Culture Journal, Species: Oxyrrhis marina

General
Species:  Oxyrrhis marina
Species description:  Heterotrophic dinoflagellate, ~ 15 microns wide x ~ 30 microns long
Culture source (link if possible):  Volunteer contaminate in my T-Iso culture
If algae, CCMP # (ref http://ccmp.bigelow.edu/ ):  CCMP 604, 605, 1739, 1788, 1795

Culturing Vessel Details
Salinity:  1.010 - 1.015
Temperature:  Ambient room temp; ~ 72 - 74 F
Vessel description:  500 ml Erlenmeyer flasks, 1000 ml Erlenmeyer flasks, 2 L media jars
Lighting description:  Ambient room light
Lighting cycle:  N/A - this species does not need light to grow.
Aeration description:  Varies; they do best with moderate aeration

Methodologies
Split methodology:   Harvest 1/4 to 1/2 of the volume of each container daily.  Split or feed to copepods as desired.

Culture medium description: 
8 drops / liter twice daily of varied preserved algal pastes.  Typical feeding for 1 liter would be 4 drops of Roti-Grow Plus (primarily Nannochloropsis) plus 4 drops of N-Rich (Primarily T-Iso, Pavlova, and other EPA-rich algae) twice daily.  Also add 1 drop of ClorAm-X solution (12 tsp. of ClorAm-X powder dissolved in 1 liter of water) per drop of food added at each feeding.

Cell count:
Very rough estimate is 50,000 - 100,000 cells / ml.

Reference links:  

http://sabella.mba.ac.uk/...ng_Oxyrrhis_marina.pdf

http://www.int-res.com/ar.cles/meps/2/m002p229.pdf

http://www.springerlink.c...tent/f204x254318278j0/

http://cat.inist.fr/?aMod...eN&cpsidt=16712370

http://www.int-res.com/ar.cles/ame/10/a010p307.pdf



Additional Information
Notes:  I first discovered this contaminate in my A. panamensis cultures in February, 2010.  I ultimately discovered that the O. marina had contaminated my T-Iso cultures.  I've been culturing the O. marina ever since.

 




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Attached Image(s)

Thanks for the information Jim
What magnification is that?

Doni Marie~

Can you snap some pics of the cultures in the Erlenmeyer flasks?
Thanks

The images were taken at a magnification of 100x, Doni Marie.

 

@Zooid, pictures to follow.  Gonna be boring, though!

LOL...that's ok....I just wanted to see the color.

When just fed, kinda yellowish.  When hungry, kinda milky whitish.  I'll post pics of both later.

Very nice pictures. Is that with phase contrast?

Some phase contrast, one using Nomarski lighting (whatever that is).  The microbiology folk at the lab I work at helped me with these.

Thanks Jim. I got my microscope this week and I'll need your assistance with confirming my cultures (both O. marina and the panamensis)   I THINK they are both doing really well. I am using 1.025 salt water for both with no apparent issues.  I did have to strain my O. marina with a 52 um a few times due to rotifers.

Doni Marie~

Doni: Any time I do anything with those cultures, I strain them through a rot screen. I've not had a lot of rot issues, though. The rots won't eat the O. marina and the O. marina hugely out-compete the rots when you get the food concentrations right. The bigger problem I've had has been with a large ciliate and with bacteria. But both of those also seem to be controllable with culture container changes and food supply control.
 
Jim: Yes, very nice photos! I need one of those scopes.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

Andy, with a hand held 10x magnifier, I can see a very dense quantity of uniform specks.  'Specks' is all I can say without looking at them under the microscope but they are much smaller than rots.
I am not familiar with ciliates and what they look like.
 
 

Doni Marie~

Sounds like them, though we won't know for sure without the scope. I can see the clouds resolve into little individual specks without magnification if I use backlighting. That's how I do my daily culture checks, in fact.
 
