The Lightning Project

The ongoing saga of the PNG Lightning Maroon Clownfish Breeding Project

Browsing Posts tagged larvae

It’s the 7th night for Spawn #43 which was spawned on 10-22; we’re on the night of 10-29 going into 10-30 now; there wasn’t any noticeable hatch on the tile over the 6th night (I left it with the parents yesterady)….it will get pulled.  This is what Spawn #43 looked like when it was laid fresh on 10-22…I should also point out this is the first time I’ve taken pictures of the F0 original wild pair of Lightning Maroon X White Stripe PNG Maroon Clownfish.

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The larvae from spawn #42 that are still alive have largely settled out…it was a small group, but I’ll find a way to work with them.  More importantly, it looks like LM X LM Spawn #2 might be over…I have not found any larvae in the BRT, so one more look, I repurposed  it for Spawn #43.  The eggs got the usual dip in H2O2, but started hatching during it, so they were moved straight into the BRT with 8 gallons of broodstock water and a few ML of RotiGrow Plus (only thing I had thawed…it’ll be fine for the first 24 hours and by that point I’ll have RotiGreen Omega up and running).

The most frustrating news is arguably LM X LM Spawn #3.  So..to recap.  Here was the first spawn:

F1 (Lightning X Lightning) first spawn for the Lightning Maroon Clownfish

 

So…I took some proactive steps and tiled the back wall of the tank.

This was LM X LM (or LxL if you prefer) Spawn #2 on the first attempted hatch night:

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Yeah..that gray patch on the side..those are the eggs…

So that didn’t turn out obviously…so I tiled the side wall as well.

Here is Spawn #3, laid on 10/27/2014:

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The internet shorthand FML seems somehow appropriate. Clearly the pair is flipping me the middle fin.

 

So Mike Doty has had comparable luck to some of my better “early runs”; Spawn #27 remains at his house and in a startling role reversal, I’m now watching HIS fishroom.  As of yesterday, I discovered the first post-meta offspring, a Lightning juvenile, in the warmer, more lightly-stocked right BRT.  The rest are still pre-meta fish.

Spawn #28 was pulled too early, and I should have listened more to Mike Hoang’s advice to do an H2O2 dip on eggs that don’t hatch in the first night.  It seems that the eggs remained viable on the tile up until yesterday afternoon, 4-8, but the majority never hatched. Interestingly, it wasn’t until the morning of 4-8 that the last of the eggs in with the parents had disappeared. There are a scant few larvae in the 10 gallon tank from this batch as a result.

The other news to report is that on the afternoon of 4-8-2014, the 29th spawn was put down by the Lightning Maroons.  Based on my dates, I should pull the tile for hatching next Monday, the 14th.

Talking with Mike on Friday, as best as we can figure out Spawn #25 was put down Saturday, February 22nd. This fits the timeline…come Friday night (Feb 28th) we’d be at 6 days post spawn, which is the earliest eggs hatch. I opted to wait until the 7th night, which was last night (Saturday, March 1st) to pull the tile.

Come Saturday morning, it was clear that some eggs were gone…perhaps as much as 50% of the nest must have hatched on the 6th night. But as we’ve seen so many times before, these eggs, for whatever reason, are prone to split hatching over 2 nights (a problem I also had and never fixed with my wild Onyx Perculas). So last night, technically 2AM this morning, I pulled the tile to prepare it for hatching.

I returned to the 15 minute hydrogen peroxide (H2O2) bath in broodstock water, and I drained the 10 gallon clownfish hatching tank, soaked it with FW, rinsed it all out, and filled it with 5 gallons of broodstock water. Thinking that either the use of clean new water, or a weak air setting, caused the hatch failure in Spawn #24, I set up the tile with a rather vigorous flow of air from the wooden airstone. A dime white moonlight at the far end of the tank was set up to draw larvae away from the heavier currents & flow of the hatching setup and tile.

By 3 AM, I took a peak and noticed there seemed to be a good hatch, so I harvested 1.5 gallons worth of rotifers and added 20 drops of RotiGreen Omega. I was feeling optimistic.

