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> > Preservative solution % ratios ????

BrookymanJune 21st, 2012, 3:30 pm
Banned
Posts: 797
Hi Guy's


We all can say we've seen many different ratios Alcohol %.

..In Luke post bug collectors wanted he requested just

in 100 % alcohol.



??? What is the best ratio, that you pro's could agree on ????




Reason I ask . Sometimes I get a bacteria ???

growth on them. I believe that it might be sometime I have

left to much air in the test-tub. What I have started to do

with some of them is. Put them in hydrogen peroxide straight.

Then I add bleach, It works kind of like Vinegar baking soda.

This cleans them perfectly.


However I found out that if you use lot's of bleach It

will clean out the thorax & abdominal cavities digestive

remains and muscle mass which I then mount the shuck

on slides.


It also ( does not remove ) the EXO shucks black stains or amber tones.

As well it seem to not effect the mouth parts or exteriors

like the gills.


Thank You

Mack


Banned for threatening another user and then trying to circumvent a kinder "soft" ban with fake accounts
CrepuscularJune 21st, 2012, 6:06 pm
Boiling Springs, PA

Posts: 919
As high a proof alcohol (Ethanol) as you can get is what you want. If you can get the "real" stuff as in non-denatured that is the best. You will lose colors over time, but it preserves them the best. You can get 95% denatured from places like bioquip. I have access to 100% ethanol but it's a REAL hassel to get. We have to have an actual liquor license here in PA to purchase the stuff. I personally do not like to work with the denatured stuff mainly due to smell. But since you are in CA maybe it will be easier for you. I would not use anything less than 75%. I have samples that are >10 years old in ethanol and they are fine. Make sure you have good vials with a poly seal cap, ethanol evaporates quickly.
There are other old-school solutions out there but they contain some pretty nasty stuff. I do not recommend using them.
EntomanJune 21st, 2012, 7:17 pm
Northern CA & ID

Posts: 2604
Mack -

Ditto what Eric said. I would only add that there are three stages: Initial or observational period and long term. For the first few weeks, don't laugh, but I like gin (preferably Bombay Sapphire). The leftover liquid after transfer makes excellent martinis! Hey, be the trout...:) All kidding aside, there's a practical reason as well. It is in the first couple of weeks that I fiddle with them most for inspection (and now photography). I've found that the harsher concentrations make the specimens more brittle whereas in the gin they remain quite pliable and retain much more of their color. I'm convinced that there's something in the gin that enhances this. Perhaps the juniper berries? I hate losing appendages to handling.

Here's the protocol I follow:

Streamside kill vials - Gin; if not used streamside, used later in the evening. Always go fishing with fresh gin.

Initial inspection period (a few weeks?) - store kill vials in the fridge, pulling out only for photo sessions or inspection. This is important as the gin lacks enough alcohol to prevent spoilage if stored at room temp for very long. The alcohol content drops even more as the bug's bodily fluid goes into solution.

Long term storage (stage 1) - transfer the specimens from the refrigerated gin kill vials to labeled ones filled with min. 70% ethyl alcohol. Move onto stage 2 in a couple of days or put back in the fridge.

Long term storage (Stage 2) - Replace the alcohol from stage 1. For most small specimens (if not overcrowded) a couple of days will insure that the water and soluable fats has been purged from the specimens. For crowds or large specimens (like big stoneflies or caddis larvae) you may need to repeat this stage a few times more.

The key is getting rid of the bug's water. When done properly, I have had excellent results with the lower concentrations (70%). The advantage is more color and flexibility is retained. I've got specimens pushing more than 20 yrs that look to be doing fine, though I admit to reticence in messing with them. I have some specimens that are over ten that I photographed last year. The only damage done was to a specimen I tried to force into a pose (lost a leg).

BTW - I'm not so sure peroxide is a good idea. Though, I'm sure it will remove the broken down insect cells (scum) clinging to your specimens, it will also burn away important body parts as well - particularly gills. I'm sorry, but there's really no way to fix a spoiled specimen.

When it comes to clearing specimens for dissection & slide mounting, I remember Creno offered up some excellent sources for information on this topic in a post a few years back.. I wish I also remembered where it was! Hopefully he'll check in.





