[Nhcoll-l] Nhcoll-l Digest, Vol 108, Issue 4

Wed May 12 14:09:57 EDT 2021

Hi!

Is there a good reference showing isopropyl not suitable for DNA conservation?

Best regards
Lennart Lennuk
Head of collections
Estonian Museum of Natural History
(+372) 6603404, 56569916

From: Nhcoll-l [mailto:nhcoll-l-bounces at mailman.yale.edu] On Behalf Of Camm Swift
Sent: Wednesday, May 12, 2021 7:54 PM
To: nhcoll-l at mailman.yale.edu
Subject: Re: [Nhcoll-l] Nhcoll-l Digest, Vol 108, Issue 4

Very interesting and comprehensive accounts of alcohol use.  Perhaps covered in Simmon's book, but in the USA, 1950s-1970s at least, considerable use of isopropyl alcohol occurred with fish collections often having jars of both and intentional and inadvertent mixing and/or replacement of one with the other.  Usually dispensing with the isopropyl and replacing with ethanol.  Even collections sticking with ethanol could be faced with taking on orphaned collections in ispropyl so it is something to be dealt with.  I believe since the adverse effects on histology and DNA by isopropyl came to light few if any collections still use isopropyl but it is a strong historical presence in some collections even if just as a residual.  Camm Swift

On Wednesday, May 12, 2021, 12:37:23 AM EDT, nhcoll-l-request at mailman.yale.edu<mailto:nhcoll-l-request at mailman.yale.edu> <nhcoll-l-request at mailman.yale.edu<mailto:nhcoll-l-request at mailman.yale.edu>> wrote:

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Today's Topics:

  1. Re: Alcohol concentration for terrestrial vertebrates
      (Simon Moore)
  2. Re: Alcohol concentration for terrestrial vertebrates
      (Mare Nazaire)
  3. Re: Alcohol concentration for terrestrial vertebrates
      (John E Simmons)
  4. Re: Alcohol concentration for terrestrial vertebrates
      (Mare Nazaire)
  5. Re: Alcohol concentration for terrestrial vertebrates
      (John E Simmons)
  6. Re: Alcohol concentration for terrestrial vertebrates
      (Simon Moore)
  7. Re: Alcohol concentration for terrestrial vertebrates
      (Dirk Neumann)
  8. AIBS Writing for Impact and Influence Online Course: Register
      Now (Jyotsna Pandey)
  9. Alcohol concentrations for preserving flowers (Gretchen Meier)
  10. SPNHC opening mixer (Bentley, Andrew Charles)
  11. Re: Alcohol concentrations for preserving flowers (Simon Moore)
  12. Head of Conservation Unit at the Natural History Museum of
      Denmark (NHMD) (Nikolaj Scharff)
  13. Re: Alcohol concentrations for preserving flowers (John E Simmons)
  14. Re: Alcohol concentrations for preserving flowers (Dirk Neumann)
  15. SPNHC Education DemoCamp 2021 (membership at spnhc.org<mailto:membership at spnhc.org>)
  16. freezing blown eggs (Haff, Tonya (NCMI, Crace))

----------------------------------------------------------------------

Message: 1
Date: Fri, 7 May 2021 15:18:16 +0100
From: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
To: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>, Dirk Neumann <neumann at snsb.de<mailto:neumann at snsb.de>>
Cc: Erik ?hlander <Erik.Ahlander at nrm.se<mailto:Erik.Ahlander at nrm.se>>, NHCOLL-new
    <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID: <945E53CC-DBBD-41D2-8A43-3CC71B88F60A at btinternet.com<mailto:945E53CC-DBBD-41D2-8A43-3CC71B88F60A at btinternet.com>>
Content-Type: text/plain; charset="utf-8"

Dear Mare,

I have always fixed fresh plant material in Kew mix and then transferred to Copenhagen mixture which is similar but minus the formalin.  As Dirk has pointed out, the formulae (proportions) do vary slightly between institutions, some prefer more glycerine in their mixes but which can make the specimens rather translucent which is why others prefer a lower concentration.  There is also the slight problem of osmotic pressure differential and specimens floating until they equilibrate!
Make sure that the pH of the solutions is a near to 7.0 as possible.

With all good wishes, Simon

Simon Moore MIScT, RSci, FLS, ACR
Conservator of Natural Sciences and Cutlery Historian,

www.natural-history-conservation.com<http://www.natural-history-conservation.com>

> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>
> 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 ?hlander <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 i
nitial 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 ?hlander
>
> 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>
>
>
>
>
>
>
>
>
>
> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
> Skickat: den 7 maj 2021 08:36
> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
> ?mne: 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
>
>
>
>
>
>
>
> <image001.png>
>
>
>
> 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<http://www.natural-history-conservation.com>
>
>
> <image002.jpg>
>
>
>
>
> 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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> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
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> natural history collections to ensure their continuing value to
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> Advertising on NH-COLL-L is inappropriate.
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> --
>
> <image004.png>
>
>
> Dirk Neumann
>
> Tel: 089 / 8107-111
> Fax: 089 / 8107-300
> neumann(a)snsb.de
>
> Postanschrift:
>
> Staatliche Naturwissenschaftliche Sammlungen Bayerns
> Zoologische Staatssammlung M?nchen
> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
> M?nchhausenstr. 21
> 81247 M?nchen
>
> Besuchen Sie unsere Sammlung:
> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>
> ---------
>
> 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)
>
> Visit our section at:
> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>
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> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268
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> Advertising on NH-COLL-L is inappropriate.

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------------------------------

Message: 2
Date: Fri, 7 May 2021 07:55:58 -0700
From: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>
To: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
Cc: Dirk Neumann <neumann at snsb.de<mailto:neumann at snsb.de>>, Erik ?hlander
    <Erik.Ahlander at nrm.se<mailto:Erik.Ahlander at nrm.se>>, NHCOLL-new <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID:
    <CAGsrPo+cGTyuw3wsBDqQ3Z5cameTn0k_40Fui4H1j6c84F3F_w at mail.gmail.com<mailto:cGTyuw3wsBDqQ3Z5cameTn0k_40Fui4H1j6c84F3F_w at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Thank you Simon and Dirk for your feedback on this.

I had actually spoken with the botanist who originally prepared the
specimens back in the 60's - he noted that they were preserved in 50% EtOH.
He also noted that when he had traveled to other countries for field work
and EtOH wasn't available he would use rum! So there could be some other
residual components in these fluid preserved specimens!

On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
wrote:

> Dear Mare,
>
> I have always fixed fresh plant material in Kew mix and then transferred
> to Copenhagen mixture which is similar but minus the formalin.  As Dirk has
> pointed out, the formulae (proportions) do vary slightly between
> institutions, some prefer more glycerine in their mixes but which can make
> the specimens rather translucent which is why others prefer a lower
> concentration.  There is also the slight problem of osmotic pressure
> differential and specimens floating until they equilibrate!
> Make sure that the pH of the solutions is a near to 7.0 as possible.
>
> With all good wishes, Simon
>
> Simon Moore MIScT, RSci, FLS, ACR
> Conservator of Natural Sciences and Cutlery Historian,
>
> www.natural-history-conservation.com<http://www.natural-history-conservation.com>
>
>
>
>
> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>
> 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 ?hlander <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 ?hlander
>
> 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>
>
>
>
>
>
>
>
>
>
> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
> Skickat: den 7 maj 2021 08:36
> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
> ?mne: 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
>
>
>
>
>
>
>
> <image001.png>
>
>
>
> 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<http://www.natural-history-conservation.com>
>
>
> <image002.jpg>
>
>
>
>
> 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
>
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
>
>
>
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
> --
>
> <image004.png>
>
>
> Dirk Neumann
>
> Tel: 089 / 8107-111
> Fax: 089 / 8107-300
> neumann(a)snsb.de
>
> Postanschrift:
>
> Staatliche Naturwissenschaftliche Sammlungen Bayerns
> Zoologische Staatssammlung M?nchen
> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
> M?nchhausenstr. 21
> 81247 M?nchen
>
> Besuchen Sie unsere Sammlung:
> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>
> ---------
>
> 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)
>
> Visit our section at:
> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
>

