[Nhcoll-l] Alcohol concentration for terrestrial vertebrates
Dirk Neumann
neumann at snsb.de
Fri May 7 09:43:11 EDT 2021
Dear Mare,
are you sure that this is only ethanol, and not something like Kew-mix
or a similar mixture? I am wondering because the EtOH concentration is
rather low, which might be an indication that further ingredients might
be included in this preservation fluid.
Originally, Kew-mixture was 53% EtOH, 37% water, 5% formaldehyde, and 5%
glycerol. As far as I know, the formula was changed to 70% ETOH, 29%
water, and 1% glycerol in the 2nd edition of The Herbarium Handbook
(1989). But I am not a botanists, and others might be in a better
position for giving advice?
Strong dehydration of cells can be an issue, this is why the glycerol is
added.
With best wishes
Dirk
Am 07.05.2021 um 14:53 schrieb Mare Nazaire:
> This is a very informative and helpful thread - thank you for this!
>
> I presume that 70% concentration would also be suitable for plant
> material preserved in spirits? I ask because I've recently discovered
> that some of our collection of fluid preserved plant material is at a
> concentration of 50% and I wondered if it is advisable to keep them as
> is or change their concentration to 70%. Are there recommendations in
> John Simmon's book for preserving plant specimens in alcohol and could
> you also provide the citation for this book?
>
> Thank you,
> ~Mare
>
> On Fri, May 7, 2021 at 12:48 AM Erik Åhlander <Erik.Ahlander at nrm.se
> <mailto:Erik.Ahlander at nrm.se>> wrote:
>
> Dear Tonya, John, Simon, Dirk - well all,
>
> Also I agree. Since I will soon retire I want to share some
> experiences:
>
> When we started to take care of the collection of wet vertebrates
> in Stockholm in 1975 there was no overlap in time (well 1 week)
> with the previous staff (the previous curator was employed
> 1934-1974). So we had to invent the wheel. The initial ambition
> was to keep a concentration between 70 and 80% ethanol. (We also
> tested the new suggested conservation fluid Phenoxetol, which
> after some years showed to be a disaster). To compensate for
> evaporation, we tried to stick to 80%. New material was fixed in
> formalin for at least a week, washing in water, 20% ethanol for
> two days or more, 50% for two days or more, and final storage in
> 80%. Also we removed all bad jars from the collection – and a bad
> jar was a jar that needed topping. Expedition material was sorted
> and identified etc after this stage with the result that many
> specimens was changed to 80% once more. It took more than 10 years
> to realize that 80% was to strong. But also that every change of
> alcohol, or topping, resulted in a higher concentration ethanol
> since the lowering effect of the alcohol concentration through
> remnants of the previus stage fluid inside the specimens was
> removed. Also the small amounts of formalin in the specimen was
> reduced for each change of fluid. Especially for tiny fish we
> could find obvious shrinking. Today we are careful
>
> 1.To keep the specimens in 70% (not more, not less)
>
> 2.Not to rinse to much in water. Rather remove the formalin from
> the surface of the specimen only.
>
> 3.Don´t change the fluid if it is not necessary.
>
> 4.If you have to remove all fluid, add maybe 80-90% of fresh (70%)
> ethanol and the rest used ethanol from another specimen.
>
> All formalin fixed specimens has a small amount of formalin left -
> that is good.
>
> Some substances in the specimen dissolve in the alcohol (just look
> at an alcohol preserved Anguilla…). Every change of alcohol add to
> the removing of lipids etc - that is bad.
>
> As far as we know, formalin was used for the first time at the NRM
> in 1904, but only occasionally! Still in the 1940s ethanol was
> commonly used for fixation in the field. When the museum moved
> from downtown Stockholm to north of the city in 1916, the economy
> for alcohol was reduced due to world war I (otherwise Sweden was
> not involved). This led to the invention to use a diluted formalin
> solution for the exhibition jars (for specimens fixed in
> ethanol!). The research collection continued to be stored in
> ethanol. Our collection is old. We estimate that our oldest
> specimens in ethanol are from the 1720s (from the Seba
> collection). Still many specimens from before 1758 are in
> remarkable good condition. In some specimens it is even possible
> to get small pieces of DNA with ancient DNA technic – but usually
> not. This sounds contradicting to some statements above. We don’t
> know too much about the preservation history of these specimens,
> but what we know might be of general interest. The initial
> fixation and preservation was in distilled wine (=“spiritus
> vini”). We don’t know the concentration, and probably it was not
> pure ethanol, but also contained small amount of other fractions
> from the wine, more like strong cognac. The Royal collection (of
> king Adolf Fredrik with many Linnaean types) was donated to the
> Royal Swedish Academy of Sciences in 1801. NRM was founded in
> 1819, but immediately in practice fused with the Academy. In 1848
> the collections of the Academy was formally donated to the Museum.
