The Lightning Project

The ongoing saga of the PNG Lightning Maroon Clownfish Breeding Project

Filials, What the F?

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It seems that at least one irate aquarist thinks I’ve made up the concept of “F1″, “F0″ and so forth. Maybe as a marketing ploy even?! Nope, sorry, they’ve been around forever.

Originally, this content was intended for inclusion in the Conservation Breeding Chapter of my half of the book, Banggai Cardinalfish, which was published last year. Most of that last chapter was cut…we were 17 pages over-length and the most esoteric parts of the book were the first to go. But, I found the text in my drafts, and I’m publishing it here, very slightly edited from what I found in my drafts.

Filials Denote Generation and/or Inbreeding

You will routinely see the use of tags like F1, F2, and F3 attached to aquarium-produced specimens of wild-type forms in the freshwater trade, most often attached to fish from private versus commercial breeders. These notations are called filials, and are meant to convey the generational distance from the last “outcrossing,” that is, a mating between two completely unrelated fish.

Officially, F0, or generation zero, represents a pairing of two unrelated fish regardless of the source, and the resultant offspring are F1, the first generation. Mate those F1 fish together and you have F2, and so forth. However, in some cases, including in the general aquarium hobby, F0 more often used exclusively to mean a wild-sourced broodstock. This slight difference in use can cause a significant level of confusion.

From an inbreeding standpoint, and for tracing the extent of inbreeding, the use of F0 to denote the parents of an unrelated mating is both practical and proper. From a conservation standpoint, breeders are often concerned with “distance from the wild.” Given the possible conservation implications, and that the general accepted practice among freshwater hobbyists is to use filials to show distance from wild genetics, Banggai breeders might consider implementing the methodology outlined here.

F+ the number of generations away from wild-sourced genetics. Thus:

  • F0 denotes a wild fish.
  • F1 is generally the progeny of a wild pairing.
  • F2 is the offspring of an F1 X F1 pair, related or not.

When fish of different generations are mated, the resultant offspring are F+1 to whichever mate is already further away. For example, F3 X F5 = F6, not F4.

Problems arise when a captive line is outcrossed back to a wild fish. This fundamentally resets the inbreeding that has occurred, so F0 x F6 should be called F1. Many aquarists and breeders may disagree, considering such a “reset” as deceptive because only half of the parentage is wild, and thus insist that F0 X F6 should be denoted F7. Some breeders sidestep all this debate, using F0 for wild fish, F1 for progeny of wild fish, and “aquarium strain” to denote everything else.

The important final message is that filials are used by different people in different ways. From a genetics standpoint, the filial is hardly the final word, because it can mean two fundamentally different things: one breeder’s F6 fish may be far more genetically solid than another breeder’s highly inbred F3 specimens, and a third breeder’s F1 fish might be 10 generations removed from the wild. If there is any doubt, a conversation with the breeder should help clarify the meaning of the filial label he or she has applied.

This morning I received an email containing a text conversation from a concerned aquarist, asking me to confirm the claims made by an individual on Facebook who had offered to sell 50 “F1″ Lightning Maroon Clownfish. It is fair to say that the concern was warranted.

The “seller”, operating under a Facebook account with the name “James Cohn”, caught attention with an initial post looking to sell 50 F1 Lightning Maroons, made to a small Facebook group of Clownfish enthusiasts. That post is now gone, removed by a moderator it would seem, but the individual who forwarded the conversation was interested in what was being offered.

I must disclaim the following – virtually everything said here with regards to me is untrue.  I don’t know who James Cohn is, I don’t even know if that is this person’s real name.  For that matter, I cannot even 100% say that I am not being set up here being shown a fictional conversation (granted, that would require a lot of effort to simply create a conversation solely to either discredit this “James Cohn” or to otherwise get my attention). I may well be getting “trolled” here. Maybe “James Cohn” is the victim, being told things that aren’t true by some other 3rd party. This is all, in the end, smoke and mirrors. Or more likely, this is simply someone looking to rip people off. Wouldn’t be the first time.

Regardless, this scenario demonstrates a rising problem I’m seeing in the clownfish world. This is basically Tulip Mania, on a somewhat smaller scale.

I’m watching fish being traded like playing cards, breeders wantonly attempting to hybridize Lightning Maroons with ANYTHING and EVERYTHING, and quite willing to risk the long term genetic integrity of the PNG lineage of these fish by hybridizing them with Gold Stripe Maroons (they almost seem to be doing it specifically to assert their right that they can do ANYTHING THEY PLEASE with complete disregard for the consequences). There are people misrepresenting themselves as legitimate businesses, or claiming to be breeders who may have never reared a single fish in their lives, so as to procure fish from sources like Sustainable Aquatics and Sea & Reef Aquaculture, undercutting their local fish stores to make only $0.10 on the $1.00, flipping fish like cheap Chinese junk at the dollar store.

