When they die (and they do)

G

greg

Guest
I was wondering what most folks do when their caudates die. I ask mainly because most people are probably not aware that the species you keep might actually be of interest to your local museum of natural history or to a biology teacher or something. This may strike an interesting emotional cord with some people, and I fully understand that. But.... for those who don't want the end of the animal's life to be the end of the good it can do, there are alternatives. A salamander that is no longer able to offer its life and its behavior etc. can still teach others a great deal.

For those that breed caudates, this is especially of importance because many (most) of the newts and other species I know we all keep and try to breed are rather poorly represented in museum larval collections. That is, if a researcher needed to see what a hind limb-bud stage Cynops orientalis looked like, they'd have a VERY hard time finding that in any museum collection. Yet, that's a stage when caudates do often die during metamorphosis.

All I'm trying to say is WHEN captive caudates die, I hope people will remember that the animal still has a lot to offer the "community" at large. It's value has not ended simply because the life has. If anyone is interested in knowing more about this, please let me know.

Thanks and I hope this doesn't weird anybody out too much. I'm not at all suggesting that people should send their live animals to a lab. I'm just suggesting an alternative for when the critters leave this mortal coil on their own. It's a fact... even if we don't wish it to happen, they eventually will die. What happens after that is up to each of us.

Greg
 
Hi Greg,
very interesting topic. I used to dump the dead animals and only in recent years have started to preserve some in alcohol. Why don't you specify the state the animals should be in as well as how to prepare/preserve them for further use.
I think Kai also posted some information on preservation of dead caudates on this forum before.
Some animals I bought from dealers (I have to admit I sometimes still do) were in a pitiful state before as well as after their death and probably not really worth preserving. I agree though, that once in a while an animal will die without any sign of illness. These are the animals which might especially be useful for further scientific or educational purpose.
As far as "collections" of specimen are concerned though, I have experienced that they are always only as "good" as the person who collects (meaning a collection has to be well organized to be useful).

Ralf
 
I cannot speak for every museum, but I will basically accept any "quality" of specimen and will typically find some use for it. Only in rare circumstances do I dispose of something and then only if it is a really common species that is well-represented in our collection AND in horrible shape.

Thin animals from pet stores (such as the ones who keep Salamandra under hot lamps) are still useful as skeletal specimens.

For me personally I think that putting a specimen into a museum or teaching collection after its death adds to the value of the life it lived. I feel this way especially in cases of wild caught animals. they were taken from wild populations for commercial reasons. What better way to try and right a wrong than by teaching the next generation of students to appreciate these critters. Also, in terms of research specimens, in far too many cases researchers cannot get collecting permits for certain countries, but commercial ventures CAN. So... the only way for some of us to get research specimens are through the trade. Given that I think we all agree that over-collecting in places like Ukraine is a conservation issue of importance, it would be a shame to risk local or complete extinction of species and NOT have them represented in collections.

If there is interest, I can provide specifics on HOW to preserve specimens for the widest possible end-use. I won't clutter this space though if folks are not interested in hearing details.

I will suggest though that people make personal contact with local biology teachers at high schools and universities and see if they have any specific interest. Also, whatever museum of natural history in your area might have interest. In each case there may be specific requirements in terms of not only how they are preserved, but in terms of legalities. For example, we require donors to sign a "deed of gift" form indicating that they won't be wanting the specimen back and giving us permission to use our best judgement with regards to it.

Greg
 
Greg, I am interested in how to initially to preserve a specimen. If the occasion should arise, I'd like to know what to do. Pharmacies sell 70% ethyl alcohol, would that be suitable for temporary preservation?
 
Yes, salamanders tend to go bad very quickly after they die so this would be useful information Greg, so people could have the right stuff on hand/at the ready.
 
Okay. Here are some recipes.

First, the alcohol at most pharmacies in the U.S. is denatured or it is isopropyl. Neither is really good for preserving. Denatured alcohol may have any of over 50 different compounds in it to make it lethal if ingested by humans. Some of these do things like darken or harden or shrivel or CLEAR specimens. I used over-the-counter denatured alcohol once. I kept a metamorph Rana clamitans in it for 3 years and when I recently found the specimen again it appeared to be an albino!... and transparent in some places!

