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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">I would agree completely with this summation after seeing Tom’s explanation. Ideally, tissue should be retaken from these specimens and stored according to best
practices in cold storage. There is anecdotal evidence that both room temperature and long term alcohol storage degrade DNA over time even though there is very little published work on this. You could then either tag each specimen with a unique tissue number
to tie vouchers to tissues or store them in separate vials. This way there would be no worries about future comtaimnation.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Andy<o:p></o:p></span></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-AU" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"> A : A : A :<br>
}<(((_°>.,.,.,.}<(((_°>.,.,.,.}<)))_°><br>
V V V<br>
Andy Bentley<br>
Ichthyology Collection Manager<br>
University of Kansas<br>
Biodiversity Institute<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-AU" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Dyche Hall<br>
<a href="x-apple-data-detectors://9"><span style="color:#0563C1">1345 Jayhawk Boulevard</span></a><br>
<a href="x-apple-data-detectors://9"><span style="color:#0563C1">Lawrence, KS, 66045-7561</span></a><br>
<a href="x-apple-data-detectors://9"><span style="color:#0563C1">USA</span></a><br>
<br>
Tel: <a href="tel:%28785%29%20864-3863" target="_blank"><span style="color:#0563C1">(785) 864-3863</span></a><br>
Fax: <a href="tel:%28785%29%20864-5335" target="_blank"><span style="color:#0563C1">(785) 864-5335</span></a> <br>
Email: <a href="mailto:abentley@ku.edu" target="_blank"><span style="color:#0563C1">abentley@ku.edu</span></a> </span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-AU" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><a href="http://ichthyology.biodiversity.ku.edu/" target="_blank"><span style="color:#0563C1">http://ichthyology.biodiversity.ku.edu</span></a></span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"> A : A : A :<br>
}<(((_°>.,.,.,.}<(((_°>.,.,.,.}<)))_°><br>
V V V<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> Nhcoll-l <nhcoll-l-bounces@mailman.yale.edu>
<b>On Behalf Of </b>Rob Robins<br>
<b>Sent:</b> Friday, February 22, 2019 9:50 AM<br>
<b>To:</b> Thomas Labedz <tlabedz1@unl.edu>; John E Simmons <simmons.johne@gmail.com><br>
<b>Cc:</b> NHCOLL-L@mailman.yale.edu<br>
<b>Subject:</b> Re: [Nhcoll-l] eDNA voucher question<o:p></o:p></span></p>
</div>
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<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Tom,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Yes, this is the classic collection manager’s conundrum of: expend labor now to save space or conserve labor now at the expense of space.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">It looks to me like you can satisfy both your concern over wasted space and the researcher’s concern regarding individual separation of the specimens by:<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoListParagraph" style="text-indent:-.25in;mso-list:l0 level1 lfo2"><![if !supportLists]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><span style="mso-list:Ignore">1.<span style="font:7.0pt "Times New Roman"">
</span></span></span><![endif]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Resampling each fish for dna.
