{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T18:05:54Z","timestamp":1787162754233,"version":"build-2736575974"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2020,8,6]],"date-time":"2020-08-06T00:00:00Z","timestamp":1596672000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100001458","name":"American Kennel Club Canine Health Foundation","doi-asserted-by":"publisher","award":["02458-A"],"award-info":[{"award-number":["02458-A"]}],"id":[{"id":"10.13039\/100001458","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R21AI144493-02"],"award-info":[{"award-number":["5R21AI144493-02"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["2017-04619"],"award-info":[{"award-number":["2017-04619"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"DOI":"10.1371\/journal.pone.0237150","type":"journal-article","created":{"date-parts":[[2020,8,6]],"date-time":"2020-08-06T15:02:35Z","timestamp":1596726155000},"page":"e0237150","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["Laboratory assays reveal diverse phenotypes among microfilariae of Dirofilaria immitis isolates with known macrocyclic lactone susceptibility status"],"prefix":"10.1371","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3077-0201","authenticated-orcid":true,"given":"Jeba R. J.","family":"Jesudoss Chelladurai","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katy A.","family":"Martin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0679-3542","authenticated-orcid":true,"given":"Krystal","family":"Chinchilla-Vargas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5596-7380","authenticated-orcid":true,"given":"Pablo D.","family":"Jimenez Castro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ray M.","family":"Kaplan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7538-1934","authenticated-orcid":true,"given":"Matthew T.","family":"Brewer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2020,8,6]]},"reference":[{"key":"pone.0237150.ref001","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1017\/S003118201500061X","article-title":"The emergence of macrocyclic lactone resistance in the canine heartworm, Dirofilaria immitis","volume":"142","author":"AJ Wolstenholme","year":"2015","journal-title":"Parasitology"},{"key":"pone.0237150.ref002","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.vetpar.2005.04.004","article-title":"Evaluation of efficacy of heartworm preventive products at the FDA","volume":"133","author":"VA Hampshire","year":"2005","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref003","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1186\/s13071-014-0494-6","article-title":"Establishment of macrocyclic lactone resistant Dirofilaria immitis isolates in experimentally infected laboratory dogs","volume":"7","author":"CN Pulaski","year":"2014","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref004","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1186\/s13071-017-2441-9","article-title":"A diagnostic algorithm for evaluating cases of potential macrocyclic lactone-resistant heartworm","volume":"10","author":"AR Moorhead","year":"2017","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref005","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1053\/j.tcam.2011.09.004","article-title":"Evidence for macrocyclic lactone anthelmintic resistance in Dirofilaria immitis","volume":"26","author":"TG Geary","year":"2011","journal-title":"Top Companion Anim Med"},{"key":"pone.0237150.ref006","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.ijpddr.2013.05.001","article-title":"Development of an in vitro bioassay for measuring susceptibility to macrocyclic lactone anthelmintics in Dirofilaria immitis","volume":"3","author":"CC Evans","year":"2013","journal-title":"Int J Parasitol Drugs Drug Resist"},{"key":"pone.0237150.ref007","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.vetpar.2017.09.003","article-title":"Evaluation of the larval migration inhibition assay for detecting macrocyclic lactone resistance in Dirofilaria immitis","volume":"246","author":"CC Evans","year":"2017","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref008","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1186\/s13071-017-2436-6","article-title":"Does evaluation of in vitro microfilarial motility reflect the resistance status of Dirofilaria immitis isolates to macrocyclic lactones?","volume":"10","author":"MJ Maclean","year":"2017","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref009","doi-asserted-by":"crossref","first-page":"596","DOI":"10.1016\/j.ijpddr.2018.06.006","article-title":"Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis","volume":"8","author":"C Ballesteros","year":"2018","journal-title":"Int J Parasitol Drugs Drug Resist"},{"key":"pone.0237150.ref010","doi-asserted-by":"crossref","first-page":"e70702","DOI":"10.1371\/journal.pone.0070702","article-title":"An extensive comparison of the effect of anthelmintic classes on diverse nematodes","volume":"8","author":"Y Hu","year":"2013","journal-title":"PLoS One"},{"key":"pone.0237150.ref011","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.vetpar.2014.02.004","article-title":"Ivermectin-dependent attachment of neutrophils and peripheral blood mononuclear cells to Dirofilaria immitis microfilariae in vitro","volume":"206","author":"AF Vatta","year":"2014","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref012","doi-asserted-by":"crossref","first-page":"4809","DOI":"10.1128\/AAC.00713-16","article-title":"Acquired Tolerance to Ivermectin and Moxidectin after Drug Selection Pressure in the Nematode Caenorhabditis elegans","volume":"60","author":"C M\u00e9nez","year":"2016","journal-title":"Antimicrob Agents Chemother"},{"key":"pone.0237150.ref013","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1186\/s13071-016-1806-9","article-title":"Increased expression of ATP binding cassette transporter genes following exposure of Haemonchus contortus larvae to a high concentration of monepantel in vitro","volume":"9","author":"A Raza","year":"2016","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref014","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.vetpar.2003.09.016","article-title":"Evaluation of a larval development assay (DrenchRite) for the detection of anthelmintic resistance in cyathostomin nematodes of horses","volume":"121","author":"R Tandon","year":"2004","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref015","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1111\/pim.12037","article-title":"Automated