9 AM this morning, I checked in and removed both the hatching tile and the large tile that I had placed on the bottom to prevent the hatching tile from sliding / fall. To my disappointment, it appeared that most all larvae were on the bottom, and some were dead. There was only maybe a dozen eggs on the tile that had not hatched. I added almost 50 drops of RotiGreen Omega to help green up the water and hopefully get the babies up off the bottom; I also turned the air flow down significantly to just a trickle, feeling that perhaps I should have done this “last night”. Could it simply be that several hours of turbulence in the tank had caused some of the larvae to expend excessive amounts of energy and doomed them to a quick death?

I’ll be keeping an eye on these obviously….wish me luck.

Tonight, after several nights of battling my larval rack, I finally pulled of the split of Spawn #20 into two separate groups. Long story short, I had taken off two BRTs off a shelf that had somehow “Dropped” on the one side. Both BRTs got a full scrubbing followed by a wipedown with hydrogen peroxide. Meanwhile, I spent 2 nights and 3 days banging on the shelf with a rubber mallet until I finally freed up the shelf from the plastic locking mechanisms. Thankfully I saved some of the extra plastic parts that came with the epoxy-coated wire shelves, and was able to reset the shelf height and set the BRTs back up.

Once the larviculture tanks were set back up, it was easy to simply scoop out half of the water and larvae from the 10 gallon tank. I took a photo of the larvae in the bucket so I can count how many I moved into the BRT. Just as I scooped the babies from the 10 gallon into a 5 gallon bucket, I in turn did the same thing to introduce them to an empty, sanatized BRT.

I’ve continued with doing daily water changes on the larvae and have, at this point, dropped the specific gravity from 1.025 down to 1.018. I’m also doing nightly infusions of rotifers that have been enriched for an hour or two with Super Selcon.

In other news, Spawn #21 was put down this evening (1-9-2014) by the Lighting Maroon pair.

The hatch of Spawn #20

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Well honestly, I just don’t know what I did that caused this one to be so much better than any prior to date, but somehow the stars aligned and I wound up with something like 1000 baby Lightning Maroons.  And the best part is that I am HERE and able to care for the fish well through metamorphosis.  God help me if I screw this one up (which is a real possibility given so many babies in a 10 gallon tank…I *may* wind up splitting the batch).

As you may recall, I pulled the next on the night of 1/1/2014 going into 1/2/2014.  I re-read all of Wilkerson’s text just to see if I had missed ANYTHING that could be helpful, and one line may have been the difference.

“If in doubt, err on the side of too much aeration” – Joyce Wilkerson, Page 190, Clownfishes, third printing (2003)

Well, I took that to heart and didn’t dial back my slintered glass airstone…I just let the thing rip faster than I felt comfortable.  And that might have been the ticket…it “feels” like that’s the only variable I really changed.  That said, I didn’t sanitize the eggs this time, and I started the tank with a mixture of new water and broodstock water vs. all straight one or the other.

In any case, I got the kind of hatch I get most every time with other clownfish species, so maybe I really just need to be more aggressive with my aeration?  I’ll obviously follow the same pattern on the next batch and see if it repeats.  A whole 2 years almost to dial in on the best hatching protocol…damn…you’d think I was a newb at this.  Who knows…

The Hatch of Spawn #20

The Hatch of Spawn #20

This will be very short – I’ve just been swamped with freelance projects and endless deadlines and well, there’s just nothing good to report on the latest runs.

Spawn #12 did very well right up until metamorphosis.  Immediately after metamorphosis, every larvae would die.  I don’t know if it’s the different brine shrimp eggs I’m using (or even the fact that I’m using brine shrimp vs. not). It could also have been that I didn’t rear these with 24/7 lighting, but instead let them run on the regular photoperiod for the rack of tanks they were in.  Water quality remained spot on and the fish were well taken care of.  Thus ends Spawn #12.  Probably changed too many variables all at once, but at least I may be onto something regarding getting good hatches for these eggs.

