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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Wednesday, April 17, 2013 12:01 PM
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I like that idea a lot. We tossed around the idea of holding them down with mesh yesterday also, but didn't have the tubing idea you did. Essentially, make a micron mesh sieve like the one I showed a pic of, with the bottom just being the bottle. And then filling it over the mesh/bottle top and into the tubing. That could work really well! I think my properly sized micron mesh (180uM, IIRC) will be here today-ish so I could do a couple test bottles. The mesh sheet is 12inx12in so I could potentially do many bottles if I needed to. And you are right, a waterchange could be a snap with that little device/system. I would like to use tubing with a bit of substance, similar to the kind I made part of the sieve out of, just because then I could put my palm or parafilm over the top and invert the entire bottle/mesh/tube contraption to mix things up a bit once or twice a day. Floppy tubing could still work if I pinched it while doing that, but not sure if it would work as well (maybe it would be better...who knows right now). The only little snag may be that the bottles have screw tops, so there are those spiral ridges on the outside of the bottle neck. So tubing over top, even if it is very tight, may allow a sort of river to spiral down the outside of the bottle. Maybe it won't, though. I will see what I can whip up (with what I have handy first, then with what I can find at the hardware store for cheap) and give a bottle a test run.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Sunday, April 21, 2013 11:01 AM
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The oil was a FAIL. Most of those bottles were a loss before the oil was added, so most had only a few veligers and were just dumped. Some had enough veligers to bother taking the oil later off, washing them through the 180uM sieve, and washing them into a new bottles. Several bottles from different dates were combined this way. Annoyingly, I was in a hurry so I didn't label that bottle correctly and now it's mixed in with the other containers. This isn't a huge deal since I can just assume they are all the "youngest" ones in most of the bottles. There are three bottles that I know for sure aren't the combined ones. I took some pics of veligers that are 13-14 days old. These were taken from the surface as some had become trapped in a bubble under the parafilm. They weren't all dead yet. I like the first picture because the two veligers stuck together look like the letter "S". It was like a shout out to me A cluster of them: Close up: On the top left of this one you can see the velum Shell and, at the top, the velum again: Velum spread out, in a "bottom up" view. Or perhaps one is the velum and the other is the foot? In veliger diagrams the foot also has little hairs, so it could be I like this pic because you can see the shell opening on the left, the operculum on the far right (just above the green/brown blob which I am guessing is the stomach) and the foot just to the left of the operculum. By the shell opening is the velum. I don't know my veliger anatomy so this may not actually be correct. I've been trying to learn though. It is a bit difficult since drawings don't always translate very well to what you see or think you see under the microscope In a week I will start the settlement experiments. Paige (1986) states there is no anatomy change that indicates competency to settle (as opposed to Aplysia brasiliana that develop pigmented lateral spots and a mantle fold). This week I will start to figure out how I want to approach testing settlement substrates since larval entrapment in the surface will still be an issue, but I need a larger surface area so I can actually access the dish. Cetyl alcohol will come in to play, I am sure.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, April 22, 2013 11:04 PM
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Large die off today in the awesome bottle with the most veligers. I assume it was water quality related. I was concerned about handing the veligers and delayed a water change...of course, bad plan! I changed the water tonight and put some Iso in (I added a bit of tetraselmis last time). Hopefully the remaining will make it the remainder of the week. I combined the majority of the remaining bottles as they all have few veligers. This bottle now has veligers ranging from 4-5 hatch to 4-12 hatch. A bottle from 4-10 hatch had a nice amount of veligers still and I did a water change on that bottle also. Lastly, a 4-8 bottle had a few but I ran out of sterilized water so I will do a water chane on them in the morning and keep them in their own bottle since I want to keep that hatch separate as I could begin settlement experiments with them this weekend assuming they make it. All bottle got Isochrysis tonight. I decided to stick with that for the remainder since those bottles seem to have less air bubble accumulation compared to nanno and especially tetraselmis. I still have not found tubing that will fit over the bottle top yet as Easter Eggs suggested and the time I went to the hardware store I forgot to bring a bottle :/ The cetyl alcohol arrives tomorrow and it can't be soon enough! I think the only way parafilm is feasible is it you have an army of interns and grad students to change it out every two hours 24/7. Perhaps if I was more exact with the amount of phyto added and the stocking density the air accumulation wouldn't be as much, but 1 veliger per 10mL just wasn't feasible for me given the space for culturing. The 4-8 hatch is supposedly competent to settle on the 27th according to Paige. I am not so confident after the water changes today. I put the 180uM mesh in my little sieve for the water change. The majority of them went right through even though they should be plenty big enough to be caught. So either the mesh is not the size I thought, the sieve allows most through somehow along the edges (doubtful since I didn't have this problem when I switched to the 40uM) or they are not growing at the rate Paige states (10uM per day). Perhaps there is a difference in growth rates and competency of the different subspecies? I am not sure but now wonder if they will actually be ready to settle at the end of the week.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Thursday, April 25, 2013 8:27 AM