--
Mare Nazaire, Ph.D.
Administrative Curator, Herbarium [RSA-POM]
California Botanic Garden
Research Assistant Professor, Claremont Graduate University
1500 North College Avenue
Claremont, California 91711
909.625.8767 ext. 268
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------------------------------

Message: 3
Date: Fri, 7 May 2021 11:43:32 -0400
From: John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
To: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>
Cc: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>, NHCOLL-new
    <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID:
    <CAF7GCDaQVa_-SWiu-GiNU0bYfgCUJOkrtRXoqAz9GQ_U6C8b-g at mail.gmail.com<mailto:CAF7GCDaQVa_-SWiu-GiNU0bYfgCUJOkrtRXoqAz9GQ_U6C8b-g at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

The reference for the book is:
Simmons, John E. 2014. *Fluid Preservation: A Comprehensive Reference*.
Rowman & Littlefield. It is available from Amazon.com, from the publisher,
and other sellers.

The book includes a discussion (pp 54-56) of botanical fluid preservation
(thanks to Ann Pinzl for generously sharing with me her as yet unpublished
research on this subject). Botanists have tended to use some strange
mixtures, trying to preserve color in their specimens (particularly in
flowers).

The use of beverage alcohol as a preservative has a long and fascinating
history. Although most beverage alcohol is below 70%, it usually does a
fairly good job of preservation (proof is approximately half the alcohol
concentration, so a 20 proof spirit is about 40% ETOH). I have used
beverage alcohol to preserve when nothing else was available, and have seen
a lot of specimens preserved in it. Rum was commonly used because it was
inexpensive (things such as brandy, which usually has a higher alcohol
content than rum, actually work better).

Over the history of fluid preservation, there have been many attempts to
improve the preservative properties of alcohol. In the days before we had
an easy means to check the concentration, it was common to use alcohol
after a second distillation, which usually meant around 60-65% (depending
on the source), so such things as arsenic, mercuric chloride, and other
chemicals were commonly added to "strengthen" it (they were really just
making it a more effective biocide, but usually screwing up the specimen in
the process).

--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 Fri, May 7, 2021 at 10:56 AM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:

> Thank you Simon and Dirk for your feedback on this.
>
> I had actually spoken with the botanist who originally prepared the
> specimens back in the 60's - he noted that they were preserved in 50% EtOH.
> He also noted that when he had traveled to other countries for field work
> and EtOH wasn't available he would use rum! So there could be some other
> residual components in these fluid preserved specimens!
>
> On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
> wrote:
>
>> Dear Mare,
>>
>> I have always fixed fresh plant material in Kew mix and then transferred
>> to Copenhagen mixture which is similar but minus the formalin.  As Dirk has
>> pointed out, the formulae (proportions) do vary slightly between
>> institutions, some prefer more glycerine in their mixes but which can make
>> the specimens rather translucent which is why others prefer a lower
>> concentration.  There is also the slight problem of osmotic pressure
>> differential and specimens floating until they equilibrate!
>> Make sure that the pH of the solutions is a near to 7.0 as possible.
>>
>> With all good wishes, Simon
>>
>> Simon Moore MIScT, RSci, FLS, ACR
>> Conservator of Natural Sciences and Cutlery Historian,
>>
>> www.natural-history-conservation.com<http://www.natural-history-conservation.com>
>>
>>
>>
>>
>> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>
>> 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 ?hlander <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 ?hlander
>>
>> 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>
>>
>>
>>
>>
>>
>>
>>
>>
>>
>> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
>> Skickat: den 7 maj 2021 08:36
>> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
>> ?mne: 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
>>
>>
>>
>>
>>
>>
>>
>> <image001.png>
>>
>>
>>
>> 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
>>
>>
>> <image002.jpg>
>>
>>
>>
>>
>> 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
>>
>>
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>>
>>
>>
>>
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>>
>> <image004.png>
>>
>>
>> Dirk Neumann
>>
>> Tel: 089 / 8107-111
>> Fax: 089 / 8107-300
>> neumann(a)snsb.de
>>
>> Postanschrift:
>>
>> Staatliche Naturwissenschaftliche Sammlungen Bayerns
>> Zoologische Staatssammlung M?nchen
>> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
>> M?nchhausenstr. 21
>> 81247 M?nchen
>>
>> Besuchen Sie unsere Sammlung:
>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>
>> ---------
>>
>> 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)
>>
>> Visit our section at:
>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>> Mare Nazaire, Ph.D.
>> Administrative Curator, Herbarium [RSA-POM]
>> California Botanic Garden
>> Research Assistant Professor, Claremont Graduate University
>> 1500 North College Avenue
>> Claremont, California 91711
>> 909.625.8767 ext. 268
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
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------------------------------

Message: 4
Date: Fri, 7 May 2021 09:02:29 -0700
From: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>
To: John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
Cc: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>, NHCOLL-new
    <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID:
    <CAGsrPoKnjZjcjyUNX656KfoyCryKSsN0FUtUW3Ak6frNgBqPzA at mail.gmail.com<mailto:CAGsrPoKnjZjcjyUNX656KfoyCryKSsN0FUtUW3Ak6frNgBqPzA at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Thank you for the citation and for all of this helpful information John!

On Fri, May 7, 2021 at 8:43 AM John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
wrote:

> The reference for the book is:
> Simmons, John E. 2014. *Fluid Preservation: A Comprehensive Reference*.
> Rowman & Littlefield. It is available from Amazon.com, from the publisher,
> and other sellers.
>
> The book includes a discussion (pp 54-56) of botanical fluid preservation
> (thanks to Ann Pinzl for generously sharing with me her as yet unpublished
> research on this subject). Botanists have tended to use some strange
> mixtures, trying to preserve color in their specimens (particularly in
> flowers).
>
> The use of beverage alcohol as a preservative has a long and fascinating
> history. Although most beverage alcohol is below 70%, it usually does a
> fairly good job of preservation (proof is approximately half the alcohol
> concentration, so a 20 proof spirit is about 40% ETOH). I have used
> beverage alcohol to preserve when nothing else was available, and have seen
> a lot of specimens preserved in it. Rum was commonly used because it was
> inexpensive (things such as brandy, which usually has a higher alcohol
> content than rum, actually work better).
>
> Over the history of fluid preservation, there have been many attempts to
> improve the preservative properties of alcohol. In the days before we had
> an easy means to check the concentration, it was common to use alcohol
> after a second distillation, which usually meant around 60-65% (depending
> on the source), so such things as arsenic, mercuric chloride, and other
> chemicals were commonly added to "strengthen" it (they were really just
> making it a more effective biocide, but usually screwing up the specimen in
> the process).
>
> --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 Fri, May 7, 2021 at 10:56 AM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>
>> Thank you Simon and Dirk for your feedback on this.
>>
>> I had actually spoken with the botanist who originally prepared the
>> specimens back in the 60's - he noted that they were preserved in 50% EtOH.
>> He also noted that when he had traveled to other countries for field work
>> and EtOH wasn't available he would use rum! So there could be some other
>> residual components in these fluid preserved specimens!
>>
>> On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
>> wrote:
>>
>>> Dear Mare,
>>>
>>> I have always fixed fresh plant material in Kew mix and then transferred
>>> to Copenhagen mixture which is similar but minus the formalin.  As Dirk has
>>> pointed out, the formulae (proportions) do vary slightly between
>>> institutions, some prefer more glycerine in their mixes but which can make
>>> the specimens rather translucent which is why others prefer a lower
>>> concentration.  There is also the slight problem of osmotic pressure
>>> differential and specimens floating until they equilibrate!
>>> Make sure that the pH of the solutions is a near to 7.0 as possible.
>>>
>>> With all good wishes, Simon
>>>
>>> Simon Moore MIScT, RSci, FLS, ACR
>>> Conservator of Natural Sciences and Cutlery Historian,
>>>
>>> www.natural-history-conservation.com
>>>
>>>
>>>
>>>
>>> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>>
>>> 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 ?hlander <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 ?hlander
>>>
>>> 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>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
>>> Skickat: den 7 maj 2021 08:36
>>> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
>>> ?mne: 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
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>> <image001.png>
>>>
>>>
>>>
>>> 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
>>>
>>>
>>> <image002.jpg>
>>>
>>>
>>>
>>>
>>> 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
>>>
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>>
>>>
>>>
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>> --
>>>
>>> <image004.png>
>>>
>>>
>>> Dirk Neumann
>>>
>>> Tel: 089 / 8107-111
>>> Fax: 089 / 8107-300
>>> neumann(a)snsb.de
>>>
>>> Postanschrift:
>>>
>>> Staatliche Naturwissenschaftliche Sammlungen Bayerns
>>> Zoologische Staatssammlung M?nchen
>>> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
>>> M?nchhausenstr. 21
>>> 81247 M?nchen
>>>
>>> Besuchen Sie unsere Sammlung:
>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>
>>> ---------
>>>
>>> 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)
>>>
>>> Visit our section at:
>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>> --
>>> Mare Nazaire, Ph.D.
>>> Administrative Curator, Herbarium [RSA-POM]
>>> California Botanic Garden
>>> Research Assistant Professor, Claremont Graduate University
>>> 1500 North College Avenue
>>> Claremont, California 91711
>>> 909.625.8767 ext. 268
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>>
>>
>> --
>> Mare Nazaire, Ph.D.
>> Administrative Curator, Herbarium [RSA-POM]
>> California Botanic Garden
>> Research Assistant Professor, Claremont Graduate University
>> 1500 North College Avenue
>> Claremont, California 91711
>> 909.625.8767 ext. 268
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>

--
Mare Nazaire, Ph.D.
Administrative Curator, Herbarium [RSA-POM]
California Botanic Garden
Research Assistant Professor, Claremont Graduate University
1500 North College Avenue
Claremont, California 91711
909.625.8767 ext. 268
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------------------------------

Message: 5
Date: Fri, 7 May 2021 12:21:02 -0400
From: John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
To: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>
Cc: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>, NHCOLL-new
    <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID:
    <CAF7GCDaA_bUkt+HwdB+XEj0OszL-5oPVFsdtVC=2NkzMfoneQA at mail.gmail.com<mailto:2NkzMfoneQA at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Oops, I made a mistake in my previous response. Thanks to Peter Rauch for
catching it:  Proof is actually about TWICE what the ETOH concentration is,
so a 40 proof liquor would be about 20% alcohol.

The concept of proof goes back to 1705 when some measure was needed to test
the concentration of alcohol. In England, 100 proof was defined as a
concentration of 11 parts alcohol and 10 parts water, which would permit
the ignition of gunpowder. The modern definition of proof is a mixture of
alcohol and water with a specific gravity of 0.91984, or about twice the
concentration of the alcohol.

--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 Fri, May 7, 2021 at 12:02 PM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
> Thank you for the citation and for all of this helpful information John!
>
> On Fri, May 7, 2021 at 8:43 AM John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
> wrote:
>
>> The reference for the book is:
>> Simmons, John E. 2014. *Fluid Preservation: A Comprehensive Reference*.
>> Rowman & Littlefield. It is available from Amazon.com, from the publisher,
>> and other sellers.
>>
>> The book includes a discussion (pp 54-56) of botanical fluid preservation
>> (thanks to Ann Pinzl for generously sharing with me her as yet unpublished
>> research on this subject). Botanists have tended to use some strange
>> mixtures, trying to preserve color in their specimens (particularly in
>> flowers).
>>
>> The use of beverage alcohol as a preservative has a long and fascinating
>> history. Although most beverage alcohol is below 70%, it usually does a
>> fairly good job of preservation (proof is approximately half the alcohol
>> concentration, so a 20 proof spirit is about 40% ETOH). I have used
>> beverage alcohol to preserve when nothing else was available, and have seen
>> a lot of specimens preserved in it. Rum was commonly used because it was
>> inexpensive (things such as brandy, which usually has a higher alcohol
>> content than rum, actually work better).
>>
>> Over the history of fluid preservation, there have been many attempts to
>> improve the preservative properties of alcohol. In the days before we had
>> an easy means to check the concentration, it was common to use alcohol
>> after a second distillation, which usually meant around 60-65% (depending
>> on the source), so such things as arsenic, mercuric chloride, and other
>> chemicals were commonly added to "strengthen" it (they were really just
>> making it a more effective biocide, but usually screwing up the specimen in
>> the process).
>>
>> --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 Fri, May 7, 2021 at 10:56 AM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>
>>> Thank you Simon and Dirk for your feedback on this.
>>>
>>> I had actually spoken with the botanist who originally prepared the
>>> specimens back in the 60's - he noted that they were preserved in 50% EtOH.
>>> He also noted that when he had traveled to other countries for field work
>>> and EtOH wasn't available he would use rum! So there could be some other
>>> residual components in these fluid preserved specimens!
>>>
>>> On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
>>> wrote:
>>>
>>>> Dear Mare,
>>>>
>>>> I have always fixed fresh plant material in Kew mix and then
>>>> transferred to Copenhagen mixture which is similar but minus the formalin.
>>>> As Dirk has pointed out, the formulae (proportions) do vary slightly
>>>> between institutions, some prefer more glycerine in their mixes but which
>>>> can make the specimens rather translucent which is why others prefer a
>>>> lower concentration.  There is also the slight problem of osmotic pressure
>>>> differential and specimens floating until they equilibrate!
>>>> Make sure that the pH of the solutions is a near to 7.0 as possible.
>>>>
>>>> With all good wishes, Simon
>>>>
>>>> Simon Moore MIScT, RSci, FLS, ACR
>>>> Conservator of Natural Sciences and Cutlery Historian,
>>>>
>>>> www.natural-history-conservation.com
>>>>
>>>>
>>>>
>>>>
>>>> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>>>
>>>> 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 ?hlander <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 ?hlander
>>>>
>>>> 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>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
>>>> Skickat: den 7 maj 2021 08:36
>>>> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
>>>> ?mne: 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
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>> <image001.png>
>>>>
>>>>
>>>>
>>>> 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
>>>>
>>>>
>>>> <image002.jpg>
>>>>
>>>>
>>>>
>>>>
>>>> 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
>>>>
>>>>
>>>> _______________________________________________
>>>> Nhcoll-l mailing list
>>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>>
>>>> _______________________________________________
>>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>>> Natural History Collections (SPNHC), an international society whose
>>>> mission is to improve the preservation, conservation and management of
>>>> natural history collections to ensure their continuing value to
>>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>>> Advertising on NH-COLL-L is inappropriate.
>>>> _______________________________________________
>>>> Nhcoll-l mailing list
>>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>>
>>>> _______________________________________________
>>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>>> Natural History Collections (SPNHC), an international society whose
>>>> mission is to improve the preservation, conservation and management of
>>>> natural history collections to ensure their continuing value to
>>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>>> Advertising on NH-COLL-L is inappropriate.
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>> _______________________________________________
>>>> Nhcoll-l mailing list
>>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>>
>>>> _______________________________________________
>>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>>> Natural History Collections (SPNHC), an international society whose
>>>> mission is to improve the preservation, conservation and management of
>>>> natural history collections to ensure their continuing value to
>>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>>> Advertising on NH-COLL-L is inappropriate.
>>>>
>>>>
>>>> --
>>>>
>>>> <image004.png>
>>>>
>>>>
>>>> Dirk Neumann
>>>>
>>>> Tel: 089 / 8107-111
>>>> Fax: 089 / 8107-300
>>>> neumann(a)snsb.de
>>>>
>>>> Postanschrift:
>>>>
>>>> Staatliche Naturwissenschaftliche Sammlungen Bayerns
>>>> Zoologische Staatssammlung M?nchen
>>>> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
>>>> M?nchhausenstr. 21
>>>> 81247 M?nchen
>>>>
>>>> Besuchen Sie unsere Sammlung:
>>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>>
>>>> ---------
>>>>
>>>> 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)
>>>>
>>>> Visit our section at:
>>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>>
>>>> _______________________________________________
>>>> Nhcoll-l mailing list
>>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>>
>>>> _______________________________________________
>>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>>> Natural History Collections (SPNHC), an international society whose
>>>> mission is to improve the preservation, conservation and management of
>>>> natural history collections to ensure their continuing value to
>>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>>> Advertising on NH-COLL-L is inappropriate.
>>>>
>>>>
>>>> --
>>>> Mare Nazaire, Ph.D.
>>>> Administrative Curator, Herbarium [RSA-POM]
>>>> California Botanic Garden
>>>> Research Assistant Professor, Claremont Graduate University
>>>> 1500 North College Avenue
>>>> Claremont, California 91711
>>>> 909.625.8767 ext. 268
>>>> _______________________________________________
>>>> Nhcoll-l mailing list
>>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>>
>>>> _______________________________________________
>>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>>> Natural History Collections (SPNHC), an international society whose
>>>> mission is to improve the preservation, conservation and management of
>>>> natural history collections to ensure their continuing value to
>>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>>> Advertising on NH-COLL-L is inappropriate.
>>>>
>>>>
>>>>
>>>
>>> --
>>> Mare Nazaire, Ph.D.
>>> Administrative Curator, Herbarium [RSA-POM]
>>> California Botanic Garden
>>> Research Assistant Professor, Claremont Graduate University
>>> 1500 North College Avenue
>>> Claremont, California 91711
>>> 909.625.8767 ext. 268
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268
>
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------------------------------

