Loading…

Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling

Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were to describe an inexpensive and effective two-step euthanasia m...

Full description

Saved in:
Bibliographic Details
Published in:Animals (Basel) 2021-06, Vol.11 (7), p.1847
Main Authors: Wahltinez, Sarah J., Kroll, Kevin J., Nunamaker, Elizabeth A., Denslow, Nancy D., Stacy, Nicole I.
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3
cites cdi_FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3
container_end_page
container_issue 7
container_start_page 1847
container_title Animals (Basel)
container_volume 11
creator Wahltinez, Sarah J.
Kroll, Kevin J.
Nunamaker, Elizabeth A.
Denslow, Nancy D.
Stacy, Nicole I.
description Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were to describe an inexpensive and effective two-step euthanasia method for adult common sea stars (Asterias rubens) and to demonstrate that high-quality RNA samples for further use in downstream molecular analyses can be obtained from pyloric ceca of MgCl2-immersed sea stars. Adult common sea stars (n = 15) were immersed in a 75 g/L magnesium chloride solution until they were no longer reactive to having their oral surface tapped with forceps (mean: 4 min, range 2–7 min), left immersed for an additional minute, and then sampled with sharp scissors. RNA from pyloric ceca (n = 10) was isolated using a liquid–liquid method, then samples were treated with DNase and analyzed for evaluation of RNA integrity number (RIN) for assessment of the quantity and purity of intact RNA. Aversive reactions to magnesium chloride solution were not observed and no sea stars regained spontaneous movement or reacted to sampling. The calculated RIN ranged from 7.3–9.8, demonstrating that the combination of animal welfare via the use of anesthesia and sampling for advanced molecular techniques is possible using this low-cost technique.
doi_str_mv 10.3390/ani11071847
format article
fullrecord <record><control><sourceid>proquest_doaj_</sourceid><recordid>TN_cdi_doaj_primary_oai_doaj_org_article_7e8eaf10e00841d6a0861d73bd1f8de8</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><doaj_id>oai_doaj_org_article_7e8eaf10e00841d6a0861d73bd1f8de8</doaj_id><sourcerecordid>2554361235</sourcerecordid><originalsourceid>FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3</originalsourceid><addsrcrecordid>eNpdkktr3DAQgE1paUKaU_-AoJeU4lYP6-FLYVnSJpA-Nz2LsSSvtcjWRpJb8u_rZENJqsuImU8fM2Kq6jXB7xlr8QeYPCFYEtXIZ9UxxVLUVBD-_NH9qDrNeYeXIzkjnLysjlhDsaBNe1yF7wlM8QYCOp_LABNkD-iLK0O0qI8JreM4xgltHKBNgZTR2SoXlzxklObOTfktuh6goFUI8U--f3Lht0P9Y4bgyy36-XWFNjDug5-2r6oXPYTsTh_iSfXr0_n1-qK--vb5cr26qk3Daak7Sh2jUrW8aRTjlhgraMe4WHK96ClRbcNs1zln2pa02Bojcd8JyxV3wICdVJcHr42w0_vkR0i3OoLX94mYthrSMnRwWjrloCfYYawaYgVgJYiVrLOkV9apxfXx4NrP3eiscVNJEJ5In1YmP-ht_K0Vw5i1chGcPQhSvJldLnr02bgQYHJxzpryZchWSHaHvvkP3cU5TctXLRRvmCCU8YV6d6BMijkn1_9rhmB9txT60VKwv4ztp3E</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2554361235</pqid></control><display><type>article</type><title>Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling</title><source>Publicly Available Content Database (Proquest) (PQ_SDU_P3)</source><source>PubMed Central Free</source><creator>Wahltinez, Sarah J. ; Kroll, Kevin J. ; Nunamaker, Elizabeth A. ; Denslow, Nancy D. ; Stacy, Nicole I.</creator><creatorcontrib>Wahltinez, Sarah J. ; Kroll, Kevin J. ; Nunamaker, Elizabeth A. ; Denslow, Nancy D. ; Stacy, Nicole I.