The ciliates I had problems with are much larger than O. marina, more on the order of slightly smaller than a rot. But get those slides in. We'll know for sure really quickly.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

Ok, I know the pictures are late, but here they are:

 

Before feeding:



 

After feeding:

Attached Image(s)

Thanks, Jim. I always wanted to see how you are culturing them.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

I wish I had the money for a few gallon sized Erlenmeyers hehe.
Thanks for the pics.  Looks similar to mine.  Andy's always seems denser than mine though

Here's a thought, Gale: What kind of lighting are yours under? My cultures that get the least lighting are consistently the best performers. Maybe the light encourages algae and bacterial growth?

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

I'm going to resurrect this journal.  I first noticed this organism as a contaminate in my copepod cultures in February, 2010.  I confirmed the tentative ID in March, 2010.  I've not been culturing these deliberately recently, so I'm just now re-"establishing" this culture for the purposes of this journal, and the MBI Culture Continuation report I'll be filing soon (after the 90 days have passed).  I know that I could probably file a report based on the success with this species that I've already documented, but want to demonstrate how easy it is to establish and maintain cultures of these little guys.  My intention is to act as an inspiration to others who may wish to culture them as live feeds for copepods and/or other creatures.
 
To "Establish" this culture, I took the water from a crashed Moina salina culture and examined a sample under the microscope.  I could see numerous O. marina, and also some Euplotes.  I filtered the crashed Moina culture water through a coffee filter into a clean flask.  I examined with the scope again, and there were still too many Euplotes for my liking, so I filtered again, this time through three coffee filters stacked.  After that, I could see no Euplotes in any of several samples I checked with the scope, but still had numerous O. marina.  I'm initially feeding them lightly with live Isochrysis, but will switch to using preserved algae pastes soon.

keep the info comming =)

Good luck, Jim!

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

I hate to resurrect old threads but I'm having trouble finding much info on these guys. Culturing seems simple enough, but aside from A panamensis, what other copepods is this ciliate useful for? I'm curious if its been tested as a food source for species with more potential such as P. crassirostris.

Nothing wrong with resurrecting old threads - great way to breath new life into a topic. I"m curious too.

I tried them with several starters of P. crassirostris from Reed. The adults seemed to do fine with the ciliate as a food source and certainly kept reproducing but the cultures slowly died off over roughly six weeks or so. My guess is that O. marina is too large to serve as a first food for the tiny baby copepods. I think that your cultures could do fine with a mixture of T-Iso and O. marina, but if you have to go through the trouble of culturing T-Iso in the first place then why bother with the mixture? Of course, using a mixture might mean that you could get by with less T-Iso and that might be a good thing.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

My thoughts exactly. It's the culturing of the Iso I'm looking to avoid (don't judge me!)

I almost tempted to purchase a starter just to see if I can get the naups through the early stages just long enough to be large enough to take O marina.

A tonsa is the other species I want to try O marina with.

It's easier to culture phytoplankton than you think. I mess with my cultures once a week, and I have plenty to keep 5 different copepods going and some of my rotifers as well. It's easier to grow the phyto, than it is to keep cultures of O. marina, and the pods do so well, in my opinion.

check out Kathy's Clowns, llc website:
http://kathysclowns.com
Captive bred clownfish and more
(Wholesale to the trade.)

It's not the trouble it's the space

 
I wish that had been my experience, Kathy. In all my tries with T-Iso, I've managed one split of one bottle total. Heck, just the cleaning alone. I kept O. marina for years in open-topped buckets.
 
Joe, the problem is that the copepods are going to have naups constantly, so you will have to constantly supplement O. marina if you want to keep the culture going.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886

Andy, you should try again. I find that there is some acclimation period where there is a pause in growth before it really does well.

check out Kathy's Clowns, llc website:
http://kathysclowns.com
Captive bred clownfish and more
(Wholesale to the trade.)

Actually was pretty successful with T Iso. I'm confident I'm able to culture phyto if needed I just honestly don't have the space for a culture station like I used to. I know everyone says that but if I give up the space I have for larvae I may as well not culture the copepods anyway lol

The O marina can be cultured in the space I currently have my rotifers, which I don't want to culture anymore anyway. That frees up enough space for something like A tonsa and doesn't encroach on my larval space.

I really should, Kathy. Thinking back on it, I think the only success I had was in the summer batch. I do wonder if I gave them too cold of an environment growing in the basement. The first thing I would try differently would be a warm water bath around the bags.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886