Best alcohol is grain alcohol from your local liquor store. Around here, there is a brand that is ironically called "Grave's Grain Alcohol". It's 95% pure ethanol. I can't imagine anyone drinking it... but it's not denatured. DILUTE THIS. 7 parts alcohol to 3 parts water. Don't worry about the math involved beyond that.

This will work for storage of specimens and will preserve most specimens up to, say, Triturus size without any mechanical requirements. For things larger, like tigers... injection or something is needed to make sure the preservative reaches the core. The good thing about ethanol is that a specimen stored in it can be used as a source of tissue for DNA work, as well as a preserved specimen or a skeletal specimen. There are things it cannot be used for, but that's okay.

For larvae, however, ethanol is not suggested. Alcohol dehydrates, even at 70%. Larval characters such as balancers, gill structure, costal grooves, etc. get lost. In fact, species that do not have costal grooves as larvae will get them if stored in ethanol. The best thing to use for them is a 10% formalin solution. In some places, formaldehyde can be gotten at your local agriculture store. Or from a vet. In some places on Earth I've actually found it on the shelf next to things like foot powder . Formaldehyde and formalin are essentially the same thing except that formalin is diluted and buffered. Formaldehyde usually comes as a liquid and 33-42%. Treat this asif it is 100% formaldehyde. dilute 9 parts water to 1 part that stuff. do not drink it... unlike Grave's Grain alcohol... bad things will happen. for short-term don't worry about the pH or buffering. if you don't know when you'll be able to put the specimens anywhere... buffer with a bit of baking soda. Not too much. if you have a pH test kit for your aquarium... pH should be 7.0. Pure formaldehyde is closer to 4.0.... and dissolves bone on the specimens.

If you don't feel like playing chemistry.... put them in some water and freeze them. Water prevents freezer burn. Freezing is notoriously bad for amphibian specimens though... ice crystals break down membranes and things get very useless for many techniques, including DNA use, histology, etc. but skeletons can still work. Don't freeze larvae though. ice crystals bad to small structures.

So, in a nutshell... Formalin and ethanol.

A very important thing to remember.... put some sort of note inside the bottle with the specimen otherwise you are likely to forget what it is, when it died, etc. Best to use pencil as graphite is alcohol and formalin-proof. Most commonly available inks are NOT and laser printers tend to not be alcohol proof.. Use a heavy card-stock paper so it doesn't break down instantly into a "spit-wad" in the fluid. Index cards work okay.

If you live near where the specimen will be going, and it you can get it there within 48 hours. Refrigerate it (not freeze) and take it to the person un-preserved. That way they can decide from almost all of the options exactly how they want to prep it.


greg

P.S. I've seen some old specimens in the collection that were actually preserved and stored in ETHER. It worked rather well... but scares the heck out of me. I mean, who has access to ether on a regular basis and why?.. the jars weren't even labeled and when we opened them... whew.... we almost blacked out!
 
Re: When they die (and they do)

I realize this is an oooold post, but I wanted to add that I've always used a 10% buffered formalin (BF) solution on adults as well. What we typically do in our lab:

Find a tupperware that will only be used for the purposes of preservation. Place a few sheets of thin hardware or some other cloth (I think white handkerchief cloth would work better for small amphibians) in it. You want a thin cloth so the weight of the wet cloth doesn't flatten your specimen. Pour enough 10% BF to soak the cloth as well as have a about half a centimeter of liquid in the container. Place your amphibian on the 2nd highest sheet of cloth, position as desired, and cover gently with top sheet of cloth. Leave them in overnight or for less time, and then plop them in 70% ethyl alcohol.

What is great about this method is that it allows you to position the animal, and the formalin will ensure that it remains a good quality specimen for longer than if it was just in alcohol. If you have a larger specimen (anything that is 2cm thick or more), you'll need to inject it with formalin, but you can still use this method. This also works for small reptiles.