</creatorcontrib><description>Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were to describe an inexpensive and effective two-step euthanasia method for adult common sea stars (Asterias rubens) and to demonstrate that high-quality RNA samples for further use in downstream molecular analyses can be obtained from pyloric ceca of MgCl2-immersed sea stars. Adult common sea stars (n = 15) were immersed in a 75 g/L magnesium chloride solution until they were no longer reactive to having their oral surface tapped with forceps (mean: 4 min, range 2–7 min), left immersed for an additional minute, and then sampled with sharp scissors. RNA from pyloric ceca (n = 10) was isolated using a liquid–liquid method, then samples were treated with DNase and analyzed for evaluation of RNA integrity number (RIN) for assessment of the quantity and purity of intact RNA. Aversive reactions to magnesium chloride solution were not observed and no sea stars regained spontaneous movement or reacted to sampling. The calculated RIN ranged from 7.3–9.8, demonstrating that the combination of animal welfare via the use of anesthesia and sampling for advanced molecular techniques is possible using this low-cost technique.</description><identifier>ISSN: 2076-2615</identifier><identifier>EISSN: 2076-2615</identifier><identifier>DOI: 10.3390/ani11071847</identifier><identifier>PMID: 34206249</identifier><language>eng</language><publisher>Basel: MDPI AG</publisher><subject>Alkalinity ; Ammonia ; Anesthesia ; Animal behavior ; Animal tissues ; Animal welfare ; Aquariums ; Asterias rubens ; Biodiversity ; Cutting tools ; Deoxyribonuclease ; Digestive glands ; Euthanasia ; invertebrate ; Invertebrates ; killing ; Laboratory animals ; Magnesium ; Magnesium chloride ; Marine ecosystems ; Marine invertebrates ; Medical instruments ; Methods ; Salinity ; Sampling ; sea star ; starfish ; Water quality</subject><ispartof>Animals (Basel), 2021-06, Vol.11 (7), p.1847</ispartof><rights>2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 by the authors. 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3</citedby><cites>FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3</cites><orcidid>0000-0003-4258-4414 ; 0000-0002-4232-4420 ; 0000-0001-7639-4454</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2554361235/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2554361235?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids></links><search><creatorcontrib>Wahltinez, Sarah J.</creatorcontrib><creatorcontrib>Kroll, Kevin J.</creatorcontrib><creatorcontrib>Nunamaker, Elizabeth A.</creatorcontrib><creatorcontrib>Denslow, Nancy D.</creatorcontrib><creatorcontrib>Stacy, Nicole I.</creatorcontrib><title>Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling</title><title>Animals (Basel)</title><description>Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were to describe an inexpensive and effective two-step euthanasia method for adult common sea stars (Asterias rubens) and to demonstrate that high-quality RNA samples for further use in downstream molecular analyses can be obtained from pyloric ceca of MgCl2-immersed sea stars. Adult common sea stars (n = 15) were immersed in a 75 g/L magnesium chloride solution until they were no longer reactive to having their oral surface tapped with forceps (mean: 4 min, range 2–7 min), left immersed for an additional minute, and then sampled with sharp scissors. RNA from pyloric ceca (n = 10) was isolated using a liquid–liquid method, then samples were treated with DNase and analyzed for evaluation of RNA integrity number (RIN) for assessment of the quantity and purity of intact RNA. Aversive reactions to magnesium chloride solution were not observed and no sea stars regained spontaneous movement or reacted to sampling. The calculated RIN ranged from 7.3–9.8, demonstrating that the combination of animal welfare via the use of anesthesia and sampling for advanced molecular techniques is possible using this low-cost technique.