Venues like Facebook are facilitating this at an alarming rate; when “players” like “James Cohn” (presuming the “h” is silent!) get found out, they can easily reinvent themselves with a new group, a new pseudonym, a new eBay account and keep trucking. Anyone can make themselves look huge, quite quickly, and dare I saw Florida seems to be the largest hotbed for such activity at this time. Facebook seems to overrun with no-name sellers who you’ve never heard of, everyone and anyone being a clownfish breeder who will sell you just about anything you want for the right price. Nevermind the ongoing concept of trumped up, make believe, “artificial rarity”. This latest example, of “James Cohn”, simply being perhaps the most egregious example (if in fact legitimate at all).

Update – while the above commentary is harsh, I am calling it as I see it. I do acknowledge that small businesses have to start somewhere, and more so than ever, it’s in someone’s basement or garage.  I’m a “Basement Breeder” myself after all.

Before I reveal the conversation, let me deal with some of the claims made by “James Cohn”:

Claim #1 - James Cohn claims that “I’ll be getting the pair from the creator of the lightning maroon project”. FACT - there are only two spawning pairs in this fishroom, and neither are for sale at this time. So unless someone is planning to break into our home to steal them, this is a flat out lie.

Claim #2 - The story now changes, and James Cohn claims “Can’t hook you up direct from Matt but I can get you second degree offspring” – FACT – it is possible that other Lightning Maroon pairs are starting to spawn from the first released fish, so this could be true.

Claim #3 – James Cohn then states “Haven’t gotten the pair in but we’ve been in touch with Matt, Zachs a personal friend of mine.” FACT – fact is this – NO ONE has been in touch with me about getting a pair of Lightning Maroons from me because, well, for starters, ALL my offspring get sold through Blue Zoo Aquatics.  There have been exceptions, fish that have been donated to events by me for fund raising, but that’s far different than ME selling fish directly to a private individual.  That DOES NOT HAPPEN, I run my fishroom strictly wholesale to the trade and fellow breeders who have demonstrated the same committment to the trade. In short, there is no way a private individual who’s claimed to live in PA is purchasing ANY fish directly from me.  Never mind a proven spawning pair of Lightning Maroon Clownfish.

Claim #4 – when asked “So you are working on the project with matt”, James Cohn replies “Naw but he’s personally selling me the pair. I’m a 3rd party breeder with ties.”. FACT - I’m not personally selling a pair to ANYONE, but particularly not to a “James Cohn” in PA. The only way ANYONE is able to obtain a pair of my fish at this time would be one of the aforementioned avenues – Blue Zoo Aquatics, or the off chance I donate a fish to an event.

And it goes on from there.  Nevermind that James Cohn is using MY photography to peddle his nonexistent fish onto unsuspecting aquarists on Facebook.

In short, AVOID AT ALL COSTS. If anyone claims to be selling fish “directly from Matt Pedersen” by all means, ASK ME FIRST.

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In the very week morning hours (eg. 2 AM) on 12-13-2014, I discovered spawn #47 from the wild caught pair.  Obviously it was actually put down on Friday afternoon (12-12-14).  Now that the fish are off the Chloroquin-laced food, it’ll be interesting to see if this clutch holds out or not.

As promised, I finally got time to look at the offspring that Mike Doty had managed to rear from my F1 Lightning X Lighting Maroon Clownfish pair.  While Mike had initially stated he felt the split was 50/50, today’s visit showed something different.

All told, I only managed to count 3 white stripe offspring in the BRT. Meanwhile, there appeared to be 6 distinctive Lightning Maroon-type offspring. What I didn’t see were any fish that, at this point, looked atypical from either known phenotype.

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With only 9 (approximate) juveniles, the sample is far from conclusive. The 3/6 split could be representative of anything from a 50/50 split to a 25/75 split. A new phenotype could be missing because none simply survived in this first successful run.

That said, the 3/6 split, if a valid sample, would represent something we perhaps don’t want to  see.  It would imply that Lightning is straight dominant, but it would also imply that a double-dose of Lightning is in fact fatal to the offspring and they fail to develop.  This is one of the current working hypotheses for the Snowflake gene in Ocellaris, and anecdotal reports continue to bolster that line of thinking (reports are anecdotal in so much as a breeder saying “It’s about a 60-70% snowflake result” is not the same as a breeder saying “I got 140 Snowflakes and 65 wild types in this clutch”).

It is fair to say that there is still hope for these fish – they are QUITE YOUNG and all we can truly discern at this point is that they outwardly either are white stripes or aberrant.  My tune could change as these babies develop, or if subsequent results are different. It is not surprising to me though, to see similarities emerge in various mutation types.  We already have Picasso/Platinum and the sister mutation of DaVinci/Wyoming White. Much as I wonder if these twin mutations could represent the same gene in different species (or simply a similar genetic mutation in sibling species), would we come to find out that Lightning is in fact not unlike Snowflake? After all, genetic analysis has revealed that the Maroon Clownfish are in fact very closely related to the Percula/Ocellaris complex, so it would not be surprising to find similar or the same genes present in all these species based on their common shared ancestor.