<o:p></o:p></span></p>
<p class="MsoListParagraph" style="text-indent:-.25in;mso-list:l0 level1 lfo2"><![if !supportLists]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><span style="mso-list:Ignore">2.<span style="font:7.0pt "Times New Roman"">
</span></span></span><![endif]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Linking those tissue samples to the individual specimens through the use of unique sampling locations on the body or small tags.<o:p></o:p></span></p>
<p class="MsoListParagraph" style="text-indent:-.25in;mso-list:l0 level1 lfo2"><![if !supportLists]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><span style="mso-list:Ignore">3.<span style="font:7.0pt "Times New Roman"">
</span></span></span><![endif]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Archiving the tiny, appropriately labeled tissue vials in an ultracold.<o:p></o:p></span></p>
<p class="MsoListParagraph" style="text-indent:-.25in;mso-list:l0 level1 lfo2"><![if !supportLists]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><span style="mso-list:Ignore">4.<span style="font:7.0pt "Times New Roman"">
</span></span></span><![endif]><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Commingling your whole fish voucher specimens in locality/collecting event specific specimen lots (4 containers and not 35).<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Or in other words, just reproduce the usual protocol for fishes coming from the field to the collection. I’m no DNA specialist, but I would think sampling the tissues now and
storing them in the ultracold would also guard against room-temperature degradation of dna (I’m told this is a thing) in the specimens themselves and of course, give you that access to the whole specimens you desire.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Rob<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<div>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Robert H. Robins<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Collection Manager<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Division of Ichthyology<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><img border="0" width="155" height="56" style="width:1.6145in;height:.5833in" id="Picture_x0020_1" src="cid:image001.jpg@01D4CA9A.E138D700" alt="FLMNH Fishes logo email small"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Florida Museum of Natural History<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">1659 Museum Road<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Gainesville, FL 32611-7800<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Office: (352) 273-1957<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Fax: (352) 846-0287<o:p></o:p></span></p>
<p class="MsoNormal"><a href="mailto:rhrobins@flmnh.ufl.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#0563C1">rhrobins@flmnh.ufl.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p></o:p></span></p>
<p class="MsoNormal"><a href="https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.flmnh.ufl.edu%2F&data=02%7C01%7Cabentley%40ku.edu%7C5ebfc241f0db4bbb5fe908d698dd6f73%7C3c176536afe643f5b96636feabbe3c1a%7C0%7C0%7C636864474162815698&sdata=7sz1XnnAqrtru2bchDPaYP%2FiO3Ls7D23E9FUETMLWi4%3D&reserved=0"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#0563C1">www.flmnh.ufl.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><img border="0" width="227" height="324" style="width:2.3645in;height:3.375in" id="Picture_x0020_2" src="cid:image002.jpg@01D4CA9A.E138D700" alt="Fishes in the FWofF_cover_front resized 33"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">Search the Collection:
<o:p></o:p></span></p>
<p class="MsoNormal"><a href="https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fspecifyportal.flmnh.ufl.edu%2Ffishes%2F&data=02%7C01%7Cabentley%40ku.edu%7C5ebfc241f0db4bbb5fe908d698dd6f73%7C3c176536afe643f5b96636feabbe3c1a%7C0%7C0%7C636864474162825707&sdata=jDspV5MnOShIMhDYlL8F01ccvtTjeT9QnVJNM96fpbY%3D&reserved=0"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#0563C1">http://specifyportal.flmnh.ufl.edu/fishes/</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> Nhcoll-l <<a href="mailto:nhcoll-l-bounces@mailman.yale.edu">nhcoll-l-bounces@mailman.yale.edu</a>>
<b>On Behalf Of </b>Thomas Labedz<br>
<b>Sent:</b> Friday, February 22, 2019 10:31 AM<br>
<b>To:</b> John E Simmons <<a href="mailto:simmons.johne@gmail.com">simmons.johne@gmail.com</a>><br>
<b>Cc:</b> <a href="mailto:NHCOLL-L@mailman.yale.edu">NHCOLL-L@mailman.yale.edu</a><br>
<b>Subject:</b> Re: [Nhcoll-l] eDNA voucher question<o:p></o:p></span></p>
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</div>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">NHCOLL readers<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">In an attempt to clarify my question to another responder I’ve copied below a more thorough explanation. Thanks to you all for your time and knowledge.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Thomas Labedz<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">University of Nebraska State Museum<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">The DNA work was done at another university, one that does not have collections to maintain vouchers. The DNA was collected and analyzed from these vouchers, the paper written and submitted for publication, and
the author/researcher wants to deposit the individual fish as vouchers. Shoal Chub (<i>Macrhybopsis hyostoma</i>) are small, the largest maybe 5 cm long. My institution does not have any of the removed tissues that were analyzed for DNA, nor any products from
that DNA, only the fish themselves. The researcher’s preference is for individual containers for each specimen should someone want to extract new tissue for DNA. Doing so would mean an approximately 10X increase in shelf space and material necessary to maintain
these vouchers. Since we do not have the DNA samples or products, and are holding only the fish themselves, I submit that we should individually package each fish with its information but maintain all the samples from the same collecting locality in joint
jars (4 jars instead of 35). Yes, cross-contamination would potentially occur between those jointly held samples, but since they are from the same collecting site cross-contamination would also be true in the natural environment where they were collected.