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Last night I got home to find that the second most awesome bottle (4-10) also had the majority of the veligers drop to the bottom. During the day, I began to wonder if this relates to competency to settle...are they exhibiting the "crawling" behavior Paige mentions? I sucked a few off the bottom with a pipet and put them under the microscope (sorry, no pics right now because I accidentally deleted the program for the microscope off my computer last night and need to reload it) and although some are definitely dead, probably 75% are still alive, just on the bottom. I didn't notice any behavior I would call crawling though, so maybe they are just dying. I have kept inverting the bottle regardless, just in case. I also set up a few settlement trays with small amounts of Spirulina. I took a 100mL petri dish and put ASW in, then pipeted veligers into the tray from the bottom of the bottle. I also pipeted (a different pipet, of course!) Spirulina from one of the culture bottles. It has been growing in a thin layer on the bottom so I was able to get a bit (not a lot, but a nice amounts for this little experiment). I filled it to the tippy top with ASW and a small amount of Iso, and put the cap on it. This is not ideal since there is a noticeable bubble, but the cetyl alcohol that arrived turned out to be chips, not flakes, and it just doesn't seem to work as would be expected. So, petri dish lid it was. I put a dish under the scope to see if I noticed any congregation by the Spirulina. I did not see that, but I only had time to look for about 10 minutes before I needed to move on to another project. This morning when I peeked at the dishes, there were more in the water column than on the bottom...so maybe another water change is needed? Maybe a lower density is needed at this development stage? I have no idea at this point.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Sunday, April 28, 2013 7:31 PM
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So, if I count the day of hatch as Day 1, the veligers from bottle 4-10 should be competent. These are essentially the only veligers left and there don't seem to be too many. I had previously put several in to 4 different Petri dishes with some spirulina, so either they are dying or they are settling. I don't know yet. Today I added some spirulina to the bottle itself since that was easier than transferring everything to Petri dishes. I don't feel I have enough veligers to try other algae species (not that I really have any better options) so fingers crossed that I have some alive and that the spirulina will work!
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, April 29, 2013 6:55 PM
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Good luck! How did the ones you put in the specimen dish on Thurs. do?
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, April 29, 2013 8:24 PM
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I don't think they were competent to settle until yesterday/today...and I don't think I will check until tomorrow. Meta takes at least 24 hours, IIRC. The only noticeable difference about the ones in the dish were that instead of just sitting on the bottom, they went back into the water column. The Spirulina has grown very well in the Petri dishes in the meantime so that's a good thing if they do end up settling on it. Tomorrow I will put some under the scope if I feel brave...they are still less than 300uM so I can't tell what's going on with them by eye.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Tuesday, April 30, 2013 4:24 PM
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Well, I'm rooting for you. I don't know anything about sea hares but I love watching veligers under the microscope.
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Thursday, May 2, 2013 9:22 AM
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So I still haven't had a chance to look at these under the microscope. I have been looking at them in the dish just with my eye, and there are some (in one dish only) that are still in the water column. The other petri dishes don't have any in the water column. They could be dead or they could have settled. The feeling I have right now is that they are dead since there are some swimmers in the one dish yet that clearly have not dropped down on to the substrate. Although, it's possible that some have, and some have not. Previous studies called a substrate a "settlement substrate" if 50% settled. So maybe...just won't know until I peek under the scope since they are still far too small to tell what is what. I don't yet know when I will have a chance to do this (one of my dogs lacerated her chest pretty bad so my time has been spent going to the vet and changing bandages the last few days )
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Tuesday, May 7, 2013 9:28 AM
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Okay, so I FINALLY got around to looking at these closely under the microscope and it was as I had feared (expected too, I suppose) and they were dead. There had been several some dots where spirulina was growing nicely, and those were the spots the dead veligers were. Reading Paige (1988) Table 3 shows a comparison between development of B. leachii and Aplysia. Day 12 is when B. leachii develops a larval heart and Day 15 they reach maximum shell size. It is between days 12 and 15 that the majority of the veligers would drop out of the water column on me (and not to settle, but rather to die it would seem). I am wondering if having good water quality is critical at that stage.... I won't be able to test that until I get some new adults. I would like to get more of the "blue dot" variety (from the Indo-Pacific typically) since I find them to be more attractive, but I would also like to just get some of the "ragged seahare" B. leachii since I think this is the actual kind Paige was working with since they were collected around the coast of Florida. I would also like to perfect the larval rearing containers to do something along the lines of what EasterEggs mentioned previously in this journal. It would just be a matter of finding a tube that fits nicely over my bottles, I think. A bench top roller would be perfect, but would also take up quite a bit of space and would limit the number of containers I could have. Until then, this journal is unfortunately on HOLD. Booooooo
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Tuesday, May 7, 2013 2:27 PM
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Oh, too bad. I was rooting for you. Is spirulina the cue these veligers need in order to settle? I don't know anything about them but inverts can be weird sometimes (well, most of the time). Most inverts won't settle until they have a specific cue in their environment and lots will just die if they never get that cue. But like I say, I don't know anything about sea hares.
