Culture Journal, Species: Apocyclops panamensis

General
Species:  Apocyclops panamensis
Species description:  egg carrying cyclopoid
Culture source (link if possible):  Kind gift of Jim Welsh, arrived 8/12/2011 via Fedex, room temp.
If algae, CCMP # (Optional): 
http://ccmp.bigelow.edu/
Culture Establishment Date:  8/12/2011
Continuation Date:  11/12/2011

Culturing Vessel Details
Salinity:  20 ppt
Temperature:    75
pH:  8.2

Vessel description:  One gallon square jar with wide mouth lid, hole in center, rigid airline connected to pump
Lighting description:  5500K fluorescent bulb,
Lighting cycle:  14 day/ 10 night
Aeration description:  rapid, but not boiling

Methodologies
Split methodology: Not sure yet, just trying to get a dense culture.  Weekly, decant culture to clean jar.

Culture medium description: 
Food is 6 parts N-Rich, 1 part Roti-Grow+ (6NR/1RG+).  6 drops per feeding, 2 feedings per day.  Equal volume of CloramX at each feeding.    
Starting at week 2, 1/2 cup of O. marina culture added per day as well.     

Cell count: Not known.
 (if known)

Reference links:  

Additional Information
(No Pictures or Videos in the Section Please)
Notes: 



You will be required to provide photographic evidence and as much detail as possible about your project in this thread.
If your thread does not contain detailed enough photos  and information the MBI Council will not be able to approve your reports.

Today I did the first jar change. I decanted the culture into a clean jar and suspended the sediment and poured that into a couple of tall tubes.  I let that settle and took a video of the little twitchers:

Here's some pics through the microscope:

 

after the tubes' sediment settled, and after I took video and pictures, I decanted the pods back into the culture, reserving the sediment to the trash.  
 
I am really suprised at the density of this culture.  What I thought was detritis, was really pods in suspension.  
 
There are significant numbers of ciliates in this culture, but they appear to do no harm.

Looking good, Kathy!  I see lots of gravid females in the video.  Was the video taken today?
 

Yes, it was taken the day it was posted, which is yesterday, now. I put some sediment in one of the tubes you've sent me, and then took the video directly from that after it settled.  This culture has done remarkably well, so far.  Let's see if I can keep from crashing it.

Jim, do you have ciliates in your culture? Or did I acquire them here?
When will we be able to get Culture points for this species? I am surprised it is not already accepted at the MBI!

So, the fact that you have nice gravid females means your feeding protocol is working well!  I probably do have ciliates in my culture.  I've submitted a classification request for this species, but so far, it has not been approved yet.

I was concerned about the amount of detritus in this culture, so I turned the air off for a couple of hours to get it all to settle.   The copepods continue to swim in the water body.
 
I decatanted the square jar and got a nice clean culture from it.  Square jars are easy to decant, because one can just tip it into the new clean jar, until it is sitting on its nice flat side. The detritus stays in, and the majority of the culture is in the new jar.

Here's the culture shelf:

 

Now there are strands of detritus in the jar. It's Friday, and I'll decant again on the weekend.  Culture looks cloudy, and i wonder if there are bacteria in it.  Could it be the O marina? 

Decant some off into a clear glass container (I would use something like a 250 ml Erlenmeyer flask) and let it sit for a few minutes.  Shine a flashlight diagonally from the back (flashlight at 2 or 3 o'clock, with your eyes at 6 o'clock, if that makes sense) and use a magnifying glass to examine it.  If your magnifying glass is strong enough, the O. marina will be little specks or "sparkles" swimming around.  Bacteria will just be cloudy still.  It also could just be tons of nauplii.  A dense culture can look cloudy when bubbling.

I took a pudding cup's worth of culture, put it thru a 37 micron, back washed into a petrie dish (6 cm diameter):  CLICK on the video

Thanks, Jim, I'll try it tomorrow.

Jim, that 2 oclock, 6 oclock light angle is brilliant!  It really helps to see the culture.
 