It is also an interesting footnote to observe that these tiny juvenile clownfishes DO represent a the first F2 generation of Lightning Maroons.

Here’s a quick rundown.

The 5th spawn of the F1 Lightning X Lightning Maroon Clownfish pairing was collected and hatched by Mike Doty while I was away.  In short, he scraped off the eggs, hatched them in a 1 gallon jar in a water bath with simple aeration, 75% clean new water.  Come November 28th, Mike relayed that settlement had started.  The moment we’ve been waiting for was here – is there something new?

Well, Mike’s first words were “About 90% sure we have some normal striped fish.”

This, of course, does the following:

  • likely rules out Lightning as a simple recessive gene. If it WAS recessive, then both parents would be “double dose” aka. homozygous, represented as l/l, which means that each parent could only contribute a recessive lightning gene, and thus, each offspring would also get one copy each, one from mom, one from dad, and thus, could only be l/l as well. For the moment, while another couple test matings will bolster the data, the fact that there are white-stripe offspring pretty much precludes this being a standard single allele, single locus, recessive trait.
  • does not rule out straight dominance. If it was straight dominance, and each parent is “single dose” aka. heterozygous, represented as L/+, then 25% of the offspring would not get a gene from either parent, and thus, 25% would be white stripe maroon clownfish.
  • nor does it rule out partial dominance. This of course, would work the same way as dominance, except that 25% of the offspring would get a lightning gene from EACH parent, and would be homozygous for Lightning, represented as L/L. This is the scenario that most people are hoping for, because with the new homozygous state, there comes the potential for a new phenotype that could be different from the Lightning that we know.

As of today, 12-8-2014, I spoke with Mike briefly and have to relay this news – while he doesn’t have many babies left, he believes that the phenotype split is roughly 50/50.  That is to say, half white stripes, half lightnings.  So far, he also has not seen anything unique or new in this F2 generation.  I have yet to see the babies for myself, and have yet to take pictures or do a headcount, but these cursory, informal results, mirror another clownfish mutation that seems to not fit the mold as we’d expect – SNOWFLAKE in Ocellaris.  It’s my hope to get over to Mike’s today yet to see for myself.

In other news, the 6th spawn of the F1 Lightning X Lightning pair was put down on 11/29/2014. It appears I finally won the battle of the tiles:

LxL Spawn #6

LxL Spawn #6

Meanwhile, I brought some new clownfish into the fishroom earlier in November (the 19th and 22nd) and was trying out the new Ick-Shield food from New Life Spectrum.  This is basically a Chloroquin-laced pellet food that is meant primarily to prevent disease such as Crytopcaryon, Amyloodium, Brooklynella etc…pretty much the things which are sensitive to the active ingredient. I decided to not simply feed this fish to the new arrivals, but also to feed it to one of my holding systems AND the wild Lightning Maroon and her mate as preventative medication, just in case.

Well, it turns out that there is an unfortunate side effect to this feed; it seems to shut down breeding activity.  All my routine pairs stopped spawning. The Lightning and her mate did finally put down a spawn on December 1st, 2014, #46.

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Unfortunately, it appears as though the spawn was not fertilized…the eggs didn’t develop, and after 48 hours they were gone. A few days after that, I read, anecdotally, that Chloroquin can cause male sterility??? Not permanent according to the rumor, but certainly a potential setback. As far as the efficacy of the food, my jury is out. The larger fish which were feeding well on it by and large remained disease free, but not all did.  I still had a Brooklynella outbreak, although not in the fish I would have necessarily expected. Once that outbreak started, it then affected other fish as well despite their feeding on the pellets.  I’m also seeing either Cryptocaryon or Amyloodium on fish which were visually “clean” upon arrival, which were in dedicated QT systems, feeding on this food from day one.  So the question here is were they simply getting ENOUGH feed as they were small fish which cannot readily eat the small pellet size.

So of course, one is left with questions, not answers.  There is no way to say the food didn’t work, nor is there any way to prove that it does work. Absence of disease is not proof of prevention, that much I know for certain. Lack of a cure, or lack of prevention, which IS documented, only raises questions about why it didn’t work as suggested and certainly requires investigation (eg. would a smaller pellet size be better accepted…could these failures stem from simply lack of feeding, or lack of sufficient feeding, thus insufficient dose to the fish?).