Message: 6
Date: Fri, 7 May 2021 17:45:57 +0100
From: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
To: John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
Cc: Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>>, NHCOLL-new
    <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID: <A4B2DD3D-DE19-4C56-9A22-0567E0D1956B at btinternet.com<mailto:A4B2DD3D-DE19-4C56-9A22-0567E0D1956B at btinternet.com>>
Content-Type: text/plain;    charset=utf-8

That?s true John,

You have to understand that anything weights and measures-wise that emanated from England before about 1900 was bound to be complicated!

With all good wishes, Simon

> On 7 May 2021, at 17:21, John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>> wrote:
>
> Oops, I made a mistake in my previous response. Thanks to Peter Rauch for catching it:  Proof is actually about TWICE what the ETOH concentration is, so a 40 proof liquor would be about 20% alcohol.
>
> The concept of proof goes back to 1705 when some measure was needed to test the concentration of alcohol. In England, 100 proof was defined as a concentration of 11 parts alcohol and 10 parts water, which would permit the ignition of gunpowder. The modern definition of proof is a mixture of alcohol and water with a specific gravity of 0.91984, or about twice the concentration of the alcohol.
>
> --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 Fri, May 7, 2021 at 12:02 PM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
> Thank you for the citation and for all of this helpful information John!
>
> On Fri, May 7, 2021 at 8:43 AM John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>> wrote:
> The reference for the book is:
> Simmons, John E. 2014. Fluid Preservation: A Comprehensive Reference. Rowman & Littlefield. It is available from Amazon.com, from the publisher, and other sellers.
>
> The book includes a discussion (pp 54-56) of botanical fluid preservation (thanks to Ann Pinzl for generously sharing with me her as yet unpublished research on this subject). Botanists have tended to use some strange mixtures, trying to preserve color in their specimens (particularly in flowers).
>
> The use of beverage alcohol as a preservative has a long and fascinating history. Although most beverage alcohol is below 70%, it usually does a fairly good job of preservation (proof is approximately half the alcohol concentration, so a 20 proof spirit is about 40% ETOH). I have used beverage alcohol to preserve when nothing else was available, and have seen a lot of specimens preserved in it. Rum was commonly used because it was inexpensive (things such as brandy, which usually has a higher alcohol content than rum, actually work better).
>
> Over the history of fluid preservation, there have been many attempts to improve the preservative properties of alcohol. In the days before we had an easy means to check the concentration, it was common to use alcohol after a second distillation, which usually meant around 60-65% (depending on the source), so such things as arsenic, mercuric chloride, and other chemicals were commonly added to "strengthen" it (they were really just making it a more effective biocide, but usually screwing up the specimen in the process).
>
> --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 Fri, May 7, 2021 at 10:56 AM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
> Thank you Simon and Dirk for your feedback on this.
>
> I had actually spoken with the botanist who originally prepared the specimens back in the 60's - he noted that they were preserved in 50% EtOH. He also noted that when he had traveled to other countries for field work and EtOH wasn't available he would use rum! So there could be some other residual components in these fluid preserved specimens!
>
> On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>> wrote:
> Dear Mare,
>
> I have always fixed fresh plant material in Kew mix and then transferred to Copenhagen mixture which is similar but minus the formalin.  As Dirk has pointed out, the formulae (proportions) do vary slightly between institutions, some prefer more glycerine in their mixes but which can make the specimens rather translucent which is why others prefer a lower concentration.  There is also the slight problem of osmotic pressure differential and specimens floating until they equilibrate!
> Make sure that the pH of the solutions is a near to 7.0 as possible.
>
> With all good wishes, Simon
>
> Simon Moore MIScT, RSci, FLS, ACR
> Conservator of Natural Sciences and Cutlery Historian,
>
> www.natural-history-conservation.com
>
>
> <PastedGraphic-2.tiff><MA logo.jpg>
>
>> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>
>> 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 ?hlander <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 ?hlander
>>
>> 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>
>>
>>
>>
>>
>>
>>
>>
>>
>>
>> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
>> Skickat: den 7 maj 2021 08:36
>> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
>> ?mne: 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
>>
>>
>>
>>
>>
>>
>>
>> <image001.png>
>>
>>
>>
>> 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
>>
>>
>> <image002.jpg>
>>
>>
>>
>>
>> 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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>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
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>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>>
>>
>>
>>
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>>
>> <image004.png>
>>
>>
>> Dirk Neumann
>>
>> Tel: 089 / 8107-111
>> Fax: 089 / 8107-300
>> neumann(a)snsb.de
>>
>> Postanschrift:
>>
>> Staatliche Naturwissenschaftliche Sammlungen Bayerns
>> Zoologische Staatssammlung M?nchen
>> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
>> M?nchhausenstr. 21
>> 81247 M?nchen
>>
>> Besuchen Sie unsere Sammlung:
>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>
>> ---------
>>
>> 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)
>>
>> Visit our section at:
>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>> Mare Nazaire, Ph.D.
>> Administrative Curator, Herbarium [RSA-POM]
>> California Botanic Garden
>> Research Assistant Professor, Claremont Graduate University
>> 1500 North College Avenue
>> Claremont, California 91711
>> 909.625.8767 ext. 268
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>
>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
>
> --
> Mare Nazaire, Ph.D.
> Administrative Curator, Herbarium [RSA-POM]
> California Botanic Garden
> Research Assistant Professor, Claremont Graduate University
> 1500 North College Avenue
> Claremont, California 91711
> 909.625.8767 ext. 268

------------------------------

Message: 7
Date: Fri, 7 May 2021 19:18:57 +0200
From: Dirk Neumann <neumann at snsb.de<mailto:neumann at snsb.de>>
To: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: Re: [Nhcoll-l] Alcohol concentration for terrestrial
    vertebrates
Message-ID: <df425eb2-b921-2131-2336-0258a87722a0 at snsb.de<mailto:df425eb2-b921-2131-2336-0258a87722a0 at snsb.de>>
Content-Type: text/plain; charset="utf-8"; Format="flowed"

Maybe things started to go wrong after the deep trauma Isaac got when he
was hit by this apple? Gradually, and after a rich experience of
applying all sorts of interesting calculations, conversions and
transformations, the English are surfacing on the metric world.

