Culturing Isocrysis

[mwp]

OK...so...3 attempts with FAF Disks, and 3 utter failures. Nannochloropsis, Nannochloris and Tetraselmis are all going gangbusters EVEN AFTER some "abuse" during our move. But for some reason I can't seem to get an Isocrysis culture to take foot. Our setup is perhaps a bit "overlight" as I utilize an orchid growing rack for our cultures as well. Basically, there are four 48" bulbs (basically daylights/ cool whites) overhead, ranging from 18 to 30" away...these are the orchid lights. Laying on it's side, right next to the 2 liter pop bottle culturing vessels, is another 48" shop light with 2 bulbs (Coralife Actinic & Coralife 10000k).

Here's how it went down:'

1st try. Disk into 2 liters sterilized f/2 water under the 4 light fixture.

2nd try. Disk into 1 liter sterilized f/2 water under the 4 light fixture

3rd try. Disk into 1 liter sterilized f/2 water under the 4 light fixture + 2 light actinic/10k supplementation.

I have one disk left to try. All of my algae cultures have been thriving at 1.023-1.025 sg mixed with dechlorinated tap water, room temperature + heat from lights (so, 70F+). I've changed lighting, changed other stuff, and the only other thing I can try at this point is mixing it up with distilled water instead of tap. Any suggestions? Anyone having good luck with Isocrysis?

Thanks!

MP

 

[clownfish75]

what for of iso is it? Tahitian iso is ment ot be better in warm waters 25oC+ normal iso is fairly cool i am lead to believe.

Christian

 

[Luis A M]

Your set up looks fine.Some suggestions:
Get very good cultures.Might be expensive but you only pay once.
Use 1.010 ASW,(tap is OK) with GuillardÃ"šÃ‚´s.Chlorinate for 24 hrs.
Use only enough water so as to make it light green(or brown).When it starts to get darker,you fill it up.
Good luck!

 

[Kathy55g]

isochrysis. like also crashes.

 

[mwp]

Christian, honestly I'm not so sure what "form" - it's simply sold by Florida Aqua Farms as Isocrysis.

So Luis, the start disks are running like $8-12 a pop - not cheap, and from a reputable source. They've suggested starting with 500 ML, I've been using 1000-2000 ML. Even at 1000 ML, it comes out brown for the first day or two, but then the stuff dies off. Do you think a lower Specific Gravity, as low as 1.010, might be the ticket?

MP

 

[Luis A M]

<a href=showthread.php?s=&postid=7010808#post7010808 target=_blank>Originally posted</a> by Kathy55g
isochrysis. like also crashes.

and in spanish it sounds :"hizo crisis"

 

[FuEl]

Think clownfish75 is concerned with whether it is Isochrysis galbana or Isochrysis clone T-ISO. The former is best cultured around the 20's to 25 degrees celcius at max. The latter strain can tolerate salinity ranges anywhere between 7ppt-35ppt and temperatures anywhere between 15-30 degrees celcius. Due to this fact most people culture this strain. As your conditions you provide seem to suit both species, I'm stumped where the problem lies.

Are you using the same culture medium you use to culture Isochrysis compared to your other microalgae cultures? If so the media might be lacking in silicates which is crucial for survival for Isochrysis. You could try growing them at 24 hr photoperiod to hasten their growth rate (if you are currently using only at 12 or 14 h). Then again it problem might lie with the start disks, maybe try obtaining a stable culture from someone and trying it out?

 

[mwp]

Afternoon FuEL - I'm guessing that I have regular old "Isocrysis galbana". I am using the same culture media as all the rest (Florida Aquafarms "Micro Algae Grow", basically f/2 Gulliards). Silicates...hmm, that would in some respects make sense. Artificial Saltwater mixed up with tap tests out pretty low in Silicates. Our photoperiod is around 15 - 17 hours - basically on around 9:00 AM and off around midnight, 1 AM, or slightly later.

I've wondered about the culture disks...unlike all the other disks I've used to start, the Iso disks come off in flakes vs. simply rubbing off into a green liquid. I've upped the soaking time on #2 and #3 and this did not improve the "flaking".

Anyone know where I can get some active cultures? Just as FuEL suggested, this is the next plan if the last disk fails.

Thanks!

Matt

 

[FuEl]

Hmm..think I'm wrong about the silicates thing. Brain somehow got mixed up between Chaetoceros and Iso. Never started cultures from culture disks before but since you mentioned they were flaking off, it does'nt sound like a good thing. The problem should lie with the disks.

