So, some of you should have cultures. How's it going?

Experiences. Questions. Comments.
 
How are things going?

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

So far, so good. I have several cultures of each going but haven't checked for density numbers yet. 

http://www.fishtalpropagations.com/#!home/mainPage
"Making captive breeding easier."

My O.marina is doing great. (I think... I ordered a scope TODAY so I'll know soon what I have.)
 
My pods started off in a culture of too much water (my fault) and they are off to a really slow start. I think they are starting to come around but only time will tell.

-Adam

Good, good. I'm glad hear it. (Mua-ha-ha. The evil laugh just seems appropriate for some reason.) Adam, they should ramp up really quickly. I've gone from maybe one or two individuals to them completely taking over a parvocalanus culture in a month. Don't give up on them.

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

My O.marina is hard to tell what's going. I need to reread the thread to identify what is it I'm looking for. Pods are still alive. What is the reason I can't feed then Rotigrow or Rotigreen?

They are really small, so you really need a back light to see them. Is your culture taking on a brown color, even when you are just about to feed again?
 
Pods: You can feed them anything you want to. They just may not survive on it. (See here: http://www.mbisite.org/Forums/tm.aspx?m=60961&high=.) I'm just giving you the method that works for me. Whether you follow it is up to you. Cultures fed pastes tend to ultimately be very unstable cultures. If you do that, I would keep a _lot_ of extra cultures running.

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

 
I see a bunch of little guys swimming around and then every time I check (daily) I see more and more adults. I dont have a scope yet (comes in a couple of days) to see how many gravid females I have but even if I did, I wouldn't disturb the culture right now.  I'll keep you updated.

-Adam

Everyday I look twice and it seems there are more and more large adults along with more little guys. I would say I started with ~20-40 adults and I think I have well over 100 now.
 
I get my scope today in the mail so I'll be taking some shots if I have some free time.

-Adam

Got my scope... @ 40x
 

 
http://www.youtube.com/watch?v=ORdEzMy5Q9w
 
@400x.. Man are these things fast!
 
https://lh5.googleusercon...RGr0/s800/Img00071.jpg

-Adam

Good looking culture, Adam!

Yay! You got your 'scope working!  

http://www.fishtalpropagations.com/#!home/mainPage
"Making captive breeding easier."

Possibly a molt?? I have no idea. Didnt move and no internal color.  Dead or a molt I think.
 

 

 

 
 
...and then just for fun.
 

-Adam

Molt.  That's also what my avatar image is, a molt of an Apocyclops copepodite.

Yes, very nice! Congratulations on what certainly look like some healthy dino cultures.

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

My cultures were taking a dive as it is getting cold in my basement. I just fixed that with a tub of water and a heater with my cultures in that and then it is covered. Hoping it will get my through the winter.
 
Ciliate?
 

-Adam

That looks a lot like O. marina to me. Without seeing it move it's hard to tell for sure. But I'd say flagellate.

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

@100x
 
http://www.youtube.com/watch?v=UQsZ70XILng
 
You can see the dinos all around this alien dino.

-Adam

Thanks. Seeing the movement and size differential makes it much easier. Yeah, that's a ciliate, and quite a large one. Maybe Euplotes? I'm not very good at protist ID.

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

I don't know what it is, but it has neither the right shape nor the right movement for what I understand Euplotes to look like.

Well, you have a lot more experience with Euplotes than I do, Jim. Yeah, whatever it is can be easily filtered out of the culture. But maybe let it multiply a bit first, then filter it and wash it into its own culture and see what you come up with? You might have the next big thing there.

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

I find it is hard to determine how well my O marina cultures are doing.  A bright light through a jar of the culture does resolve into individual specs, but they never seem to take on a brown color between feedings.  They get the lightest of green tint to the water, but the O marina seem to be white. Maybe I'm feeding too much, but I don't see detritus or uneaten algae settling to the bottom.  I'm feeding about 5-7 drops of RGComplete to a gallon of culture twice a day.
 
The A panamensis culture appears to be doing fine.  It's been a week, and while I haven't seen a decline in pods, I haven't seen a dramatic increase in density either.  I'm currently chalking that up to waiting 48 hours for the O marina cultures to beef up before I started feeding.
 
I haven't started a culture journal for either since my club is currently going through the process of becoming an MBI portal site and the journal board on our forum only just got started.  It's on my to do list though.

