Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp
Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vi...
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| Veröffentlicht in: | Parasitology 2020-05, Vol.147 (6), p.659-666 |
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| creator | Scare, J A Dini, P Norris, J K Steuer, A E Scoggin, K Gravatte, H S Howe, D K Slusarewicz, P Nielsen, M K |
| description | Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp. |
| doi_str_mv | 10.1017/S0031182020000189 |
| format | Article |
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Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp.</description><identifier>ISSN: 0031-1820</identifier><identifier>EISSN: 1469-8161</identifier><identifier>DOI: 10.1017/S0031182020000189</identifier><identifier>PMID: 32046800</identifier><language>eng</language><publisher>England: Cambridge University Press</publisher><subject>Animals ; Anthelmintic agents ; Anthelmintics - pharmacology ; Antiparasitic agents ; Ascaridoidea - drug effects ; Ascaridoidea - genetics ; Detoxification ; Drug resistance ; Drug Resistance - genetics ; Exposure ; Gene Expression ; Gene Expression Profiling ; Genes, Helminth ; Genomes ; In vitro methods and tests ; In Vitro Techniques ; Ivermectin ; Metabolism ; Motility ; Parascaris ; Parasites ; Regulatory mechanisms (biology) ; Researchers ; Ribonucleic acid ; RNA ; RNA-Seq ; Transcriptome ; Transcriptomics ; Worms</subject><ispartof>Parasitology, 2020-05, Vol.147 (6), p.659-666</ispartof><rights>Copyright © Cambridge University Press 2020</rights><rights>Cambridge University Press 2020 2020 Cambridge University Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c428t-1f8963a01b87d8e0082f820bc80b6fed4f4cec54e9821a2bf8402dc406e1f4ee3</citedby><cites>FETCH-LOGICAL-c428t-1f8963a01b87d8e0082f820bc80b6fed4f4cec54e9821a2bf8402dc406e1f4ee3</cites><orcidid>0000-0001-9309-9879</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10317622/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10317622/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32046800$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Scare, J A</creatorcontrib><creatorcontrib>Dini, P</creatorcontrib><creatorcontrib>Norris, J K</creatorcontrib><creatorcontrib>Steuer, A E</creatorcontrib><creatorcontrib>Scoggin, K</creatorcontrib><creatorcontrib>Gravatte, H S</creatorcontrib><creatorcontrib>Howe, D K</creatorcontrib><creatorcontrib>Slusarewicz, P</creatorcontrib><creatorcontrib>Nielsen, M K</creatorcontrib><title>Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp</title><title>Parasitology</title><addtitle>Parasitology</addtitle><description>Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp.</description><subject>Animals</subject><subject>Anthelmintic agents</subject><subject>Anthelmintics - pharmacology</subject><subject>Antiparasitic agents</subject><subject>Ascaridoidea - drug effects</subject><subject>Ascaridoidea - genetics</subject><subject>Detoxification</subject><subject>Drug resistance</subject><subject>Drug Resistance - genetics</subject><subject>Exposure</subject><subject>Gene Expression</subject><subject>Gene Expression Profiling</subject><subject>Genes, Helminth</subject><subject>Genomes</subject><subject>In vitro methods and tests</subject><subject>In Vitro Techniques</subject><subject>Ivermectin</subject><subject>Metabolism</subject><subject>Motility</subject><subject>Parascaris</subject><subject>Parasites</subject><subject>Regulatory mechanisms (biology)</subject><subject>Researchers</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA-Seq</subject><subject>Transcriptome</subject><subject>Transcriptomics</subject><subject>Worms</subject><issn>0031-1820</issn><issn>1469-8161</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNplkVFvFCEQx4mxsWf1A_hiSHzpy9qBpRzni2kabU2a1ER9JiwMdzS7sMLupf1Ufgi_mJxXm6q8QOb_m_8wM4S8YvCWAVuefAFoGVMcONTD1OoJWTAhV41ikj0li53c7PRD8ryUm8rIVvJn5LDlIKQCWJDbs2JNDq5QvB1TQfeOGjrgtEmO-pRpiHQbppyoy_N6z8wZqYmOrjHiLpKxlJBijZn-roRCk6_vaYP9EOIULI3m548t0s8mm9_FCi3j-IIceNMXfHl_H5FvHz98Pb9srq4vPp2fXTVWcDU1zKuVbA2wTi2dQgDFfW2oswo66dEJLyzaU4ErxZnhnVcCuLMCJDIvENsj8n7vO87dgM5inLLp9ZjDYPKdTibov5UYNnqdtprV4S0l59Xh-N4hp-8zlkkPoVjsexMxzUXz9lQwyWEpK_rmH_QmzbnOpVICQCjgklWK7SmbUykZ_cNvGOjdYvV_i605rx-38ZDxZ5PtL_bkoIs</recordid><startdate>20200501</startdate><enddate>20200501</enddate><creator>Scare, J