;-)

With best wishes
Dirk

Am 07.05.2021 um 18:45 schrieb Simon Moore:
> That?s true John,
>
> You have to understand that anything weights and measures-wise that emanated from England before about 1900 was bound to be complicated!
>
> With all good wishes, Simon
>
>
>
>> On 7 May 2021, at 17:21, John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>> wrote:
>>
>> Oops, I made a mistake in my previous response. Thanks to Peter Rauch for catching it:  Proof is actually about TWICE what the ETOH concentration is, so a 40 proof liquor would be about 20% alcohol.
>>
>> The concept of proof goes back to 1705 when some measure was needed to test the concentration of alcohol. In England, 100 proof was defined as a concentration of 11 parts alcohol and 10 parts water, which would permit the ignition of gunpowder. The modern definition of proof is a mixture of alcohol and water with a specific gravity of 0.91984, or about twice the concentration of the alcohol.
>>
>> --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 Fri, May 7, 2021 at 12:02 PM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>> Thank you for the citation and for all of this helpful information John!
>>
>> On Fri, May 7, 2021 at 8:43 AM John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>> wrote:
>> The reference for the book is:
>> Simmons, John E. 2014. Fluid Preservation: A Comprehensive Reference. Rowman & Littlefield. It is available from Amazon.com, from the publisher, and other sellers.
>>
>> The book includes a discussion (pp 54-56) of botanical fluid preservation (thanks to Ann Pinzl for generously sharing with me her as yet unpublished research on this subject). Botanists have tended to use some strange mixtures, trying to preserve color in their specimens (particularly in flowers).
>>
>> The use of beverage alcohol as a preservative has a long and fascinating history. Although most beverage alcohol is below 70%, it usually does a fairly good job of preservation (proof is approximately half the alcohol concentration, so a 20 proof spirit is about 40% ETOH). I have used beverage alcohol to preserve when nothing else was available, and have seen a lot of specimens preserved in it. Rum was commonly used because it was inexpensive (things such as brandy, which usually has a higher alcohol content than rum, actually work better).
>>
>> Over the history of fluid preservation, there have been many attempts to improve the preservative properties of alcohol. In the days before we had an easy means to check the concentration, it was common to use alcohol after a second distillation, which usually meant around 60-65% (depending on the source), so such things as arsenic, mercuric chloride, and other chemicals were commonly added to "strengthen" it (they were really just making it a more effective biocide, but usually screwing up the specimen in the process).
>>
>> --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 Fri, May 7, 2021 at 10:56 AM Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>> Thank you Simon and Dirk for your feedback on this.
>>
>> I had actually spoken with the botanist who originally prepared the specimens back in the 60's - he noted that they were preserved in 50% EtOH. He also noted that when he had traveled to other countries for field work and EtOH wasn't available he would use rum! So there could be some other residual components in these fluid preserved specimens!
>>
>> On Fri, May 7, 2021 at 7:18 AM Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>> wrote:
>> Dear Mare,
>>
>> I have always fixed fresh plant material in Kew mix and then transferred to Copenhagen mixture which is similar but minus the formalin.  As Dirk has pointed out, the formulae (proportions) do vary slightly between institutions, some prefer more glycerine in their mixes but which can make the specimens rather translucent which is why others prefer a lower concentration.  There is also the slight problem of osmotic pressure differential and specimens floating until they equilibrate!
>> Make sure that the pH of the solutions is a near to 7.0 as possible.
>>
>> With all good wishes, Simon
>>
>> Simon Moore MIScT, RSci, FLS, ACR
>> Conservator of Natural Sciences and Cutlery Historian,
>>
>> www.natural-history-conservation.com
>>
>>
>> <PastedGraphic-2.tiff><MA logo.jpg>
>>
>>> On 7 May 2021, at 13:53, Mare Nazaire <mnazaire at calbg.org<mailto:mnazaire at calbg.org>> wrote:
>>>
>>> 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 ?hlander <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 ?hlander
>>>
>>> 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>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>> Fr?n: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> F?r Dirk Neumann
>>> Skickat: den 7 maj 2021 08:36
>>> Till: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
>>> ?mne: 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
>>>
>>>
>>>
>>>
>>>
>>>
>>>
>>> <image001.png>
>>>
>>>
>>>
>>> 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
>>>
>>>
>>> <image002.jpg>
>>>
>>>
>>>
>>>
>>> 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
>>>
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>>
>>>
>>>
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>> --
>>>
>>> <image004.png>
>>>
>>>
>>> Dirk Neumann
>>>
>>> Tel: 089 / 8107-111
>>> Fax: 089 / 8107-300
>>> neumann(a)snsb.de
>>>
>>> Postanschrift:
>>>
>>> Staatliche Naturwissenschaftliche Sammlungen Bayerns
>>> Zoologische Staatssammlung M?nchen
>>> Dirk Neumann, Sektion Ichthyologie / DNA-Storage
>>> M?nchhausenstr. 21
>>> 81247 M?nchen
>>>
>>> Besuchen Sie unsere Sammlung:
>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>
>>> ---------
>>>
>>> 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)
>>>
>>> Visit our section at:
>>> http://www.zsm.mwn.de/sektion/ichthyologie-home/
>>>
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>>
>>>
>>> --
>>> Mare Nazaire, Ph.D.
>>> Administrative Curator, Herbarium [RSA-POM]
>>> California Botanic Garden
>>> Research Assistant Professor, Claremont Graduate University
>>> 1500 North College Avenue
>>> Claremont, California 91711
>>> 909.625.8767 ext. 268
>>> _______________________________________________
>>> Nhcoll-l mailing list
>>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>>
>>> _______________________________________________
>>> NHCOLL-L is brought to you by the Society for the Preservation of
>>> Natural History Collections (SPNHC), an international society whose
>>> mission is to improve the preservation, conservation and management of
>>> natural history collections to ensure their continuing value to
>>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>> Mare Nazaire, Ph.D.
>> Administrative Curator, Herbarium [RSA-POM]
>> California Botanic Garden
>> Research Assistant Professor, Claremont Graduate University
>> 1500 North College Avenue
>> Claremont, California 91711
>> 909.625.8767 ext. 268
>> _______________________________________________
>> Nhcoll-l mailing list
>> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
>> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>>
>> _______________________________________________
>> NHCOLL-L is brought to you by the Society for the Preservation of
>> Natural History Collections (SPNHC), an international society whose
>> mission is to improve the preservation, conservation and management of
>> natural history collections to ensure their continuing value to
>> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
>> Advertising on NH-COLL-L is inappropriate.
>>
>>
>> --
>> Mare Nazaire, Ph.D.
>> Administrative Curator, Herbarium [RSA-POM]
>> California Botanic Garden
>> Research Assistant Professor, Claremont Graduate University
>> 1500 North College Avenue
>> Claremont, California 91711
>> 909.625.8767 ext. 268
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.