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Tuesday, May 7, 2013 2:49 PM
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So a Spirulina species, not the exact one I used, was used and verified as a settlement substrate for B. leachii pleii (which is the Florida/Caribbean subspecies). The blue dot B. leachii (I don't know the exact subspecies of mine) are typically from the Indo-Pacific, so Spirulina species may not work for them. In general, B. leachii seems to need blue-green algaes/cyanobacteria so I was hoping that the Spirulina I was able to get rather easily would work. I still hold out hope that it could, but I obviously can't verify that yet.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Tuesday, May 7, 2013 3:00 PM
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Well, good luck. I hope you get a new chance to work with them soon.
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, January 6, 2014 9:06 AM
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This weekend I picked up two blue dots! I haven't seen any egg strings yet, but I am hoping to see some in the next week or so I can finally get this project going again! I plan to try a couple different things this time around. First, some veligers I think I will just keep in a bubbling water bottle that has been modified a bit (I haven't quite nailed down exactly how I plan to do it, but I will post pictures when I figure it out). I also have this shaker thing I found and bought on eBay for like $20 that I need to modify a bit to keep it going continuously and I want to try some things with that as well. For settlement cues, I bought freeze dried spirulina and I am hoping that will work well enough. Otherwise, I will try to find some green slime algae from my own tanks or from some local reef club members' tanks and use that (I will likely try both).
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, January 6, 2014 12:52 PM
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Cool. Tagging along. I'm anxious to see how this works out. FWIW, I have read that Ulva can be used for settlement cue.
"If everything is going good, you've obviously overlooked something."- Martin Moe, Jr
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, January 6, 2014 2:27 PM
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I vaguely remember reading something about Ulva and settlement, but from what I remembered, it was actually enteromorpha or something, not Ulva...I can't recall precisely and I think there has also been some rearranging of taxonomy that I haven't really been able to follow. I will have to scrounge up my papers on B. leachii again. And maybe I will look into getting a bit of both types of algae, just in case
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Monday, March 10, 2014 8:28 AM
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Well, the other two I bought just up and disappeared, never to be seen again. I have replacements now (three) and put them in a breeder basket so that way it will be much harder for them to just disappear. Yesterday, I found an egg string I will probably pull it out on Tuesday for hatching.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Wednesday, March 12, 2014 8:12 AM
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I had planned to pull it last night...but they had already hatched! Now I have wait for the next egg string, but I'm hoping I won't have to wait long. All three were in a giant blob ball this morning when I checked on them. Hopefully they were spawning.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Wednesday, March 12, 2014 8:26 AM
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Shannpeach, your veliger pics earlier, you notice the bubbles around pic 2 of the close ups? I noticed almost the exact same thing with my cerithium veligers, and then next day or wo 90-100% of them would die off. Did you notice the bubbles moving around when you saw them? (if you remember??) I think they are some sort of veliger parasite.
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Re: Breeding Journal, Species: Bursatella leachii (Blue Dot Sea Hare)
Wednesday, March 12, 2014 8:44 AM
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I'm not sure I see what you are talking about... I have noticed in general, that bubbles/air of any kind is problematic and causes them to die. So I'm not sure I agree that it is a parasite, but think the problem is more likely microbubbles, possibly produced by the live phytoplankton. I have also noticed though that within a day the majority just drop to the bottom and appear to be dead. I'm not sure what the cause is though and don't want to speculate yet. It was often around the time that they should be nearly competent to settle, but it's been awhile so I can't remember exactly. I'm looking forward to trying again though
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