A</creator><creator>Dini, P</creator><creator>Norris, J K</creator><creator>Steuer, A E</creator><creator>Scoggin, K</creator><creator>Gravatte, H S</creator><creator>Howe, D K</creator><creator>Slusarewicz, P</creator><creator>Nielsen, M K</creator><general>Cambridge University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TM</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-9309-9879</orcidid></search><sort><creationdate>20200501</creationdate><title>Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp</title><author>Scare, J A ; Dini, P ; Norris, J K ; Steuer, A E ; Scoggin, K ; Gravatte, H S ; Howe, D K ; Slusarewicz, P ; Nielsen, M K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c428t-1f8963a01b87d8e0082f820bc80b6fed4f4cec54e9821a2bf8402dc406e1f4ee3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Anthelmintic agents</topic><topic>Anthelmintics - pharmacology</topic><topic>Antiparasitic agents</topic><topic>Ascaridoidea - drug effects</topic><topic>Ascaridoidea - genetics</topic><topic>Detoxification</topic><topic>Drug resistance</topic><topic>Drug Resistance - genetics</topic><topic>Exposure</topic><topic>Gene Expression</topic><topic>Gene Expression Profiling</topic><topic>Genes, Helminth</topic><topic>Genomes</topic><topic>In vitro methods and tests</topic><topic>In Vitro Techniques</topic><topic>Ivermectin</topic><topic>Metabolism</topic><topic>Motility</topic><topic>Parascaris</topic><topic>Parasites</topic><topic>Regulatory mechanisms (biology)</topic><topic>Researchers</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA-Seq</topic><topic>Transcriptome</topic><topic>Transcriptomics</topic><topic>Worms</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Scare, J A</creatorcontrib><creatorcontrib>Dini, P</creatorcontrib><creatorcontrib>Norris, J K</creatorcontrib><creatorcontrib>Steuer, A E</creatorcontrib><creatorcontrib>Scoggin, K</creatorcontrib><creatorcontrib>Gravatte, H S</creatorcontrib><creatorcontrib>Howe, D K</creatorcontrib><creatorcontrib>Slusarewicz, P</creatorcontrib><creatorcontrib>Nielsen, M K</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Scare, J A</au><au>Dini, P</au><au>Norris, J K</au><au>Steuer, A E</au><au>Scoggin, K</au><au>Gravatte, H S</au><au>Howe, D K</au><au>Slusarewicz, P</au><au>Nielsen, M K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp</atitle><jtitle>Parasitology</jtitle><addtitle>Parasitology</addtitle><date>2020-05-01</date><risdate>2020</risdate><volume>147</volume><issue>6</issue><spage>659</spage><epage>666</epage><pages>659-666</pages><issn>0031-1820</issn><eissn>1469-8161</eissn><abstract>Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp.</abstract><cop>England</cop><pub>Cambridge University Press</pub><pmid>32046800</pmid><doi>10.1017/S0031182020000189</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0001-9309-9879</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Parasitology, 2020-05, Vol.147 (6), p.659-666 |
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| subjects | Animals Anthelmintic agents Anthelmintics - pharmacology Antiparasitic agents Ascaridoidea - drug effects Ascaridoidea - genetics Detoxification Drug resistance Drug Resistance - genetics Exposure Gene Expression Gene Expression Profiling Genes, Helminth Genomes In vitro methods and tests In Vitro Techniques Ivermectin Metabolism Motility Parascaris Parasites Regulatory mechanisms (biology) Researchers Ribonucleic acid RNA RNA-Seq Transcriptome Transcriptomics Worms |
| title | Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp |
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