Spawn #13 never did produce much larvae, wound up hatching out over 3 nights.  I found myself somewhat depleted on rotifers with this batch coming so quickly on the heels of spawn #12, so this batch really never stood a chance.  I augmented from day one with Otohime A, introduced brine shrimp as early as was reasonable, but found no larvae made it to settlement.

Unfortunately I may have put the pair off spawning for a little bit (might not be a bad thing while I regroup).  The lights on their system had been going off incredibly late in the night (eg. 3 AM) so I shifted the photoperiod. No spawns have come following this, which isn’t really a surprise.  I’ll be curious to see how far back this pushes them.

Meanwhile, as I just posted on Facebook, I have more Fire Clowns (Amphiprion ephippium) than I know what to do with.  Figures…

8 days post hatch.  To brine, or not to brine, that is the question.

First noticed this at roughly 1:30 PM on July 7th, 2012.  Dare you to hold your breath…

Larval offspring from the Lightning Maroon Clownfish nearing settlement.

Lightning Maroon Clownfish larvae, nearing metamorphosis

Headstripes forming on larval Maroon Clownfish spawned and hatched by the Lightning Maroon Clownfish

So this amounts to a chronological retelling of the story to date, this time with photos, starting  a couple weeks back now.  Perhaps not in as much detail as my minute-by-minute updates, but a good overview of the run to date.

June 21st, 2012

The ongoing health problems with the Lightning Maroon remained, and the left eye on the Lightning Maroon was showing slight swelling.

On a day initially planned to do a skin-scrape of the fish for further examination, I had to call things off because the fish had started going through pre-spawn motions.

By the time we had finished doing a skin scrape on some Banggai Cardinalfish downstairs, Barb & Heidi from the Great Lakes Aquarium got a super special treat, seeing the actual nest having been spawned while they were here.

Lightning Maroon Clownfish Spawning & Eggs

June 22nd, 2012

I was genuinely worried whether we’d have eggs 24 hours in.  Thankfully, they proved to be good parents and good “clownfish”; the first spawn egg eating proved to be the typical first test run that so many clownfish seem to do.  This batch, while I didn’t get a good photo of the parents, was doing well.  The swelling on the Lightning Maroon’s eye had gone away.  Phew.

June 23rd, 2012

So much for resting easy about the health of the Lightning Maroon. The eggs were developing (a fair number that probably were infertile or diseased were removed by the pair), but some funky gunk (yes, that’s the scientific term) showed up on the Lightning Maroon’s right face.  I was once again on high alert; this wasn’t pop-eye; this was more reminiscent of the mouth-rot I had to battle back a little while ago.

June 24th, 2012

So much for being on alert.  By evening, things looked so bad on the Lightning Maroon’s face that I pulled the trigger and initiated the third course of treatment with Maracyn SW and Maracyn II SW in this system.  The telltale bulge around the right eye had started to show as well.  I felt I had little other option at this point; this fish is simply too valuable to take a wait and see approach when symptoms like these show up:

The eggs were looking good and developing fast, although I took little comfort in that given the current situation with the Lightning Maroon.  The roller coaster of stress over this fish during the past couple months has been excruciating.  No doubt, there were times I pondered whether it would all be easier if the fish just passed away – of course solely a passing fancy, but when things are clearly out of your real control, it is incredibly tough to sit there and do “what you can”.  Of course, it’s a whole new level now that we are well within sight of the next major milestone in this 2+ year long project.

June 27th, 2012

June 27th represented the 4th day of Maracyn + Maracyn II treatments, and once again, it appeared I had potentially averted a crisis or loss.  The condition of the Lightning Maroon was drastically improved.  The eggs…the eggs were showing eyes?  They had the classic silvery look of clownfish eggs before they’re going to hatch.

I had been worried that these eggs would be hatching out while I was on a trip to Boston to speak at the Boston Reef Society; but now, only 6 days post spawn, I was very worried that a hatch could come sooner than expected.  The signs (and the data out there) said it was possible, sure, but maybe not likely?  Still, if I waited too long and did nothing I could miss the hatch. Conversely, if I pulled the nest too early, I could miss killing the eggs before they actually had fully developed.  Honestly though, I felt far less pressure about the decisions I was about to make than any of the disease-related issues with the Lightning Maroon; this is clownfish breeding, I can handle it.