"It's not that I find fishing so important, it's just that I find all other endeavors of Man equally unimportant... And not nearly as much fun!" Robert Traver, Anatomy of a Fisherman
CrenoJune 21st, 2012, 8:11 pm
Grants Pass, OR

Posts: 298
Gin huh? Well since I don't like to drink it I guess killing critters for a better cause would be an acceptable use.

The 100% ETOH non-denatured recommendation from Luke is because he is looking for material for DNA analysis. Me too. And 100% preserves them faster in the field as they always come with some water. If I fill more than a third of a vial with critters I always change out the ETOH the next day. And maybe again if they are big fat critters.

I have found that collecting into 100% and changing to 70% non-denatured ETOH after a day or two is fine for reducing brittleness and long term storage. It has been used for decades in museums. And I get pretty good DNA results as long as the critters are processed within 10 years or so. I am also getting good DNA results from critters preserved in the field with rubbing ETOH and then to 70% within a month or so as long as the critters are processed in year or two. But 100%, in combination with refrigeration, is probably the best fisherfolks have for both DNA and your collection/examination.

ETOH does change the colors but as far as I know there are no preservatives that maintain the colors ya see in a live animal. To be expected - death changes alot of things........

As far as clearing specimens - I will try the peroxide and bleach. Anything is worth trying a couple times. So far I have settled on cold 10% potassium hydroxide (KOH). Some folks heat the stuff up a little to speed up the process but it is very caustic and the hot stuff will eat a hole in ya. DO NOT spatter in an eye! Others recommend lactic acid but they heat that as well and I have not had as good results as KOH so not much experience with that.

I have the same trouble as others getting non-denatured ETOH. Ya need to work with a taxonomist someplace that has access. Or maybe a farmer? In the midwest they raise corn just to turn to ETOH. Probably takes more energy than the ETOH produces but as far as I know it is a tax subsidized process so that doesn't really matter. Oh well. I think they use it in farm vehicles so maybe the would give ya a quart of so.

Be careful of denatured ETOH as they can denature it with stuff that is not healthy for ya. They do it for a reason but, depending on what is in it, it may not be good to breath as you pour over those specimens. The same with rubbing ETOH.

creno
CrepuscularJune 21st, 2012, 10:00 pm
Boiling Springs, PA

Posts: 919
Mack -

don't laugh, but I like gin (preferably Bombay Sapphire). The leftover liquid after transfer makes excellent martinis! Hey, be the trout...:)




Don't laugh, yeah right. Nice observations though Kurt, I actually did this last week but I used vodka (my wife was with me :) ) and the color of the flies is still intact. while some others that I put in 100% ethanol a couple days earlier are starting to fade. I wonder what it is in he distilled spirits that prevents the color from fading. I'm sure it is not preserving them in the way that the ethanol does. My Bombay Sapphire goes from freezer to glass, co-mingles with some olives or onions and then to mouth, it has never traveled streamside. I have a friend who at one time specialized in Chironomid identification and I have his method for clearing head capsules around somewhere. I'll try and find some time to look for it.


CrepuscularJune 21st, 2012, 10:13 pm
Boiling Springs, PA

Posts: 919

Be careful of denatured ETOH as they can denature it with stuff that is not healthy for ya. They do it for a reason but, depending on what is in it, it may not be good to breath as you pour over those specimens. The same with rubbing ETOH.

creno

Dave thank you for elaborating on the "nature" of denatured ethanol, we had a bunch of old raw stream samples preserved in denatured booze that we had a really hard time getting rid of due to all of the "other stuff" that they use to denature ethanol. It was classified as hazardous waste, and therefore was a bugger to dispose of. I think this is important to point out to Mack because it sounds like he is doing a lot of this stuff with his son. That's why I alluded to fixatives like Kahle's solution etc. In my opinion there is no need for them especially now that I know that not only can I pickle myself with gin, I can also pickle some bugs. :)
BrookymanJune 21st, 2012, 10:44 pm
Banned
Posts: 797
I new I could get good answers. I have been use 70% rubbing but here is
another interesting one I am experimenting we right now.