</description><subject>Alkalinity</subject><subject>Ammonia</subject><subject>Anesthesia</subject><subject>Animal behavior</subject><subject>Animal tissues</subject><subject>Animal welfare</subject><subject>Aquariums</subject><subject>Asterias rubens</subject><subject>Biodiversity</subject><subject>Cutting tools</subject><subject>Deoxyribonuclease</subject><subject>Digestive glands</subject><subject>Euthanasia</subject><subject>invertebrate</subject><subject>Invertebrates</subject><subject>killing</subject><subject>Laboratory animals</subject><subject>Magnesium</subject><subject>Magnesium chloride</subject><subject>Marine ecosystems</subject><subject>Marine invertebrates</subject><subject>Medical instruments</subject><subject>Methods</subject><subject>Salinity</subject><subject>Sampling</subject><subject>sea star</subject><subject>starfish</subject><subject>Water quality</subject><issn>2076-2615</issn><issn>2076-2615</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNpdkktr3DAQgE1paUKaU_-AoJeU4lYP6-FLYVnSJpA-Nz2LsSSvtcjWRpJb8u_rZENJqsuImU8fM2Kq6jXB7xlr8QeYPCFYEtXIZ9UxxVLUVBD-_NH9qDrNeYeXIzkjnLysjlhDsaBNe1yF7wlM8QYCOp_LABNkD-iLK0O0qI8JreM4xgltHKBNgZTR2SoXlzxklObOTfktuh6goFUI8U--f3Lht0P9Y4bgyy36-XWFNjDug5-2r6oXPYTsTh_iSfXr0_n1-qK--vb5cr26qk3Daak7Sh2jUrW8aRTjlhgraMe4WHK96ClRbcNs1zln2pa02Bojcd8JyxV3wICdVJcHr42w0_vkR0i3OoLX94mYthrSMnRwWjrloCfYYawaYgVgJYiVrLOkV9apxfXx4NrP3eiscVNJEJ5In1YmP-ht_K0Vw5i1chGcPQhSvJldLnr02bgQYHJxzpryZchWSHaHvvkP3cU5TctXLRRvmCCU8YV6d6BMijkn1_9rhmB9txT60VKwv4ztp3E</recordid><startdate>20210622</startdate><enddate>20210622</enddate><creator>Wahltinez, Sarah J.</creator><creator>Kroll, Kevin J.</creator><creator>Nunamaker, Elizabeth A.</creator><creator>Denslow, Nancy D.</creator><creator>Stacy, Nicole I.</creator><general>MDPI AG</general><general>MDPI</general><scope>AAYXX</scope><scope>CITATION</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-4258-4414</orcidid><orcidid>https://orcid.org/0000-0002-4232-4420</orcidid><orcidid>https://orcid.org/0000-0001-7639-4454</orcidid></search><sort><creationdate>20210622</creationdate><title>Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling</title><author>Wahltinez, Sarah J. ; Kroll, Kevin J. ; Nunamaker, Elizabeth A. ; Denslow, Nancy D. ; Stacy, Nicole I.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Alkalinity</topic><topic>Ammonia</topic><topic>Anesthesia</topic><topic>Animal behavior</topic><topic>Animal tissues</topic><topic>Animal welfare</topic><topic>Aquariums</topic><topic>Asterias rubens</topic><topic>Biodiversity</topic><topic>Cutting tools</topic><topic>Deoxyribonuclease</topic><topic>Digestive glands</topic><topic>Euthanasia</topic><topic>invertebrate</topic><topic>Invertebrates</topic><topic>killing</topic><topic>Laboratory animals</topic><topic>Magnesium</topic><topic>Magnesium chloride</topic><topic>Marine ecosystems</topic><topic>Marine invertebrates</topic><topic>Medical instruments</topic><topic>Methods</topic><topic>Salinity</topic><topic>Sampling</topic><topic>sea star</topic><topic>starfish</topic><topic>Water quality</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wahltinez, Sarah J.</creatorcontrib><creatorcontrib>Kroll, Kevin J.</creatorcontrib><creatorcontrib>Nunamaker, Elizabeth A.</creatorcontrib><creatorcontrib>Denslow, Nancy D.</creatorcontrib><creatorcontrib>Stacy, Nicole I.