--

Dirk Neumann

Tel: 089 / 8107-111
Fax: 089 / 8107-300
neumann(a)snsb.de

Postanschrift:

Staatliche Naturwissenschaftliche Sammlungen Bayerns
Zoologische Staatssammlung M?nchen
Dirk Neumann, Sektion Ichthyologie / DNA-Storage
M?nchhausenstr. 21
81247 M?nchen

Besuchen Sie unsere Sammlung:
http://www.zsm.mwn.de/sektion/ichthyologie-home/

---------

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)

Visit our section at:
http://www.zsm.mwn.de/sektion/ichthyologie-home/

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Message: 8
Date: Fri, 7 May 2021 15:12:56 -0400
From: Jyotsna Pandey <jpandey at aibs.org<mailto:jpandey at aibs.org>>
To: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: [Nhcoll-l] AIBS Writing for Impact and Influence Online
    Course: Register Now
Message-ID:
    <CAOuC=0HrCA2ATBS5JsJ7rWR879rKEdDzeVPxqszFYacTDc6rpg at mail.gmail.com<mailto:0HrCA2ATBS5JsJ7rWR879rKEdDzeVPxqszFYacTDc6rpg at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Dear Colleagues,

There is a growing recognition of the importance of providing scientists,
particularly graduate students and post-doctoral fellows, with professional
development training that will expand their career opportunities and
potential for professional success.  The American Institute of Biological
Sciences (AIBS) is pleased to announce a professional development program
that we have developed to help scientists strengthen their written
communication
skills.

This is an important professional development training opportunity.  I hope
you will consider sharing this opportunity with your students, staff, and
colleagues.

Below are more specific details about this online course, including
registration information.

*Writing for Impact and Influence: An AIBS Professional Development Program*

*It is perfectly okay to write garbage?as long as you edit brilliantly. *
-C. J. Cherryh

The American Institute of Biological Sciences (AIBS) has heard a common
refrain from faculty, scientists, government and private sector executives,
and everyone in between: Scientists are increasingly responsible for public
engagement and business writing, yet they are rarely given the tools they
need to succeed.

AIBS is responding by re-offering our professional development program
designed to help scientists, including graduate students, hone their
written communication skills to increase the impact and influence of their
message. This course complements AIBS?s highly successful Communications
Boot Camp for Scientists, which focuses on oral communication.

Writing for Impact and Influence provides practical instruction and
hands-on exercises that will improve the participant?s general writing
proficiency. The program will provide participants with the skills and
tools needed to compose scientific press releases, blog posts, memoranda,
and more, with a focus on the reader experience. Each product-oriented
session will have an assignment (deadlines are flexible), with feedback
from the instructor. The course is interactive, and participants are
encouraged to ask questions and exchange ideas with the instructor and
other participants. Each session is also recorded and shared with all
participants to accommodate scheduling conflicts.

*Who Should Take the Course?*

  - Individuals interested in furthering their professional development by
  augmenting their writing skills.
  - Graduate students and early-career professionals interested in
  increasing their marketability to employers.
  - Individuals interested in more effectively informing and influencing
  segments of the public, supervisors, policymakers, reporters,
  organizational leaders, and others.

*Sample Topics*

  - Press releases and writing for the media
  - Blogging and social media campaigns
  - Writing for professional audiences
  - One-pagers and writing for stakeholders
  - Action/decision memoranda
  - Synthesis

*Course Structure*
The course consists of six 90-minute online modules conducted live and
subsequently archived online for participant review. Modules are spaced at
weekly intervals to allow time for assignment completion. Live attendance
is recommended but not required, and the instructor can be contacted by
email at any time during the course.

*Assignments*
A writing assignment will be given in each of the first five courses.
Students will receive timely feedback on their assignments.

*Schedule*
The course will begin on Thursday, 1 July 2021. The subsequent course
sessions will be held weekly on Thursdays, through 5 August. All live
courses will begin at 2:00 p.m. Eastern time. Recorded programs will be
available to participants after the live session.

*Registration*
Space is limited and the course will be filled on a first-come,
first-served basis. Registration is required. To register for the course,
go to http://io.aibs.org/writing

For questions regarding the course please contact James Verdier at
jverdier at aibs.org.<mailto:jverdier at aibs.org.>

Sincerely,
Jyotsna
___________________
Jyotsna Pandey, Ph.D.
Director of Public Policy, American Institute of Biological Sciences (AIBS)
950 Herndon Parkway Suite 450
Herndon, VA 20170
Phone: 202-628-1500 x 225
AIBS website: www.aibs.org
Follow AIBS on Twitter! @AIBSbiology <https://twitter.com/aibsbiology>

--
This message is confidential and should only be read by its intended
recipients.?
If you have received it in error, please notify the sender and
delete all copies.
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Message: 9
Date: Fri, 7 May 2021 15:36:40 -0500
From: Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>>
To: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: [Nhcoll-l] Alcohol concentrations for preserving flowers
Message-ID:
    <CA+Wdk7m_djb2WYgHRxmLnsaGGcfXv81GM6f+G0jOwFy9xe=g1g at mail.gmail.com<mailto:g1g at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Hi there!
Is there a recommended ETOH concentration for preserving flowers for later
dissection? In a pinch, I've used cheap vodka (about 40%) but I'm not sure
I want to bring that into a classroom.  Any suggestions?

--
Gretchen Meier (*she/her/hers*)
Olga Lakela Herbarium (DUL)
University of Minnesota Duluth
gameier at d.umn.edu<mailto:gameier at d.umn.edu>
218.726.6542

*Naturum expellas furca, tamen usque recurret - Homer *
'you may drive nature out with a pitchfork, but she will come hurrying back'
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Message: 10
Date: Fri, 7 May 2021 21:07:11 +0000
From: "Bentley, Andrew Charles" <abentley at ku.edu<mailto:abentley at ku.edu>>
To: "nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>" <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: [Nhcoll-l] SPNHC opening mixer
Message-ID: <2C40737A-5902-4FFF-84CD-91E8912F1199 at ku.edu<mailto:2C40737A-5902-4FFF-84CD-91E8912F1199 at ku.edu>>
Content-Type: text/plain; charset="utf-8"

Hi all

There appears to be an issue with the link to the SPNHC mixer on the AIC website.  If you are trying to join us go here:  https://www.kumospace.com/spnhc2021

Andy

    A  :                A  :              A  :
}<(((_?>.,.,.,.}<(((_?>.,.,.,.}<)))_?>
    V                  V                  V
Andy Bentley
Ichthyology Collection Manager
University of Kansas
Biodiversity Institute
Dyche Hall
1345 Jayhawk Boulevard
Lawrence, KS, 66045-7561
USA

Tel: (785) 864-3863
Fax: (785) 864-5335
Email: abentley at ku.edu<mailto:abentley at ku.edu><mailto:abentley at ku.edu<mailto:abentley at ku.edu>>
http://ichthyology.biodiversity.ku.edu<http://ichthyology.biodiversity.ku.edu/>

    A  :                A  :                A  :
}<(((_?>.,.,.,.}<(((_?>.,.,.,.}<)))_?>
    V                  V                  V

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Message: 11
Date: Sat, 8 May 2021 12:27:21 +0100
From: Simon Moore <couteaufin at btinternet.com<mailto:couteaufin at btinternet.com>>
To: Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>>
Cc: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: Re: [Nhcoll-l] Alcohol concentrations for preserving flowers
Message-ID: <D26CCE6C-9073-4815-9714-B1E69BC50B38 at btinternet.com<mailto:D26CCE6C-9073-4815-9714-B1E69BC50B38 at btinternet.com>>
Content-Type: text/plain; charset="us-ascii"

Hi Gretchen
This is a tricky one as preservation of fugitive pigments that flowers contain is a bit of a biochemical mixture.  Someone may have experience of this but the slightly oily texture of vodka might be good as a short term preservative. Adding some Bitrex, denatonium benzoate, should discourage anyone sampling! Or use tubes with tight closures?