1 : Citric Acid APX 1/2 teaspoon the rest water In plastic test tube

2 : citric Acid 1/2 teaspoon the rest my typical 70/30 ACH

3 : just my 70/30

All are the same species of larger adult female stone flies with eggs.

I started this test about oh a week and a half ago. So far #1 is winning
her color matches the photo still. There no growth and I have not change
any of them they are in what they died in.


I love Gin & sprite so you know where that's going,,,down the "hatch"..

But I will try it sounds great.

BTW / I thought of the citric acid cause 1 we preserve

food with it. And 2 is a concept that blew me away..

I lived in FT Lauderdale for 10 years. That area has
more termites and ants than it has sand. I met a guy's
that own his own extermination company.

For years he was developing a new method that would
not hurt anything but the bugs. And environmental
friendly + smells great.


It was Orange oil, it burns there skin and internals
so they were dead it 30 ish seconds. My old cottage had
more terms than wood.

Just injecting Orange juice in the canals did the job..
One would be coming out of a hole and die, that plugged
the hole and the all rest behind him died.

I am watch I hope we I open them that the acid+h2o
is the winner. So far its in the led..




Mack.
Banned for threatening another user and then trying to circumvent a kinder "soft" ban with fake accounts
BrookymanJune 21st, 2012, 11:45 pm
Banned
Posts: 797
Just thing Kurt..

Have you tried just allowing a sample to air dry then
put in solution...Oh that does nothing RE: the fat.

Cause I re-hydrate adult in water for dissection all the time.

Just a thought.

Mack
Banned for threatening another user and then trying to circumvent a kinder "soft" ban with fake accounts
EntomanJune 22nd, 2012, 12:07 am
Northern CA & ID

Posts: 2604
Mack -

No I haven't tried that. I don't mess with clearing and dissection. I'll leave that to the professional taxonomists who are properly trained and with the necessary skills for such tasks. I just piddle around with my own personal collections for low magnification observance and photos. If I can't figure out what it is, I'll send it off to someone more learned who takes pity on me and hope for the best.:)
"It's not that I find fishing so important, it's just that I find all other endeavors of Man equally unimportant... And not nearly as much fun!" Robert Traver, Anatomy of a Fisherman
BrookymanJune 22nd, 2012, 12:32 am
Banned
Posts: 797
Kurt

I am enjoying look at that mic stuff, and I have learned
so much it only gets real tuff at the very last keys like
counting satae hairs and sockets at 400x. Its like solving
a mystery. Up till this year it been ahhhhh subvaria.

Funny thing I have read more in the last 3 months than I
in the last 10 years.


I fell into the taxo stuff cause I have been fumbling around
with a thesis for a book I started in 1982. In part,
understanding the muscle development and thier complete
anatomy Is a part of the understanding of
cause and effect of my theory.

I have been on Quantum physic, M theory string theory, and more.
They are all metamorphic-ly tangled up inside the entomology body development and the waters pressures.

In short I may at the end have found the ultimate and complete
explanation of everything through the little creators.


Mack
Banned for threatening another user and then trying to circumvent a kinder "soft" ban with fake accounts
FalsiflyJune 22nd, 2012, 2:13 pm
Hayward, WI.

Posts: 656
I have been on Quantum physic, M theory string theory, and more.
They are all metamorphic-ly tangled up inside the entomology body development and the waters pressures.

In short I may at the end have found the ultimate and complete
explanation of everything through the little creators.


Good, maybe then someone can explain how a butterfly flapping its wings in Brazil determines my fly selection.

Falsifly
When asked what I just caught that monster on I showed him. He put on his magnifiers and said, "I can't believe they can see that."
BrookymanJune 22nd, 2012, 7:04 pm
Banned
Posts: 797
Kurt

As well you are the organized detail oriented type.
With the knowledge on this sight you should take a
crack at it. You would be very good at it anyway.
Look at all I have learn in the past six month
or so. But if you do on the other hand, you get swallowed
by trying your best to solve the mystery at hand.

Mack.
Banned for threatening another user and then trying to circumvent a kinder "soft" ban with fake accounts

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