A. panamensis cultures are doing well, better than anything besides O. marina.  They have outcompeted T. californicus in the polyglot bucket, as well as doing pretty well in the simple culture with O. marina.  I have O. marina everywhere, and I almost have A. panamensis everywhere.  Not sure which culture is more successful, the one that I fuss over, or the bucket on the floor that gets all the leftover pods and rots, with just feeding and little or no water change.
 

After a couple of days of starving the culture during MACNA, I've discovered that precious few A. panamensis are still alive in the single culture, while they are still thriving in the polyglot bucket on the floor, and I haven't looked under the microscope, but there seems to be a healthy population outside as well.  I've sieved them out of the water of the single culture, and given them clean saltwater, a dose of O. marina, and some of Andy's recipe phyto.  I hope they bounce back.

They are not bouncing, though they appear to still be there. I feed them sparsely, and hope.
The outdoor culture, that was just seeded with rotifers and A. panamensis is really doing well. It's 62 degrees this morning and a quick dipper of the culture, filtered thru a 37 micron and back washed into a petrie dish revealed several adult A. panamensis, very few rots, and 20 times as many star shaped nauplii. That was about 1/2 cup from a 17 gallon tub outdoors.  Interstingly, last week that same tub had a dense culture of rotifers as well as panamensis.  I guess the natural algae population could not keep up with the rotifer explosion. A. panamensis is sturdy, indeed.  I may begin feeding this outdoor culture just to ensure that it continues.  I may start an indoor culture from it, as my "clean" culture still struggles to proliferate.

Here's a vid of a microscope view of the seived dipper ful of outdoor culture: Click on it!

i took a larger sampling of the outdoor culture, and it has a lot lot lot of M. salina adults in it.  I'll have to check the nauplii appearance of the M. salina and the A. panamensis.  I may have been mistaken, actually it is likely that I was. 
 
My indoor culture has bloomed, and I can now start increasing the volume and feeding more O. marina.  Yay!

I think there are still A panamensis in the outdoor culture, though there are no more rotifers in it, and there are definately some M. salina in there, splashed over, no doubt, from the neighboring tub.
 
The A. panamensis indoor "clean culture" are slowly increasing in population. I continue with a few drops of Andy's mix and chloramX  twice a day, and about 100 ml per day of O. marina culture.  Weekly decant to a clean gallon jar.  The culture remains cloudy green.  Sometimes I use RGcomplete instead of the Andy's mix and ChloramX.

Nearly crashed it.  It turns out the O. marina had gone south, and I was adding ammonia water and ciliates to the culture of panamensis each day.  There were enough to restart, however, and I have a saved jar of panamensis from my outdoor culture when it bloomed a couple of weeks ago, as a backup.  
 
Gotta love the great outdoors.  I've noticed that populations bloom in a big way, and then crash out there, making way for the next bloom of something else.  It's pretty interesting and fun.  Next year, I'll document better what I do and what the results are.  This year was just playing around.  I am surprised it was so prolific given that i've done next to nothing to keep it going.
 
I sieved the jar of panamensis through a 37 micron filter, and put the pods in some clean water with RGcomplete.  This evening I fed a few drops of Andy's mix.
 
I've filtered the O. marina as well, keeping the flow through, and hopefully the ciliates contamination did not kill everything.

The jar of panamensis recovered from the outdoor bloom are doing well with just a little food and no additional air.  The indoor ones with daily food additions, just RotiGrow, NRich and ChloramX , and air and water additions are doing OK, though not as dense as they had previously been.  I changed the jar today.
 
Still concerned that my O marina crashed.  I haven't been adding any O marina from my culture to the copepod cultures.  I kept a jar of something that I got by filtering one of my other cultures (flow thru) and I think its O marina.  I added rotifers as a size reference.  What do you think? Are the dots O. marina or ciliates? 

I have a couple of cultures of O marina. I'd be willing to give you one of them. well I guess more like give you one back lol.

RLTW

180 Gallon Mixed Reef

Then I heard the voice of the Lord saying, "Whom shall I send? And who will go for us?" And I said, "Here am I. Send me!" Isaiah 6:8

Just a sample would be great. Call me when you're going to be in town.

will do.