There was really only one route to go – I had to sit and watch the tank.  The lights go off at 12:15 AM, so I got things situated for a possible hatch.  I used a small LED flashlight at the far corner of the tank as a larval attractant.

While waiting for the lights to go out, I prepared the area  with buckets and siphons to take out larvae should they hatch in the tank.

Downstairs, I prepared a black round tub to receive broodstock water and possible babies.

Lights went out, and it was time to wait.  All pumps were turned off through an extended feed timer on my Apex Lite (which would ensure they’d all come back on in the event that I somehow forgot about them and went to be).  I did have to unplug the battery backup on the Vortech…can’t have babies going through that pump either.  I’d check every once in a while, and initially got excited around 12:20 AM when I saw movement in the beam of the flashlight – until I realized it was copepods swimming around.

Many more checks turned up nothing, and I was starting to wonder if I had jumped the gun.  Multiple plans of “what next” rolled around in my head, but they all disappeared at 1:23 AM on June 28th, 2012.

That is not a copepod.  If you can’t really see it, maybe this one will help:

The moment that first baby clownfish showed up, I pulled the tile under almost complete darkness, moving it downstairs in a bucket with a lid and 5 gallons of water from the broodstock tank.  I set it up for artificial hatching, and assumed that come morning, I’d see hundreds of clownfish swimming around.  That was the hope…

June 28th, 2012

So much for hatching overnight.  There was ONE baby in the tub.  Terrific (<-sarcasm).  1 is better than none, so in the interest of keeping the one alive, I was forced to tinge the water green with a very light treatment of RotiGreen Nano, and a very small addition of rotifers (lest the baby starve).

The worst fear is that I had somehow killed the eggs in the move or prevented the hatch, which would have generally killed the eggs overnight.  There was only one way to find out.  I took a quick look at the tile.

And here’s what I saw…

They look perfectly fine.  And what a great opportunity, thanks to the advent of digital photography and Photoshop, to get a headcount.

That’s roughly 310 eggs (each color group represents me counting to 50, with the scattered red dots representing the last 10 I counted).  It’s not an exact headcount, but gives a great approximate number of eggs.  Hardly the spawn of several thousand that some Maroon Clownfish are known to put down, but I’ll take it all the same.  So very carefully, this tile went back into the black round tub…

…So long as the eggs didn’t die, there was still hope.  The rest of the tension filled day was spent fighting the urge to recheck the tile for dead eggs.  Come nightfall, I stuck with the photoperiod that the eggs had been used to, and turned the lights out in the basement a little early so that things were basically pitch black by 12:15 AM on June 29th.  Just after 1:00 AM, a quick check with the flashlight caused me to announce to the world, “Ladies and Gentleman; we’re rearing Lightning Maroon Larvae.

June 29th, 2012.

With only hours before my departure to Boston, I had to get things set up right.  As the night progessed into the wee hours of morning (that we normally still call “night”), I fired up the lights, and checked the tile:

No stragglers – that means a 100% hatch.  That means 300-ish baby maroon clownfish.  300 chances to see something really fantastic down the line.  So long as we don’t botch rearing them!

Mike Doty, a fellow aquarist who happens to live 4 blocks away from me, had been over late (or early if you want to get technical) to see how things were set up and to know where everthing was…well that and to share a beer, toasting this milestone. Mike would be completely in charge of rearing the larvae in my absence.

 

While I got my share of incredulous inquiries about that, I actually had more confidence in Mike than myself; Mike had taken a pair of extra Maroons from me, spawned and reared a couple batches, so he was perfectly qualified in my book (I’ve done clowns, but never maroons before).  We got the larval tub set up with greenwater and rotifers, and in the early afternoon I embarked on my all-day trek to Boston.

 

 July 1st, 2012

I returned home from Boston in the afternoon, anxious to see how things had gone.  Mike had kept me updated via texts during my absence and things sounded good.  The main message I got from Mike was that my three rotifer cultures had failed to keep up with demand, and he had actually depleted his as well.  I wondered, would we wind up losing this batch to starvation?!