With all good wishes, Simon
Simon Moore MiSct, FLS, RRScI, ACR,
Conservator of natural sciences
Sent from my iPad

> On 7 May 2021, at 21:36, Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>> wrote:
>
> Hi there!
> Is there a recommended ETOH concentration for preserving flowers for later dissection? In a pinch, I've used cheap vodka (about 40%) but I'm not sure I want to bring that into a classroom.  Any suggestions?
>
> --
> Gretchen Meier (she/her/hers)
> Olga Lakela Herbarium (DUL)
> University of Minnesota Duluth
> gameier at d.umn.edu<mailto:gameier at d.umn.edu>
> 218.726.6542
>
> Naturum expellas furca, tamen usque recurret - Homer
> 'you may drive nature out with a pitchfork, but she will come hurrying back'
>
>
>
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
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Message: 12
Date: Mon, 10 May 2021 16:21:18 +0000
From: Nikolaj Scharff <nscharff at snm.ku.dk<mailto:nscharff at snm.ku.dk>>
To: "nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>" <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: [Nhcoll-l] Head of Conservation Unit at the Natural History
    Museum of Denmark (NHMD)
Message-ID: <955e7537e7fc49ca8e5030a8252397bb at snm.ku.dk<mailto:955e7537e7fc49ca8e5030a8252397bb at snm.ku.dk>>
Content-Type: text/plain; charset="us-ascii"

The Natural History Museum of Denmark (NHMD) is seeking an experienced natural history conservator to establish and manage a Conservation Unit which is to be established at the NHMD.

In the coming years, the primary focus of the Conservation Unit will be to conserve, prepare and install objects for the new permanent galleries which are currently being developed in connection with the build of a new national Natural History Museum in the midst of Copenhagen. Currently, seven exhibition core groups are developing the content of the new permanent galleries, which are planned to open to the public in October 2024 (https://nyt.snm.ku.dk/english/).

An exciting opportunity to be part of the new museum project. Check the job announcement below!

https://candidate.hr-manager.net/ApplicationInit.aspx/?cid=1307&departmentId=19220&ProjectId=154054&MediaId=5&SkipAdvertisement=false

Best wishes
Nikolaj Scharff

----------------------------------------------------------
Nikolaj Scharff, PhD
Deputy Museum Director
Professor, Head of Collections & Research

Natural History Museum of Denmark
Zoological Museum, University of Copenhagen
Universitetsparken 15, DK-2100 Copenhagen
DENMARK

E-mail: nscharff at snm.ku.dk<mailto:nscharff at snm.ku.dk><mailto:nscharff at snm.ku.dk<mailto:nscharff at snm.ku.dk>>
Webpage: http://snm.ku.dk/people/nscharff
----------------------------------------------------------

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Message: 13
Date: Mon, 10 May 2021 14:10:51 -0400
From: John E Simmons <simmons.johne at gmail.com<mailto:simmons.johne at gmail.com>>
To: Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>>
Cc: NHCOLL-new <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] Alcohol concentrations for preserving flowers
Message-ID:
    <CAF7GCDbrLDSpFEYuRA5Hi1bg5J3oPOppJj=29_ijOokGq84sUw at mail.gmail.com<mailto:29_ijOokGq84sUw at mail.gmail.com>>
Content-Type: text/plain; charset="utf-8"

Gretchen,

I was hoping that a botanist might respond on the list with a
recommendation for you (botany is not my specialty). I can tell you that
the original Kew mixture was 53 percent industrial methylated spirits (IMS,
which is ethyl alcohol containing 9 percent water and 2?4 percent
methanol), 37 percent water, 5 percent formalin (?dilute formaldehyde?),
and 5 percent glycerol (Forman and Bridson 1989).

A note in the second edition of *The Herbarium Handbook* (1992) stated that
due to controls on hazardous chemicals, the original formula at Kew had
been withdrawn and replaced by a mixture of ?70% alcohol (ethanol, or
ethanol + trace of methanol), 29% water and 1% glycerol. If using 90%
strength Industrial Methylated Spirit (IMS) the mixture is 78% IMS, 21%
water and 1% glycerol. With different strengths of IMS the proportions must
be adjusted accordingly? (Forman and Bridson 1989, 210).

In the third edition (Bridson and Forman 1998) the Kew mixture recipe was
ten parts IMS, one part formalin, one part glycerol, and eight parts water.

The Kew mixture was supposed to help preserve the color in plants. If your
concern is just to preserve flowers sufficiently for them to be dissected,
you might try 70% ETOH, with no other additions. If the specimens are not
intended to be saved as museum specimens, you could use denatured 70% ETOH,
which is considerably less expensive.

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 Fri, May 7, 2021 at 4:37 PM Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>> wrote:

> Hi there!
> Is there a recommended ETOH concentration for preserving flowers for later
> dissection? In a pinch, I've used cheap vodka (about 40%) but I'm not sure
> I want to bring that into a classroom.  Any suggestions?
>
> --
> Gretchen Meier (*she/her/hers*)
> Olga Lakela Herbarium (DUL)
> University of Minnesota Duluth
> gameier at d.umn.edu<mailto:gameier at d.umn.edu>
> 218.726.6542
>
> *Naturum expellas furca, tamen usque recurret - Homer *
> 'you may drive nature out with a pitchfork, but she will come hurrying
> back'
>
>
>
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.
>
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Message: 14
Date: Mon, 10 May 2021 22:35:01 +0200
From: Dirk Neumann <neumann at snsb.de<mailto:neumann at snsb.de>>
To: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: Re: [Nhcoll-l] Alcohol concentrations for preserving flowers
Message-ID: <4c0225cc-fd76-f642-f9d6-5e3ee84225a8 at snsb.de<mailto:4c0225cc-fd76-f642-f9d6-5e3ee84225a8 at snsb.de>>
Content-Type: text/plain; charset="utf-8"; Format="flowed"

... it might be worth adding that some denaturants as some ketones are
also used as component in chromatography and extract pigments quite well
(not all, put some). Natural colours can be caused by quite different
effects and sources, e.g., pigments, unsaturated [metal] ions as key
element in chelating agents, optical refraction on crystals embedded in
tissues, etc..

Often, the effect of visual colour we see in the specimens we collect is
caused by electron excitation in biomolecules, which cause changes in
the bonding or chemistry of respective molecules. Most of our fixatives
and storage fluids tend to be highly polar, and through polarisation or
depolarisation of respective biomolecules, colours usually fade - cf.
colour reaction in many indicators we use.

There are few natural pigments that are stable, but admittedly few. And
I would always be careful with bright coloured, historic teaching
specimens prepared around 1900 ... As old bold bright paints, the
ingredient are rarely healthy (and usually containing heavy metals such
as cadmium, chrome, lead, etc.)

