cultures from this product. I have, and i only buy a fresh product which has been shiped and stored cold. neverthe less, if you start your culture w/ a very low starting density then your eye will play a trick on you. Very little phyto=light green or brownish(as in the figures in my article) as the culture achieved higher densities your eyes start seeing this as darker green. Now the algae discs from FAF are fairly high concentrated stuff, so i suspect you had an excellent innoculum. Second point, if you are adding the correct amount of F/2 or any home made fertilizer you should be fine. yellowness or brownness in a culture is indicative of iron deficiency. However, brownness can also mean contamination. A third possibliity is that you had a contaiminate in the DTs and that it took s awhile for the Nanno(which is bright green) to overgrow the culture. IME, whenever you culutre more than one type of anything together...eventually you always get a monoculture.

 

[jonboy]

Frank,
I have been using the f2 from FAF. almost 1ml per 2 liter
All the bottles are hooked up to the same air pump.
2 bottles of the dt's have turned brown one is green
2 bottles of the Nann. algae are brown one is green
I have used microwaved water for each of the additions.
In my large container that is brown now ,i used ro water.
I contacted Flame angel and she said that she uses ro as the
make up water.
I can attach a photo if you would like to see it.
I don't think that I contimatinated the algae but I may have.
I allowed the air pump to run while I changed the bottles and
refilled the water/f2.
At this point , would you wait and see what happens or start a new
culture. ????

My original culture was dark green before i divided it about 8 days.
The original dt's never turned green, was light brown but i divided it
anyways. I had read that dt's was a mix of algae's and that was the
reason that the bottles were different colors. I may have guesssed
wrong.

My main concern is with the large container. That has turned brown.
Ahould I lose that culture and start a new one? I hope to build a phyto
reactor and need a good starting point. If I start over, what should I do
to sterilize the vessel? It is 4'tall.
Thanks for any help,
jonboy

 

[Louis Z]

Hello jonboy, Hello Frank, I had this problem happen to me with a culture of Isochrysis and Tetraselmis. I received both from FAF. The cross contamination occurred due to the fact I used the same gangvalve from the same pump on both cultures. The first week everything looked great for I got extremely deep colored cultures- one brown and one green. Boy they were pretty to look at (translucent) - I was so proud. The second week after I split the culture I noticed that my Iso was tinted green. Well my Iso culture now takes 12 to 14 days to show after the tetraselmis has degraded instead of the 7-10days. I still culture this but I will eventually reorder the culture. I also see alot of cloudiness to both cultures so I realize my non -sterilization technique was a major contribution to this. I knew that I wouldnt be able to keep it bacteria free forever but I didnt follow the Plankton Culture Manual as I should have (rookie mistake). I will now try to keep continous small sterile batch cultures going in the small vials as the ones I received from FAF. I have treated my cultures as Rodney Dangerfield says "I get no respect". So I have to repurchase due to my inexperience and laziness. As for the 4' reactor - How does one go about cleaning it. I do notice a lot of scale buildup (I assume from my poor Ph maintenance) on the 2 liters and the rigid tubing. Hopefully my new CO2 dosing rig on my 2 liters works so as to avoid the Ph shifts. Thanks Louis Z.

 

[Louis Z]

I forgot to tell you about the sterilization I am about to do. I got tired of Microwaving small amounts of water. It would take me close to an hour to do all what I needed. So I went and got some pool chlorine powder (calcium hypochlorite) to use as a water and equipment sterilizer. johnboy I think that Plankton Culture Manual is very important to have. It discusses sterilization techniques and helps problem solve in culturing.

 

[jonboy]

LouisZ,
Thanks for the reply. I will get mycopy of PCM out and reread the info.

After your reply, I probably have cross contaimanation also.
Using un sterilized,but ro water may be the cause. However not all of
the bottles have turned brown.
And one of the bottles that was brown has turned green. would that
mean I have a longer cycle time also?
When you sterilize now, do you use ro water and then add the pool
chlorine? let stand 24 hours and then it is ready for use.

would excess temp cause a culture to crash? I may not have let the
microwaved water cool long enough...

Thanks for all the help,
Jonboy

 

[Louis Z]

jonboy I think the sterilization technique is more important with the continuous culturing of the innocculant (your PCM has the process). Even though you use RO there is psuedomonas that grows after the membrane so I assume anything else can too. The tubing from the RO to your vessel can also cause contamination. So all those points contribute to poor cultures. For the algae is what you want to take off faster than the bacterial growth. There is no way you can keep your reactors bacteria free but just allow the algae to take off first to reach maturity as quick as they can. So all those PITA steps are so tedious -I skipped them. Well I pay the price- now I am several months behind in what I wanted to do and must try to learn and practice technique with new cultures. I may never be able to duplicate what FAF does. I dont have a sterile room, nor sterilizing equipment so I will try to make do with what I have. It may be impossible for all it takes is a slip of the hand. As for using RO, I think that is better than tapwater for in the PCM it talks about unknown chemicals and concentrations as a possible hinderance. Yes your color changes do mean cross contamination and yes that may slow down your intended culture. As I said my Iso takes so much longer now its not really what I want to use after the Tet. has degraded in the same bottle. All this may not make much of a difference if your are feeding your tank but I want to do more than that (breed and raise fish). I do want to direct you toward a thread in Reefcentral that is just now starting to pick up with good advanced points about phyto culture(it s in the reef discussion forum.

 

[ADS]

Jonboy- It sounds like a contaminant or bacterial problem like LouisZ noted. I would dump the discolored cultures and split the healthy one. Try to keep at least 2-3 starter cultures at all times. Also check your specific gravity. I find cultures better at 1.020.

LouisZ-I have some experience w/ T-Iso and find that it is VERY temp sensitive >75 degrees. Cultures die rapidly and turn green. Nanno is much more Temp tolerant.

Adam

 

[Louis Z]

thanks Adam for the tip. Over this winter with my cultures in the garage they were subject to cooler temps. The Iso did seem invincible. I had the 2 liters in a 10gallon tank water bath with temp around 70- 72'F to keep them warm but also to minimize water evaporation out of the 10gallon.