Being small one option is to stuff them into individual tubes and put them in the freezers with tissues from other specimens where the vouchers are at this institution. That might work in this case but doesn’t lend itself to ready access for species verification,
nor does it function well as policy or protocol should another project produce larger numbers of samples from larger species.
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="color:#1F497D">Guess I’m addressing the balance between scientific need and budgetary/space constraints.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"> John E Simmons <</span><a href="mailto:simmons.johne@gmail.com"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">simmons.johne@gmail.com</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">>
<br>
<b>Sent:</b> Friday, February 22, 2019 9:26 AM<br>
<b>To:</b> Thomas Labedz <</span><a href="mailto:tlabedz1@unl.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">tlabedz1@unl.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">><br>
<b>Cc:</b> </span><a href="mailto:NHCOLL-L@mailman.yale.edu"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif">NHCOLL-L@mailman.yale.edu</span></a><span style="font-size:11.0pt;font-family:"Calibri",sans-serif"><br>
<b>Subject:</b> Re: [Nhcoll-l] eDNA voucher question<o:p></o:p></span></p>
<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">Tom,<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">I am not sure I understand the question. Do you mean the fish are vouchers from which DNA has been extracted and you want to keep the specimen separate to link it to the DNA sample,
or do you mean fish from which DNA will be extracted in the future? <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">You will get contamination of the alcohol around the specimen from DNA from other specimens if you house them all together because protein is extracted over time by the alcohol (for
example, see von Endt 1994, "Spirit collections: a preliminary analysis of some organic materials found in the storage fluid of mammals," Collection Forum 10(1):10-19). Whether that contamination would affect the DNA in the fish specimen itself has not been
looked at, to my knowledge, but is an interesting question.<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">If you are storing these as whole-specimen vouchers from which DNA has already been sampled, I would go with Andy's suggestion of vials with polyester fiber plugs. If you are storing
these for future DNA extraction, then they should be in 95.6% ETOH if they must be kept in alcohol.<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">Please let me know if I have misunderstood your question.<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-family:"Tahoma",sans-serif;color:black">--John<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif">John E. Simmons<br>
Writer and Museum Consultant</span><o:p></o:p></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif">Museologica<br>
<i>and</i><br>
Associate Curator of Collections<br>
Earth and Mineral Science Museum & Art Gallery<br>
Penn State University<br>
<i>and</i><br>
Investigador Asociado, Departamento de Ornitologia<br>
Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima</span><o:p></o:p></p>
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<p class="MsoNormal">On Thu, Feb 21, 2019 at 4:41 PM Thomas Labedz <<a href="mailto:tlabedz1@unl.edu">tlabedz1@unl.edu</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Greetings from snowy central North America!<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Has anyone developed policy or protocol for the receipt, treatment, and maintenance of eDNA fish voucher specimens? Would you share? The individual fish are small and the researcher
recommends keeping each individual in a separate container (how they were delivered) to prohibit cross contamination of DNA via the ethanol solution. That is understandable but will utilize lots of space and containers.<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Thomas<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">Thomas E. Labedz, Collections Manager</span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">Division of Zoology and Division of Botany</span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">University of Nebraska State Museum</span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">Lincoln, Nebraska, U.S.A.</span><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><a href="mailto:tlabedz1@unl.edu" target="_blank"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">tlabedz1@unl.edu</span></a><span style="font-size:10.0pt;font-family:"Arial",sans-serif">
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"> <o:p></o:p></p>
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