Adventures with Artificial Incubation for Banggai Cardinal Eggs

Well, I officially gave up on my male Banggai after he swallowed his third batch on just mere day 2.  Just yesterday I came across someone mentioning nitrate in the 10-20 ppm range can cause this issue, so I'm going to research this more as my broodstock system is usually around the 20-30 ppm mark.   In the meantime, I went to the fish room about 7 pm lastnight to see the male Bangaii holding some eggs.  He doesn't even seem to make the first night very well without consuming (or spitting?) the eggs, so after his last failed attempt I planned to strip his next batch as soon as I found him holding.
 
So lastnight around 8 pm I collected approximately 38 eggs.  I put the eggs into a San Francisco Bay pop bottle brine shrimp hatchery with approximately 600-750 mL of water from one of my reef aquariums (1.026 and nitrate undetectable).  I filtered through a 53 micron sieve, sterilized it with bleach, and dechlorinated.  I turned the air down fairly low - just enough to keep the eggs from settling on the bottom - and put a lid on it.  I also added one drop of methylene blue dye in hopes of it preventing egg fungus.  I then set the "tumbler" into the sump of my broodstock system to keep the temperature steady right near 80F (26C).  I plan to do 50% waterchanges every few days, and keep MB in the water.
 
Now, this is far from high tech, but I want to keep this simple if possible, so I'm starting with the simplest setup I think might work.  I have antibiotics and formalin on hand to try in the future if the MB proves to be unsuccessful.  I will take this project as far as needed until I achieve consistent and reproducible success.
 
I actually purchased 4 small adult Banggai Cardinals a few days ago in hopes to get at least one more pair going.  I'm going to try the "vent method" for sexing them once I can get their bellies full.  Right now they are just being weaned onto frozen foods and then I will medicate them with PraziPro for a week.
 
Wish me luck!  Feel free to make some suggestions, although at this point I suppose "waiting to see what happens" is probably the only suggestion because if this works I'm stickin to it!  Haha!  It seems there aren't many people successful with AI for Banggai Cardinals, and I've only had moderate success in the past.

In the net the eggs didn't stick together




In the tumbler the eggs stick together

 
 
This is how they look this morning, I'm assuming the blue ones aren't fertile?

Pretty picture.  I'm thinking that the blue ones are fertile, as there seem to be eyes…not sure.

Kathy, that's what I thought at first too, but then I was thinking they shouldn't have eyes yet anyway since Bangaii eggs are so slow to develop.  I read somewhere that MB will cause dead eggs to turn blue.  These two things led me to believe the blue ones are dead.  I haven't removed any eggs yet because I am not sure which ones are good.  Haha!

That problem solved itself, the blue ones indeed were duds. They disintegrated sometime today clouding the water in the tumbler and stinking. So I did a 100% waterchange just now, and accidentally added 2 drops of MB instead of 1. I counted 24 eggs remaining, but they are difficult to count all clumped together.  There were a few eggs that were loners (free floating by themselves) rather than clumped together, and I noticed all the loner eggs died today.  Maybe as singles they get bashed around by the air too much?  If this tumbler doesn't work, my next plan is to try water tumbling rather than air tumbling.

I think 2 drops is better than one.  I have some orchid dottys that are tumbling right now, and I hope for a hatch tonight.

Oh, good luck Kathy!  2 drops it is! 

Not working so well, I think only about 10 eggs intact in there now. I'm thinking the air is too rough for the eggs.

I've never had success with air-driven tumbling, nor have I had success with eggs pulled this early.  I used a fine-tuned water-driven tumbler.  I also see the above setup and wonder - how are you controlling the heat / temp?  

Matt, yeah I didn't think it would work, but I had too much going on and didn't have anything setup to tumble.  I know that tumbling eggs this young is very difficult (has it even been done before?), but I also know that leaving the eggs with him was as good as chucking the eggs in the garbage, so I figured I would try!  :p  I was setting the tumbler in the sump of my broodstock tank to maintain temperature.  I just put it on the shelf to take pictures.  I have 2 weeks now until their next spawn for me to get together a water driven tumbler, and have a kriesel-like design in mind.  In the meantime, I bought 4 single, young adult wild (ugh) Banggai to try to pair myself.
 
I feel that egg tumbling is likely the only commercially viable way of reproducing Banggai, so I'm going to persist.  Handy as it is (not), I have the perfect pair to force me into early egg tumbling adventures. 

I have a very limited experience AI bangaii eggs.  My best attempt was to pull them at day four and I got them to survive until day 17 until I blew them out of the incubator.  I do like your idea of the incubator as that style was my next trial.  The one thing I was going to do was to use the same setup as you except feed the incubator with an aqua lifter instead of air.  I haven't been able to try it out yet as I lost my male bangaii.

Aqua Lifter is a good idea.  I should see if I can find one around here.  I was just going to buy a Mini Jet 202 (or whatever the numbers are).  Nice thing about the Mini Jets is they are easily adjustable.  Sorry to hear about your male...that is so frustrating and saddening.  Unfortunately, my male won't hold longer than about 36 hours with most of the spawn gone by 24 hours so I have a small window to work with.

EasterEggs, would the idea shown in this video work for you?
 

I have not tried it, but I was going to look at doing something similar.
Flow can be adjusted by the pump strength and position.
Various container shapes could be used.
 
The other similar principal I have seen, is using an air driven tumbler, with the airline above the egg compartment.  The egg compartment is screened at both ends and is open at the bottom.  This causes an updraft which the eggs tumble in.
 
Im only just starting to look into this so, its just idea's not actual working items.

ops. double post

Darren, that is very cool!  Thanks for sharing.  I couldn't figure it out at first until I realized the tank is full of water!  The water is so clear it looks like the tank is empty.  I thought at first the glass was overflowing.  I really like how simple this is, and the fact that I have everything needed to make this work.  I think I will ditch my kriesel-like design idea, and try this one next!
 