With best wishes
Dirk

Am 10.05.2021 um 20:10 schrieb John E Simmons:
> Gretchen,
>
> I was hoping that a botanist might respond on the list with a
> recommendation for you (botany is not my specialty). I can tell you
> that the original Kew mixture was 53 percent industrial methylated
> spirits (IMS, which is ethyl alcohol containing 9 percent water and
> 2?4 percent methanol), 37 percent water, 5 percent formalin (?dilute
> formaldehyde?), and 5 percent glycerol (Forman and Bridson 1989).
>
> A note in the second edition of /The Herbarium Handbook/ (1992) stated
> that due to controls on hazardous chemicals, the original formula at
> Kew had been withdrawn and replaced by a mixture of ?70% alcohol
> (ethanol, or ethanol + trace of methanol), 29% water and 1% glycerol.
> If using 90% strength Industrial Methylated Spirit (IMS) the mixture
> is 78% IMS, 21% water and 1% glycerol. With different strengths of IMS
> the proportions must be adjusted accordingly? (Forman and Bridson
> 1989, 210).
>
> In the third edition (Bridson and Forman 1998) the Kew mixture recipe
> was ten parts IMS, one part formalin, one part glycerol, and eight
> parts water.
>
>
> The Kew mixture was supposed to help preserve the color in plants. If
> your concern is just to preserve flowers sufficiently for them to be
> dissected, you might try 70% ETOH, with no other additions. If the
> specimens are not intended to be saved as museum specimens, you could
> use denatured 70% ETOH, which is considerably less expensive.
>
>
> 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 Fri, May 7, 2021 at 4:37 PM Gretchen Meier <gameier at d.umn.edu<mailto:gameier at d.umn.edu>
> <mailto:gameier at d.umn.edu<mailto:gameier at d.umn.edu>>> wrote:
>
>    Hi there!
>    Is there a recommended ETOH concentration for preserving flowers
>    for later dissection? In a pinch, I've used cheap vodka (about
>    40%) but I'm not sure I want to bring that into a classroom.? Any
>    suggestions?
>
>    --
>    Gretchen Meier (/she/her/hers/)
>    Olga Lakela Herbarium (DUL)
>    University of Minnesota Duluth
>    gameier at d.umn.edu<mailto:gameier at d.umn.edu> <mailto:gameier at d.umn.edu<mailto:gameier at d.umn.edu>>
>    218.726.6542
>
>    /Naturum expellas furca, tamen usque recurret - Homer /
>    'you may drive nature out with a pitchfork, but she will come
>    hurrying back'
>
>
>
>    _______________________________________________
>    Nhcoll-l mailing list
>    Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu> <mailto:Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>>
>    https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>    <https://mailman.yale.edu/mailman/listinfo/nhcoll-l>
>
>    _______________________________________________
>    NHCOLL-L is brought to you by the Society for the Preservation of
>    Natural History Collections (SPNHC), an international society whose
>    mission is to improve the preservation, conservation and management of
>    natural history collections to ensure their continuing value to
>    society. See http://www.spnhc.org <http://www.spnhc.org%20> <http://www.spnhc.org> for
>    membership information.
>    Advertising on NH-COLL-L is inappropriate.
>
>
> _______________________________________________
> Nhcoll-l mailing list
> Nhcoll-l at mailman.yale.edu<mailto:Nhcoll-l at mailman.yale.edu>
> https://mailman.yale.edu/mailman/listinfo/nhcoll-l
>
> _______________________________________________
> NHCOLL-L is brought to you by the Society for the Preservation of
> Natural History Collections (SPNHC), an international society whose
> mission is to improve the preservation, conservation and management of
> natural history collections to ensure their continuing value to
> society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
> Advertising on NH-COLL-L is inappropriate.

--

Dirk Neumann

Tel: 089 / 8107-111
Fax: 089 / 8107-300
neumann(a)snsb.de

Postanschrift:

Staatliche Naturwissenschaftliche Sammlungen Bayerns
Zoologische Staatssammlung M?nchen
Dirk Neumann, Sektion Ichthyologie / DNA-Storage
M?nchhausenstr. 21
81247 M?nchen

Besuchen Sie unsere Sammlung:
http://www.zsm.mwn.de/sektion/ichthyologie-home/

---------

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)

Visit our section at:
http://www.zsm.mwn.de/sektion/ichthyologie-home/

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Message: 15
Date: Tue, 11 May 2021 19:27:16 -0400 (EDT)
From: "membership at spnhc.org<mailto:membership at spnhc.org>" <membership at spnhc.org<mailto:membership at spnhc.org>>
To: nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>
Subject: [Nhcoll-l] SPNHC Education DemoCamp 2021
Message-ID: <1620775636.256320094 at apps.rackspace.com<mailto:1620775636.256320094 at apps.rackspace.com>>
Content-Type: text/plain; charset="utf-8"

The Society for the Preservation of Natural History Collections Education Sessional Committee is hosting a virtual [ Natural History Collection Education DemoCamp
]( https://spnhc.org/education-democamp/ <https://spnhc.org/education-democamp/%20> )The goal is to share, discover, and discuss educational materials that have a framework in natural history. This is building upon our previous iterations of the 'education share fair' that were hosted at the annual meeting.

Where: Zoom, of course!

When: 28th and 29th of June, the amount of time blocks will be dependent on the number of presenters.

Cost: Free!

[ Presenter registration ]( https://docs.google.com/forms/u/1/d/e/1FAIpQLSc-wvV0pxDutboRlywLETIICxEUOOFkJOAd7eAlms5lIdtSqQ/viewform <https://docs.google.com/forms/u/1/d/e/1FAIpQLSc-wvV0pxDutboRlywLETIICxEUOOFkJOAd7eAlms5lIdtSqQ/viewform%20> ) is open until June 6th and[  general attendee registration ]( https://us02web.zoom.us/meeting/register/tZItcuqtqjMsG9GhIqNsWpIaNf7ilXh-gVeF <https://us02web.zoom.us/meeting/register/tZItcuqtqjMsG9GhIqNsWpIaNf7ilXh-gVeF%20> ) will be open up until the event.Please see the [ website  ]( https://spnhc.org/education-democamp/ <https://spnhc.org/education-democamp/%20> )for additional details and reach out to us with any questions: [ educationdemocamp at gmail.com<mailto:educationdemocamp at gmail.com> ]( mailto:educationdemocamp at gmail.com<mailto:educationdemocamp at gmail.com> )

Anna Monfils & Molly Philips, Committee Co-Chairs
Jen Bauer & Liz Leith, Committee Co-Secretary
Julie Robinson, Jessa Watters, & Kari Harris, Committee Members
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Message: 16
Date: Wed, 12 May 2021 04:35:10 +0000
From: "Haff, Tonya (NCMI, Crace)" <Tonya.Haff at csiro.au<mailto:Tonya.Haff at csiro.au>>
To: "nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>" <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: [Nhcoll-l] freezing blown eggs
Message-ID:
    <ME2PR01MB2676989C0A01F44E615DE9F6EE529 at ME2PR01MB2676.ausprd01.prod.outlook.com<mailto:ME2PR01MB2676989C0A01F44E615DE9F6EE529 at ME2PR01MB2676.ausprd01.prod.outlook.com>>

Content-Type: text/plain; charset="us-ascii"

Hello all,

I am wondering if any of you have thoughts on freezing egg collections for pest control? I imagine that it is probably ok, but then I wonder if there could be problems to the shell structure. Note I realise that there is not much for museum pests to eat in a blown egg, but there are lots of places for them to hide in and around the their housing. As usual, any thoughts or feedback would be very appreciated!

Cheers,

Tonya

---------------------------------------------------------
Dr Tonya Haff
Collection Manager
Australian National Wildlife Collection
National Research Collections Australia, CSIRO
Canberra, Australia
Phone: (+61) 02 6242 1566 (office)
(+61) 0419 569 109 (mobile)

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Subject: Digest Footer

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Nhcoll-l mailing list
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_______________________________________________
NHCOLL-L is brought to you by the Society for the Preservation of
Natural History Collections (SPNHC), an international society whose
mission is to improve the preservation, conservation and management of
natural history collections to ensure their continuing value to
society. See http://www.spnhc.org <http://www.spnhc.org%20> for membership information.
Advertising on NH-COLL-L is inappropriate.

------------------------------

End of Nhcoll-l Digest, Vol 108, Issue 4
****************************************
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