RLTW

180 Gallon Mixed Reef

Then I heard the voice of the Lord saying, "Whom shall I send? And who will go for us?" And I said, "Here am I. Send me!" Isaiah 6:8

Seems to be doing better lately, seems quite dense.  I'll have to split today so they don't crash.  Perhaps I'll start a 5 gallon bucket now.
 

wrong again. they were not too dense, and a lot of rotifers in there.  I filtered and changed to a clean jar and clean water.

Here are videos taken this morning.  The first is a shot thru the culture with the help of a flashlight.  The second is 40x from the microscope. There are copepodites and fast swimming parental pods.

 

I kept the filtrate from one of my cultures, can't remember the filter size, darn it, and set it aside at room temperature. Now have a culture with lots of juvenile A. panamensis!!!  I may be able to have a pure culture afterall.  Let's hope there's no rotifers in it, as I have been keeping them near the rotifer buckets.
 
 

I'd be worried about the Apocyclops contaminating my rotifers!
 
I find that some, but very few, Apocyclops nauplii will pass through an 80 micron sieve.  Many, many of them will pas through 120 microns.

It's been a while since I logged in here. I've been ignoring my shelf cultures, but I have some panamensis in my S rotifer culture.  I'm thinking its a good thing.  The rotifer culture is cleaner and the panamensis get fed regularly. The panamensis are thriving, and there seems to be little deliterious effect on the rotifer population.  I want panamensis to feed to my larvae, and all the larvae get rotifers, so why not culture them together?  I wonder if this would work with parvocalanus and other desirable copepods.
 

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

I've also got panamensis and S rotifers in a tub outdoors, and they are thriving.  I first seeded the tub with some green micro algae, the remains of my isochrysis attempt, probably chlorella. After a couple of weeks of consistent green, I added a couple of gallons of the S rotifers mixed with some panamensis. Today, a couple of weeks later,  I put 4 ounces thru a 53 micron filter and backwashed into a petrie dish.  There was a good culture of rotifers, and hundreds of panamensis nauplii, and lots of egg bearing females.  They are doing really well outdoors.  High temps for the last week have been in the low 90s F.  It's quite possible that the panamensis are eating the rotifers….

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

Eating the rotifers?  That's interesting...  Aren't the panamensis smaller than S rotifers?

Don't let fear and common sense stop you! =]

I doubt the copepods are eating the rots. They are filter feeders and only eat really small stuff. It's quite possible to me that you've had a bloom of some small protist and the copepods are eating that.

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

I'm with Andy on this one.

After the heat of last summer, all my outdoor cultures crashed.  And with enough neglect, I got some of the A panamensis away from the rotifers in my indoor culture. Now I have a great culture of A. panamensis, in gallon jars, fed only with live Iso.  It's amazing.  The cultures are pretty dense. I feed them very lightly with Iso, maybe 100 ml when they turn clear. Every week I change 100% of their water and give them new jars. I keep getting more and more cyclopods! I'll get pix in a short while.

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

 
Isn't it, though?

Freaky thing: Rotifers crashed just as a small nest of ocellaris hatched.  I've started counting the A. panamensis I'm culturing as I almost lost them by trying to concentrate them too much. I had 6000 nauplii leftover after seeding my jars, so I gave them to the 6 gallon ocellaris larval tank along with what few rotifers I had. An hour later the larvae had noticable bulging tummies, and it's been like that for 3 days.  I've made no further additions aside from the rotigreen I added on day one for green water.  I'm going to get some more rotifers tomorrow, as I am afraid the remaining copepods will grow to be too large for the larvae before the larvae metamorphosize.  There's only about 12 larvae in there, but they are not dying, and they look active and healthy.  

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

Nice!

Well, I was thinking that feeding with A panamensis might be better than rotifers, judging by the success of the small nest of ocellaris mentioned above.  I had a larger nest hatch out, and I put them with the 4 day old smaller batch, and thought they were doing well.  I added a lot more panamensis nauplii, but did not have the same response as the previous batch. In fact, many of them died over the 3 day period, as well as some of the previous dozen or so.  The ones that survived look good. So this is not a tried an true method, and I still have some work to do.
This week, I have 3 nests hatching within a day of each other, and plan to put them all in the same tank, so I will be feeding them rotifers and panamensis nauplii and I'm really hoping that they do well.  I kinda need these fish….

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