July 2nd, 2012

I’m indeed burning through rotifers, but the cultures seemed to rebound and were producing enough for the moment.  The rotifers in the BRT were also clearing out phytoplankton pretty frequently.

Mike and I had set up a drip for the tub using a spare brine shrimp hatchery and a micro ball valve from Julian Sprung’s Two Little Fishies.  Not only is the drip good for top off, but also for introducing foods (phytoplankton) and ammonia control (CloramX) slowly.

Seeing that there were still many babies (some losses, but still many viable larvae), I took a photo for you all; your first look at what *Could be* a larval Lighting Maroon Clownfish, roughly 4 days old.

 July 5th, 2012

Things have gone well, as I’ve slowly doubled the larval rearing volume to 10 gallons, keeping a watchful eye on the ammonia alert badge as I continue to feed 4-5 gallons worth of rotifers into the tub per day.  With the warm basement temperatures (normally in the upper 60′s to lower 70′s, but lately 78F), the rotifer cultures are now roaring; I’m forced to feed them twice daily at a rate of 30 drops of RotiGrow Plus (and 30 drops CloramX).

I’ve done a couple pre-feeding rotifer enrichments with Super Selcon as well, just to keep the DHA levels up. However, today, now just before 7 days old, we reach another step in the rearing process.  Today it was decided the larvae were finally ready to feed on APBreed’s TDO, size A.  And after the second feeding, it was fair to say they are indeed consuming it.

So now we sit and wait.  Any day now, we will catch the first glimpses of stripes as these larval Maroon Clownfish go through metamorphosis and settle out into juveniles.  Most likely, I suspect that even if we have fish that will one day show the “Lightning” phenotype, we won’t see it at this stage in their development.  But at this time, it is anyone’s guess.  If you’re a betting man or woman, it’s time to place your wagers.  Our first glimpse at the possibilities are just around the corner.

I am still owing the world a full post, but in the interest if simply keeping you all relatively updated, here’s some short tidbits.

Based on an egg photo count, there was 310 eggs on the tile and all hatched.  There have been some larval losses, but that is to be expected.  Mike Doty did an excellent job watching the babies in my absence – plenty of live larvae when I returned from Boston this past Sunday, July 1st.

Since returning, larval maintenance has been a study routine of upping the water volume with drips of pre-mixed saltwater (for those who will ask, I am currently using AquaCraft’s Marine Environment – they donated a palette of it for Banggai-Rescue).  The SG is probably running around 1.021.  I am using Reed Maricultures RotiGrow Plus to culture my rotifers, and using their RotiGreen Nanno for greenwater (I may have preferred the Omega variant, but Nanno is what I had on hand).  I’ve been dosing RotiGreen and CloramX (a solution mixed from the powder) at roughly a 2:1 ratio, and averaging 30 drops twice a day now on the BRT. I have been harvesting up to 4 gallons of rotifer cultures daily (2 in the AM, 2 in the PM) to keep rotifer levels up.  As of tonight, we are at basically 6 full days post hatch, so I introduced the larvae to their first taste of APBreed TDO (Top Dressed Otohime), Size A (smaller than the A1 I’m more normally accustomed to using).  All is going well with these larvae, and I look forward to settlement soon.

The Lightning Maroon herself continues to be a problem…the Foureye has been removed for a while now, the Maracyn + Maracyn II treatment was long since done yet low level bacterial problems persist, most recently some very light markings on the male’s face, and then I found what looked like an enlarged light area on the leading spines of the left pelvic fin.  These fish just can’t get a break.  I am continuing to work with Dr. Kizer on some alternate ideas, as we’re really ruling out all the normal causes at this point.  Me, I’m stumped.  Without diagonistics, I think it’s fair to say that Dr. Kizer can’t really offer any other insights either.  We may try yet another antibiotic course, another one dosed through the food, that seems to be where we’re heading.  But I’m also thinking I don’t want to overreact either, so most likely we will try to have the prescription-based feed on hand, ready, should another large-scale problem crop up.

Shot in the very wee hours of the morning on 6/29/2012:

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