- David Gibson
Denton, TX

They'll be white when you are shining a flashlight through them (it's reflected light). The brown that I see is in a white bucket under room lights. Part of it is likely the food that I'm feeding (it's mainly brown).
 
But, 5-7 drops of RGComplete per gallon is not much for them. They eat a lot. I'm feeding 7 drops of food per gallon twice per day, and my food is three times more concentrated. So once your culture gets up to speed, you should be aiming for 21 drops (a little more than 1 mL) of food, twice per day. I would up the food quite a bit.

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

Ah.  I'll do that then.  I keep forgetting that the RGComplete is only a third the density of RG+.  Means I'm underfeeding my rotifers too.

- David Gibson
Denton, TX

In my case, I'm on my third try, this time in a five gallon bucket.  First two cultures just died off using the 2 liter bottle method Jim uses.  I'm using RG Complete, but going by the water tint and not a specific amount (might be my issue).  My first two attempts lasted about two weeks, actually split the second culture and both crashed.  The first just seems to have withered away, I was feeding regularly and the green would dissipate mostly and feed again, then the green stayed the same tint and I found no live copepods.  I'm starting to suspect my source since I got the from another (almost) local reefer and not a "normal" source.
 
My rotifers have also gone down hill and I'm going to restart them.  But this has happened before to me.
 
Jeff

Jeff, what cultures are you talking about failing? Copepods? O. marina? Both?

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

 
I have the same questions, Jeff.  Also, the only thing I remember saying I cultured using 2 liter bottles was my phytoplankton, so I'm confused.  Can you refresh my memory about what method of mine you are referring to?  It is hard to tell from your post what cultures are crashing on you.

My copepods are doing great.... BUT
 
My O.Marina are all dead. I think I let the cultures go too long between the times I siphoned "1/5 or more" out of each culture.
 
I'll have to hit up one of the locals for some more O.Marina.  *Sigh*

-Adam

Sorry.  Actually both.  O.marina was what I originally started the topic on and then strayed. 
Jeff

I don't even understand what I wrote. 
 
Here's the basics on my o. marina:
 
I received a culture locally, put it into a 2 gallon bucket.  Fed RGComplete and lost the culture after about four days (no more living cloud swimming around).  No real idea what happened, never even got the bucket filled and saw no life.
 
Got a second starter culture, put it in two buckets, added water for about a week until the buckets were full.  Fed the same, still had the swimming cloud and recognized larger particles swimming in one bucket.  Ran it through a 53 micron sieve.  Started siphoning out 1/5 a day and putting it into my refugium on the display tank (better than nothing) and got a bacteria cloud on one bucket (really cloudy, no specs).  Dumped that bucket, split the first bucket into two and started adding water again.  Within a few days there was no life in either bucket, similar to the very first run.
 
In the mean time I started copepods and was just ready to feed the o. marina when the o. marina crashed.  As a result, I'm pretty sure the pods just starved.  My rotifers crashed about the same time, but that's not an unusual event for me.  Right now I'm wondering if I contaminated the o. marina culture when I started copepods.  Only I saw nothing in the water sample, no copepods, rotifers, o. marina, or anything alive.
 
I need to get a 27 micron sieve to follow the rest of Andy's maintenance schedule, and I'm going to move the cultures to opposite sides of the room.  When I first started rotifers I went through 3 or 4 cycles before I had a handle on them so I'm not thinking this is any different.  I have switched to algae pastes and/or Rotigrow and that's solved some of my issues with roitifers, hoping I don't have to culture phyto again (phyto is what I learned from Jim's bottles...) for o. marina.
 
I may not set this up again until after Thanksgiving, but I'll start again.  Thanks for all the help, I may be slow but I'll get there. 
 
Jeff

Adam-- Well, if you're copepods are doing great then you don't really need the O. marina. Rejoice! I still have not managed to do without, though. I haven't tried very hard lately, I have to admit. The O. marina eats tons and water quality can go downhill fast. The water changes are pretty darn necessary.
 
Jeff-- Sorry, man. Tell us more about what happened and we can try to diagnose it for you.

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

Of my two cultures of A. panamensis, the one in the black bucket is doing quite well, the other one isn't. The O. marina in the shallow tray did better that the deeper one or the flask.

http://www.fishtalpropagations.com/#!home/mainPage
"Making captive breeding easier."

 
I think this attitude is a VERY important one to have when working with new cultures!  I've had to attempt almost every copepod species at least twice before I was able to be successful at them (and that includes Tigriopus!).