 

[GreshamH]

Nope, your right about the silica thing

Ask some of the major suppliers of phyto if they can supply you with a live culture

 

[TamiW]

mwp, I see you're in chicago. A few stores there carry Phyto2, which is pure strains of phytos, and one is isochrysis. Unlike when DT's started mixing their strains, you can culture this. I know that Coral Reef sells it, as does Reef Dweller.

 

[olin]

I've had the same experiences with iso disks. If I need to start a new culture, I use the 25ml liquid cultures from FAF. I do use silicate in these cultures too. Iso seems to be really picky in starting. To get them sucessfully off of plates, I've had good luck with starting with 3/4 strength aged seawater from a tank (microwave sterilized or autoclaved), and letting them sit and soak in the petri dish under cool-white fluorescents for 2 days. Then I move them to 250ml with fertilizer at f/4 strength (250ul/L). It works usually, but the liquid cultures are much easier.

 

[GreshamH]

How long do you microwave stuff for? Does it have to come to a boil in the microwave, or will the microwaves kill the living cells with out a boil?

 

[olin]

Usually 8-10 minutes for 2 liters of water. It will boil, but it's the microwaves that do most of the killing, causing DNA damage and structural damage that kills better than boiling alone. It won't kill everything like autoclaving, but it will clear out cilliates, vibrio and most of the nasties for aquariums and cultures. Pretty much only encysted bacteria may survive, but often these are completely cleared as well. I check sterility pretty often on nutrient media and it is rare to even have a single colony form.

 

[spawner]

Why does every think they need to use disk to start a culutre, this one of the worst ways to try to start a culture. Most plates are very containmated esp. those that are solid algae. The proper use for plates is to isolate strains, so you streak them as you would do bacteria cultures, not have a uniform plate of algae. These are not helpful at all and I have always wondered where this very bad mis information came from. The only thing people should be using plates for is to get very specific strains of cultures that is it NOT STARTERS!!!!!

If you want to get a good starter culture you need to get a clean starter in the form of a ultra pure test tube or a clean culture. The best way is to get a test tube with about 20 mL of culture in it that came directly from someones proven stock cultures. These are the cleanest cultures, next to that you should get cultures that were one scale up from stock cultures.

I would write more, but I am remodeling the entire condo right now.

 

[Atticus]

The plates are used because of the long storage time. I too have have pretty large issues with the Iso plates from FAF. Same flakiness and inability to thrive. I am very interested in the silicates theory as I use RO/DI mixed with IO salt and microwaved. I have tried the culture in 1 liters and 500mL containers with no luck.

 

[Luis A M]

I fully agree with the need of a good quality starter culture,though Atticus is also right in that plates provide a low maintenance storing mean.
I donÃ"šÃ‚´t want to say FAF disks were bad,many people have started algal cultures with them and theyÃ"šÃ‚´re still in the market. But they were not species reliable,sometimes you got different species than what you expected.And I think their ISO is not T-ISO.As I say,for starting and keeping an algal line,you must use a proven,preferably sterile culture.

As for silicates,they are needed only for diatoms.

Regarding microwave sterilizing,you donÃ"šÃ‚´t have to boil the medium.Several small capped culture vials are placed in the oven together with a 1-2 quarts pyrex full of tap water.Then run it at high following this program,given to me by Gary Wikfors several years ago:

3Ã"šÃ‚´on
5Ã"šÃ‚´rest
3 Ã"šÃ‚´on
5Ã"šÃ‚´rest
4 minutes on.

For larger volumes,chlorine sterilizing is more practical though.

 

[mwp]

Thanks for all that great input! It's not that I necessarily WANT to start with disks, but rather that it's all I've really been able to find out there online FishGrrl, I'll definitely look into those local shops and see what I turn up, and I think by this point, if I end up with 4 "false starts" on those Iso disks, maybe FAF would be willing to consider my investment and shoot off a small starter culture in a vial?

Wish me luck, I went with a reduced SG and distilled this time around and will soak it for 48 hours vs. 24+, and THEN will likely go real small on my initial culture volume. Something has to work

Matt P.

 

[ezhoops]

Iso sucks, good luck though.

 

[mwp]

Well, I let the disk soak for almost a week...didn't help much in the "flaking" department, but it didn't die on the disk either. I set it up last night with 500 ML of f/2 water made with distilled water. 8 hours later it still doesn't look very promising..pretty cleared out

MP