</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Publicly Available Content Database (Proquest) (PQ_SDU_P3)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>Directory of Open Access Journals</collection><jtitle>Animals (Basel)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wahltinez, Sarah J.</au><au>Kroll, Kevin J.</au><au>Nunamaker, Elizabeth A.</au><au>Denslow, Nancy D.</au><au>Stacy, Nicole I.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling</atitle><jtitle>Animals (Basel)</jtitle><date>2021-06-22</date><risdate>2021</risdate><volume>11</volume><issue>7</issue><spage>1847</spage><pages>1847-</pages><issn>2076-2615</issn><eissn>2076-2615</eissn><abstract>Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were to describe an inexpensive and effective two-step euthanasia method for adult common sea stars (Asterias rubens) and to demonstrate that high-quality RNA samples for further use in downstream molecular analyses can be obtained from pyloric ceca of MgCl2-immersed sea stars. Adult common sea stars (n = 15) were immersed in a 75 g/L magnesium chloride solution until they were no longer reactive to having their oral surface tapped with forceps (mean: 4 min, range 2–7 min), left immersed for an additional minute, and then sampled with sharp scissors. RNA from pyloric ceca (n = 10) was isolated using a liquid–liquid method, then samples were treated with DNase and analyzed for evaluation of RNA integrity number (RIN) for assessment of the quantity and purity of intact RNA. Aversive reactions to magnesium chloride solution were not observed and no sea stars regained spontaneous movement or reacted to sampling. The calculated RIN ranged from 7.3–9.8, demonstrating that the combination of animal welfare via the use of anesthesia and sampling for advanced molecular techniques is possible using this low-cost technique.</abstract><cop>Basel</cop><pub>MDPI AG</pub><pmid>34206249</pmid><doi>10.3390/ani11071847</doi><orcidid>https://orcid.org/0000-0003-4258-4414</orcidid><orcidid>https://orcid.org/0000-0002-4232-4420</orcidid><orcidid>https://orcid.org/0000-0001-7639-4454</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 2076-2615
ispartof Animals (Basel), 2021-06, Vol.11 (7), p.1847
issn 2076-2615
2076-2615
language eng
recordid cdi_doaj_primary_oai_doaj_org_article_7e8eaf10e00841d6a0861d73bd1f8de8
source Publicly Available Content Database (Proquest) (PQ_SDU_P3); PubMed Central Free
subjects Alkalinity
Ammonia
Anesthesia
Animal behavior
Animal tissues
Animal welfare
Aquariums
Asterias rubens
Biodiversity
Cutting tools
Deoxyribonuclease
Digestive glands
Euthanasia
invertebrate
Invertebrates
killing
Laboratory animals
Magnesium
Magnesium chloride
Marine ecosystems
Marine invertebrates
Medical instruments
Methods
Salinity
Sampling
sea star
starfish
Water quality
title Practical Euthanasia Method for Common Sea Stars (Asterias rubens) That Allows for High-Quality RNA Sampling
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-26T01%3A02%3A20IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_doaj_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Practical%20Euthanasia%20Method%20for%20Common%20Sea%20Stars%20(Asterias%20rubens)%20That%20Allows%20for%20High-Quality%20RNA%20Sampling&rft.jtitle=Animals%20(Basel)&rft.au=Wahltinez,%20Sarah%20J.&rft.date=2021-06-22&rft.volume=11&rft.issue=7&rft.spage=1847&rft.pages=1847-&rft.issn=2076-2615&rft.eissn=2076-2615&rft_id=info:doi/10.3390/ani11071847&rft_dat=%3Cproquest_doaj_%3E2554361235%3C/proquest_doaj_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c452t-b22e32789544835d1cd62b356278f6f218943dbbeec99190dcc70fb6d585ea3a3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2554361235&rft_id=info:pmid/34206249&rfr_iscdi=true