As a biologist, this is what I have done with all of my amphibians wards that have passed, provided that they weren't decomposing when I found them. If you don't have time to preserve the specimen right off the bat, you can place it in the freezer. But, as previously mentioned, this will reduce it's utility. However, it can still produce a decent specimen for species identification.
 
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Re: When they die (and they do)

What lamb says is probably the best way to keep specimens. However, if you want to do any DNA work with them, don't let them touch formalin. It will screw things up.

Mike
 
Re: When they die (and they do)

Good point, Mike. Forgot to add that. For DNA I think it's best to put them in 90%. When we were looking the DNA of a parasites in the livers of tadpoles that is the concentration we used.
 
Re: When they die (and they do)

The higher the better. Freezing is the best though.

Are you extracting DNA out of those parasite yourself?

Whos lab do you work in??

Mike
 
Re: When they die (and they do)

Good point, Mike. Forgot to add that. For DNA I think it's best to put them in 90%. When we were looking the DNA of a parasites in the livers of tadpoles that is the concentration we used.
Click to expand...

90% of what? I would like to safe all my dead Paramesotriton guangxiensis larvae they will surely die when they hatch, due to them be such rare species.

Let me know how to.
Cheers,
 
Re: When they die (and they do)

eljorgo said:
90% of what? I would like to safe all my dead Paramesotriton guangxiensis larvae they will surely die when they hatch, due to them be such rare species.

Let me know how to.
Cheers,
Click to expand...
90% ethanol. I was also told that for DNA work you want to use alcohol that does not contain denaturants. Thus, if you don't have access to lab-grade ethanol, use high-proof drinking alcohol, not rubbing alcohol.
 
Re: When they die (and they do)

Jennewt said:
Thus, if you don't have access to lab-grade ethanol, use high-proof drinking alcohol, not rubbing alcohol.
Click to expand...

would a newt pickled in high-proof drinking alcohol be...pissed as a newt? :dizzy:


ya ya, one shouldn't joke about such things...:yin-yang:
 
Re: When they die (and they do)

Nice topic! I didn't know anybody, hwo do it, expecting me. And I felt myself so rude, therefore. In the near future I saved some eggs (maybe Rana dalmatina), from a pool, before it ran dry. I bring up them until metamorphosis, and then I set them free in a larger water near the pool, but some of them unforunately died in different stages of evolutionary. I got them into 96% of ethenol. And it's preserved them well. Thaks for informations! And nice to hear that I'm not alone with this idea! (Sorry, my English is cheapness... :D)
 
Re: When they die (and they do)

"In the near future"

I meant bygone... sorry
 
Re: When they die (and they do)

Msteffen said:
The higher the better. Freezing is the best though.

Are you extracting DNA out of those parasite yourself?

Whos lab do you work in??

Mike
Click to expand...


Mike,
No, I wasn't doing the extraction or PCR. I was studying histology slides of tadpoles to determine how the disease progressed and the different stages of the life cycle of the parasite. This was in Dr. Robin Overstreet's lab. He is a well known parasitologist who works at the Gulf Coast Research Lab in Ocean Springs, MS. Why? Are you in a similar field?
 
Re: When they die (and they do)

Lamb said:
Mike,
No, I wasn't doing the extraction or PCR. I was studying histology slides of tadpoles to determine how the disease progressed and the different stages of the life cycle of the parasite. This was in Dr. Robin Overstreet's lab. He is a well known parasitologist who works at the Gulf Coast Research Lab in Ocean Springs, MS. Why? Are you in a similar field?
Click to expand...


Sorta. I am a PhD student in Dr. Ron Bonetts lab at the University of Tulsa. I do mostly phylogenetic stuff and am dabbling into some gene expression stuff.

I was curious about amphibian parasitologists. There arn't very many. We have done a couple projects with amphibian parasites and done DNA extractions with tiny monogeneans and I feel like i should have better success than I do.

Mike
 
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