> From the 1740s to 1970 this collection of vertebrates in alcohol
> was moved four times. Jars and fluid was probably changed twice.
> But most of the time the collection was stored cool and dark.
> Glasses and fluids was expensive so the ratio: specimen volume /
> conservation fluid volume was high up to 1900. From 1801-1898 the
> major part seems to have been almost untouched, except that the
> whole collection was moved 1500 meters in 1829.
>
> I was once asked how long a specimen could be stored in alcohol.
> With the reservation that our old specimens will be stored like
> today, no sudden disasters etc (and no climate change), I decided
> that to 2220 = 500 years would be possible, maybe 1000 years.
>
> Erik Åhlander
>
> vertebrate zoology and museum history
>
> ZOO
>
> Swedish Museum of Natural History
>
> PO Box 50007
>
> SE-10405 Stockholm
>
> Sweden
>
> +46 0 8 5195 4118
>
> +46 0 70 225 2716
>
> erik.ahlander at nrm.se <mailto:erik.ahlander at nrm.se>
>
> *Från:*Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu
> <mailto:nhcoll-l-bounces at mailman.yale.edu>> *För *Dirk Neumann
> *Skickat:* den 7 maj 2021 08:36
> *Till:* nhcoll-l at mailman.yale.edu <mailto:nhcoll-l at mailman.yale.edu>
> *Ämne:* Re: [Nhcoll-l] Alcohol concentration for terrestrial
> vertebrates
>
> Hi Tonya (and John and Simon ;-)
>
> concur with John and Simon, specimens should be kept in 70%; Simon
> pointed to the diluting effects and the image below nicely
> illustrates this: even if you use more steps for transferring
> specimens (0/20/40/60/80 vs. 20/30/50/70), tissues are still
> soaked with 60% or less high concentrated EtOH.
>
> Depending on size, body mass and number of specimens (i.e. amount
> of tissue in the jar), the effect can be considerable (see
> "staining" in the images below; in the left one, body fluids
> released from these tall whitefish are indicated by the reddish
> haemoglobin stain at the bottom of the jar, the overall greenish
> colour in the right comes from chlorophyll released from the guts
> of these herbivorous distichodus fish).
>
> I do the initial filling usually with 73-75% EtOH to reach 70%;
> aside from vertebrates high EtOH concentrations can be an issue in
> malaise traps because there the specimens usually are collected
> over several days or weeks in 96-80% EtOH. As Simon pointed out
> this quickly dehydrates specimens and weakens the joints holding
> all the antennae, appendices, bristles of invertebrates. Another
> issue is that in unsorted malaise trap samples there often is a
> thick deposit of specimens at the bottom of the container. Because
> the diluted less high concentrated ethanol is heavier, it layers
> at the bottom of the jar (cf. whitefish jar). Inside malaise trap
> containers, this diluted EtOH may get trapped in the thick
> specimen deposit.
>
> Usually, I leave jars for few day to see if there are any unwanted
> effects before moving them into the collection.
>
> Hope this is useful, with best wishes
>
> Dirk
>
> Am 07.05.2021 um 00:17 schrieb Simon Moore:
>
> Thanks John and Tonya,
>
> What John says is true about the staging of alcohols and the
> final concentrations. 80% was what I was advised at the NHM
> in London when I worked there and by the time larger
> terrestrial vertebrates ‘end up’ in 80%, you will often find
> that with the mix of lower grade alcohols from the staging
> process, once things have settled down / equilibrated, then
> the net result is around 70% anyway. Higher grade alcohols
> can lead to embrittlement of certain tissues as well as
> evaporation issues.
>
> I have also found the staging process necessary for the more
> fragile specimens as they undergo changes in Osmotic pressure
> during this process which can cause syneresis or shrinkage in
> softer tissues.