The second tumbler you describe as air driven without the bubbles touching the eggs is a design I used in the past with some success, but my particular design wasn't really good enough.  Maybe with some tweaking it might work as well.

Mindy, check this out: http://www.mbisite.org/Forums/tm.aspx?&m=31899&mpage=2
I used a 1L flask for dottyback eggs and it worked pretty well.

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

Thanks Tal!  I haven't come across those pictures of your before (or maybe I don't remember since tumblers weren't my focus months ago haha).  That's a nice, simple design too.  I forgot to look for an Aqua Lifter pump while I was at Petsmart.

I got the idea from Chad Vossen who was using a beaker over a sump. I thought a flask might provide a better flow pattern due to the shape. Seemed to work well. Getting the rigid airline placed correctly was a bit of a challenge but well worth it.
 
I used fiberglass window screen to cover the top, that kept the egg mass in the flask but allowed the larvae to get out after hatching. The one mistake I made was not covering the intake on the pump on the first try...

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

Brilliant idea with the screen allowing the larvae out when they hatch!

 
This is basically the same as the water driven version I used, but the flow they're showing is substantially more than anything I did.

Has there been any research or does anyone have any info on why eggs pulled early do not do well typically? If we could drive to the true root cause of that then perhaps a system could be designed around it. I am wondering, is the cumulative effect of all the collisions in a tumbler affecting the eggs? If so I am thinking of trying a system where I have the eggs sandwiched between two kayers of mesh. Then use a wavemaker to alternate current flow both directions past the trapped eggs. Just trying to think of what the males might be doing, or not doing, differently than we are.

Dan, Banggai eggs stick together in a mass, they don't tumble individually like whatever eggs those are in the video.  So Banggai eggs won't be bouncing off eachother.  I'm not sure that anyone knows why Banggai eggs collected so early typically don't survive, otherwise we would already know how to remedy the problem.
 
I'm hoping to figure out what this problem is; this is my goal.  I am thinking that the relatively long incubation period is a major hurdle, but what about the length of time is possibly hindering the eggs?  I'm not sure.  I have a few ideas though.  Maybe it is physical injury to the eggs?  Bacteria?  Prolonged light exposure?  Inadequate oxygenation?  Maybe the male excretes something in his mouth that contributes to incubation or development?

I was wondering about light as well...

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

I didn't realize they stayed attached during the tumbling. I guess I should watch more YouTube videos.

Maybe if we can go back and try to pinpoint what the male does during his holding then it will lead us to better survival % of early eggs as well as those pulled later.

My banggais only spawned once prior to the male killing the female (during a botched spawn I believe) so I don't have a lot of observation time. My thoughts are the male is moving the eggs inside his mouth and pumping water across them in and out. As Mindy and Tal mentioned above, in darkness. How can we simulate all this? If someone could do some microscopic examination of the eggs in like several hour increments maybe we could see "how" they are failing if you will.

I would be quite happy to do an observation as you describe, though I don't have a good enough microscope.

...all part of my proposed research for Banggai Rescue...if the timing works out
 
I should mentoin, I tried to post a second response but the power flickered and killed it.  The african cichlid style tumblers that are air driven with a wider chamber and mesh at top and bottom typically don't work.  Since the eggs are one big mass, they're either not tumbled enough, or they get pulled against the top screen.  This is why I went to a water driven tumbler and ultimately found success.  There's a myriad of ways to accomplish this basic design, although I have to say, seeing it in action, I think a wine glass may be hands down one of the best ways to accomplish it...going to have to look around here more for suitable glassware.

Yeah, I experienced the same thing with the "cichlid style" tumbler...had quite the difficulty getting the eggs to tumble properly, and was more luck if they tumbled ok for long enough for them to hatch (only a few days with that pair).  Looking at the wine glass idea, I'm thinking a "tumbler glass" might make a great tumbler.  Haha!  Essentially a wine glass without the stem so it could be placed into a smaller vessel.

Hmmm....I forgot about that wine glass tumbler idea.  I need to try that.
 
I tried using a PhosBan reactor with my Banggai eggs which worked significantly better than the ghetto pop bottle tumbler.  It takes a lot of flow through the PhosBan reactor to keep an egg ball tumbling though - a MaxiJet 1200 is barely enough.  I collected a spawn about 2 weeks ago, and the eggs lasted 6 days.  So there is some progress.
 
I ordered an Aquabreed 200 and an Aqua Lifter Pump from BRS to make a water driven tumbler.  I have a feeling once I figure out a good method of tumbling the real challenge will be to keep the water quality acceptable for the extended incubation period of the Banggai Cardinal.  I think that will be the tough part.  So far, I'm just using methylene blue.  UV will probably be the next step.

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

The water tumbler I used, and evenutally hatched s group of PJs in, seemed to work well.  I am not sure if I ever posted an update with it in my PJ journal.  Basically an very tall (18" or so) vase that I found at walmart for $8 amd I use a maxijet with a valve to pump the water down.  The top of the vase is considerably wider than the bottom, so by the time it hits mid vase, there is not enough water pressure to push it up any higher.  The eggs bounce between 2/3 and 1/8 hight of the vase, and very gently.
 
just a thought.

That's an idea Eric.
 
I actually split this pair up, put the male in my reef tank, and substituted a young male about a week ago.  Time will tell.  The female should be ready to spawn soon.  They have been very flirty right from day 1 together.
 
I have a second young pair that is only holding just shy of a week so far (holding second spawn right now).  I will give him a few tries, and if he isn't getting better with time I will start using his eggs for AI adventures (lol).

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