Circling back to LxL Spawn #6, as the week progressed an interesting change in behavior occurred starting around December 4th, 5 days post spawn.  The larger female F1 Lightning became belligerent towards the male, and over the day drove him from nest tending duties.  December 5th, a Friday, would have been 6 days post spawn, and the night of the first hatching.  I was simply swamped with preparations for sending our dog to live with my brother, and failed to pull the tile.  By morning, Saturday, December 6th, 2014, it appeared that I had not missed much…most if not all the eggs were still there. The pair remained at odds.  We left for the weekend to ship our family dog, and upon Sunday, December 7th, there were still a few dozen eggs remaining, although they appeared potentially dead and disappeared throughout the day.  By nightfall, the pair was starting to be less antagonistic, but I am still keeping a close eye on them.  Hopefully, we’ll get another spawn soon – this was the only mature pair in the house that didn’t receive Chloroquin-laced foods (as they don’t reside in the fishroom with the rest of the fish).

Just jotting down notes…Spawn #45 of the wild Lightning Maroon pair happened on the evening of 11-19-2014.  Meanwhile, LxL (the F1 Lightning Pair) Spawn #5 was gone by the same day….they obviously didn’t like me tiling their entire tank. I need to go by more terra cotta flowerpots…maybe they’ll like that.

So much going on and I’m NOT doing a proper job of tracking things.

I should start with the unrelated F0 PNG White Stripe Maroon Clownfish pair…they spawned sometime last week, possibly Wednesday the 12th. I didn’t make much of a fuss about documentation of it because I knew I couldn’t deal with them at any rate; I can’t remember if it was very late Sunday night, or more likely Monday afternoon, when I found the last few eggs just sitting around in the tank.  This pair, overall, is spawning quite infrequently.

The Lightning Maroon Spawn, #44, never made it to hatching.  Something happened, and the pair consumed the eggs.  Could the Lightning foundation pair be winding down or did they just have a bad spawn?  Their prior spawn produced a good amount of juveniles, which were undergoing settlement and on TDO A by last Thursday, which is when I drained them down to about a gallon, filled with clean new aged saltwater, and then turned them on to the larviculture system.

LxL spawn #4 was one I was going to let go because I wasn’t in town (I was off speaking at the Milwaukee Aquarium Society’s “Fish Bowl” event), but in short, Mike Doty decided to play with harvesting the eggs.  He took approximately 25% of the next out, using my little egg siphon, on Friday night (Nov. 14th).  I got a text while in Milwaukee…there was a hatch..and a big one.  Surprisingly, he simply did it in a 1 gallon olive jar with an air feed, sitting in a water bath with probably 75% new water.  If I had done that, every last egg would have been dead.  All credit where it’s due – Mike may well in fact rear Lightning X Lightning offspring before I do (they’re at HIS fishroom ;) )  Mike further relayed that this was a split hatch, with more eggs present Saturday the 15th, but all gone by 4 AM Sunday morning.

Yesterday, I caught wind that the F1 Lightning X Lightning pair was going to spawn again…the female was roving the tank with her ovipositor down.  I did my best to FURTHER add tile to their tank, blocking their prior spawning spot off..which caused them to move to another side of the tank’s back wall, which I then blocked off with tile. They were still going through the motions of finding a spawning site when I turned the lights off after Midnight; technically they spawned in the overnight on the wee-early morning of 11-18-2014, but if I was to be thinking about these eggs properly, then normally they’d be a 11-17 spawn.  Ultimately, they selected the ONLY section of black plastic background that I cannot get tile on (near the tank’s filter return).  Either the nest was small, or they ate most of it, and I am considering foregoing working with it.  I’m thinking that what this pair may really need is a clay pot…that might be the ticket here.  So LxL Spawn #5 has been put down, but for now, probably will be left alone.

So despite two nights of attempted hatching of harvested eggs, I ultimately only got ONE viable larvae out of LxL Spawn #3.  It is in a BRT all by itself.

On 11/7/2014 – the F1 Lightning X Lightning pair threw down another nest…and once again, not on a tile.  I won’t be in town to try trapping or harvesting this nest, so I’ll have to wait for the next spawn. Will update with some photos as I’m able.

Not a lot of time tonight, so I get to be real quick.

Spawn #44 was laid by the F0 pair on late evening of 11/3/2014.  Spawn #43 ended up no hatching much when it was finally set up, but seems to have hatched apparently during the morning.

I opted to harvest 50% of the nest from the F1 Lightning X Lightning pair tonight, the 7th night post hatch, the overnight from 11/3 into 11/4. I used a very simple harvesting technique, using a section of air tubing, inserting the tip section from a 2 ml pipette, which was cut at an angle on the harvesting end.  The eggs were scraped from the back wall while the tubing was siphoning into a specimen cup.  The harvested eggs were added to a brine shrimp hatchery resting in a water bath in a BRT, with a bubble rate of a few bubbles per second.  3 larvae were dislodged from their eggs during harvest…one was viable and started swimming, while the other two appeared lifeless.

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.

 

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