>
> With all good wishes, Simon
>
> Simon Moore MIScT, RSci, FLS, ACR
> Conservator of Natural Sciences and Cutlery Historian,
>
> www.natural-history-conservation.com
> <https://url11.mailanyone.net/v1/?m=1leu5X-0006gs-5g&i=57e1b682&c=P_JTOxhc00RtGTsMjTrryrRBThMnzI5k1ol2aDVqPITm0G_xR0drpuNIhh-krJ6ihFhOLJnXYNjI5fJDeS7rag0t-LwIYs0jmRWXIk2uN2sYVvoo4O9RHPsKEAKiAK-LvbrlH-pnEJMM5dJlJOlNvswIXfaiFJxHBKwsoJX5uQ31zmivYbvBNJdb61ZNkqDKGinISZmQBmu6t6VBola0IT4zHh0nqkiHmWvI7KCXEbWncO8-owQTcerGpMed6sP9>
>
>
>
>
> On 6 May 2021, at 22:50, John E Simmons
> <simmons.johne at gmail.com <mailto:simmons.johne at gmail.com>>
> wrote:
>
> Tonya,
> Thank you for your kind words about my book. The
> recommendation for staging up to 80% concentration was by
> made by my friend Simon Moore, who I cited in that
> sentence. In general, I do not recommend using 80% ETOH as
> a preservative for terrestrial vertebrates, but rather
> 70%. Preservation is alcohol is a trade-off between
> dehydration of the specimens and providing them suitable
> protection against biological deterioration. At 70%, ETOH
> is a very good biocide; below that, not so good, and above
> 70%, too strong for most specimens (note that there are
> some instances in which 80% might be preferred).
>
> I do not recommend using stronger alcohol as a hedge
> against evaporation--that leads to uneven concentrations
> of preservatives and can be a real mess to work with in
> a collection.
>
> For how-to instructions on preserving, transferring
> specimens, and managing a fluid preserved collection, you
> might want to check Herpetological Collecting and
> Collections Management (3rd edition, 2015). The
> instructions for preserving and managing fluid preserved
> animals will work for most other specimens as well as for
> reptiles and amphibians.
>
> Hope this helps,
> --John
>
> John E. Simmons
> Writer and Museum Consultant
> Museologica
> and
> Associate Curator of Collections
> Earth and Mineral Science Museum & Art Gallery
> Penn State University
> and
> Investigador Asociado, Departamento de Ornitologia
> Museo de Historia Natural, Universidad Nacional Mayor de
> San Marcos, Lima
>
>
> On Thu, May 6, 2021 at 4:05 PM Haff, Tonya (NCMI, Crace)
> <Tonya.Haff at csiro.au <mailto:Tonya.Haff at csiro.au>> wrote:
> Hello all,
>
> I am enjoying reading John Simmon's fantastic book on
> fluid preservation. In it I read one suggestion for
> stepping specimens up out of formalin fixative
> into preservation alcohol as follows: from 20% ETOH to 40%
> to 60% and finally to 80%. We typically place our
> specimens in 70% ETOH, and I know higher concentrations
> can cause some problems with specimen dehydration. All our
> specimens are terrestrial vertebrates. I presume the final
> 80% provides a buffer against ETOH evaporation or leaching
> of water from the specimen into the fluid in the jar, to
> ensure that the alcohol concentration in the preservation
> fluid stays sufficiently high? But to me this is not quite
> clear. I wonder if any of you have thoughts on this, or if
> you would be willing to share how you step
> your specimens up in ETOH?
>
> Thank you!
>
> Tonya
>
>
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>
>
> Dirk Neumann
>
> Tel: 089 / 8107-111
> Fax: 089 / 8107-300
> neumann(a)snsb.de <http://snsb.de>
>
> Postanschrift:
>
> Staatliche Naturwissenschaftliche Sammlungen Bayerns
> Zoologische Staatssammlung München
> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
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>
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> ---------
>
> Dirk Neumann
>
> Tel: +49-89-8107-111
> Fax: +49-89-8107-300
> neumann(a)snsb.de <http://snsb.de>
>
> postal address:
>
> Bavarian Natural History Collections
> The Bavarian State Collection of Zoology
> Dirk Neumann, Section Ichthyology / DNA-Storage
> Muenchhausenstr. 21
> 81247 Munich (Germany)
>
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> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
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> 909.625.8767 ext. 268
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Dirk Neumann
Tel: 089 / 8107-111
Fax: 089 / 8107-300
neumann(a)snsb.de
Postanschrift:
Staatliche Naturwissenschaftliche Sammlungen Bayerns
Zoologische Staatssammlung München
Dirk Neumann, Sektion Ichthyologie / DNA-Storage
Münchhausenstr. 21
81247 München
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---------
Dirk Neumann
Tel: +49-89-8107-111
Fax: +49-89-8107-300
neumann(a)snsb.de
postal address:
Bavarian Natural History Collections
The Bavarian State Collection of Zoology
Dirk Neumann, Section Ichthyology / DNA-Storage
Muenchhausenstr. 21
81247 Munich (Germany)
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