imaging and other developments in whole-organism anthelmintic screening","volume":"35","author":"RA Paveley","year":"2013","journal-title":"Parasite Immunol"},{"key":"pone.0237150.ref016","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1590\/S0001-37652011000200024","article-title":"Cross-disciplinary approaches for measuring parasitic helminth viability and phenotype","volume":"83","author":"E Peak","year":"2011","journal-title":"An Acad Bras Cienc"},{"key":"pone.0237150.ref017","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1128\/AAC.1.6.451","article-title":"Standardized single-disc method for antibiotic susceptibility testing of anaerobic bacteria","volume":"1","author":"TD Wikins","year":"1972","journal-title":"Antimicrob Agents Chemother"},{"key":"pone.0237150.ref018","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.vetpar.2011.01.024","article-title":"Correlation between loss of efficacy of macrocyclic lactone heartworm anthelmintics and P-glycoprotein genotype","volume":"176","author":"C Bourguinat","year":"2011","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref019","first-page":"70","article-title":"A method for staining nematode secretions and structures","volume":"20","author":"D Premachandran","year":"1988","journal-title":"J Nematol"},{"key":"pone.0237150.ref020","doi-asserted-by":"crossref","first-page":"879263","DOI":"10.1155\/2015\/879263","article-title":"A new methodology for evaluation of nematode viability","volume":"2015","author":"SR Ferreira","year":"2015","journal-title":"Biomed Res Int"},{"key":"pone.0237150.ref021","doi-asserted-by":"crossref","first-page":"3280","DOI":"10.1016\/j.fct.2012.06.051","article-title":"Toxicity ranking of heavy metals with screening method using adult Caenorhabditis elegans and propidium iodide replicates toxicity ranking in rat","volume":"50","author":"PR Hunt","year":"2012","journal-title":"Food Chem Toxicol"},{"key":"pone.0237150.ref022","doi-asserted-by":"crossref","first-page":"e795","DOI":"10.1371\/journal.pntd.0000795","article-title":"Comparison of microscopy and Alamar blue reduction in a larval based assay for schistosome drug screening","volume":"4","author":"NR Mansour","year":"2010","journal-title":"PLoS Negl Trop Dis"},{"key":"pone.0237150.ref023","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.mimet.2011.08.009","article-title":"Exploration of novel in vitro assays to study drugs against Trichuris spp","volume":"87","author":"A Silbereisen","year":"2011","journal-title":"J Microbiol Methods"},{"key":"pone.0237150.ref024","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1007\/s00436-007-0572-1","article-title":"A rapid colorimetric assay for the quantitation of the viability of free-living larvae of nematodes in vitro","volume":"101","author":"CE James","year":"2007","journal-title":"Parasitol Res"},{"key":"pone.0237150.ref025","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/0020-7519(89)90024-6","article-title":"Colorimetric quantitation of filarial viability","volume":"19","author":"JC Comley","year":"1989","journal-title":"Int J Parasitol"},{"key":"pone.0237150.ref026","doi-asserted-by":"crossref","DOI":"10.21769\/BioProtoc.2048","article-title":"Intracellular Assessment of ATP Levels in","volume":"6","author":"K Palikaras","year":"2016","journal-title":"Bio Protoc"},{"key":"pone.0237150.ref027","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.exppara.2017.07.008","article-title":"Assessment of in vitro killing assays for detecting praziquantel-induced death in Posthodiplostomum minimum metacercariae","volume":"181","author":"C Bader","year":"2017","journal-title":"Experimental Parasitology"},{"key":"pone.0237150.ref028","doi-asserted-by":"crossref","first-page":"2224","DOI":"10.1128\/AAC.01477-10","article-title":"Anthelmintics are substrates and activators of nematode P glycoprotein","volume":"55","author":"D Kerboeuf","year":"2011","journal-title":"Antimicrob Agents Chemother"},{"key":"pone.0237150.ref029","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/s13071-018-2631-0","article-title":"Increasing incidence of Dirofilaria immitis in dogs in USA with focus on the southeast region 2013\u20132016","volume":"11","author":"J Drake","year":"2018","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref030","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1186\/s13071-019-3633-2","article-title":"Regional and local temporal trends in the prevalence of canine heartworm infection in the contiguous United States: 2012\u20132018","volume":"12","author":"SW Self","year":"2019","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref031","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/j.ijpara.2016.04.004","article-title":"Interaction of macrocyclic lactones with a Dirofilaria immitis P-glycoprotein","volume":"46","author":"T Mani","year":"2016","journal-title":"Int J Parasitol"},{"key":"pone.0237150.ref032","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1186\/s13071-019-3645-y","article-title":"Differential ABC transporter gene expression in adult Dirofilaria immitis males and females following in vitro treatment with ivermectin, doxycycline or a combination of both","volume":"12","author":"C Lucchetti","year":"2019","journal-title":"Parasit Vectors"},{"key":"pone.0237150.ref033","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.vetpar.2019.02.002","article-title":"Detection and quantification of Parascaris P-glycoprotein drug transporter expression with a novel mRNA hybridization technique","volume":"267","author":"J Jesudoss Chelladurai","year":"2019","journal-title":"Vet Parasitol"},{"key":"pone.0237150.ref034","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1645\/0022-3395(2002)088[0612:LOPGMI]2.0.CO;2","article-title":"Localization of p-glycoprotein mRNA in the tissues of Haemonchus contortus adult worms and its relative abundance in drug-selected and susceptible strains","volume":"88","author":"JM Smith","year":"2002","journal-title":"J Parasitol"},{"key":"pone.0237150.ref035","doi-asserted-by":"crossref","first-page":"e22429","DOI":"10.1371\/journal.pone.0022429","article-title":"Sensitive and specific fluorescent probes for functional analysis of the three major types of mammalian ABC transporters","volume":"6","author":"IV Lebedeva","year":"2011","journal-title":"PLoS One"},{"key":"pone.0237150.ref036","doi-asserted-by":"crossref","first-page":"1747","DOI":"10.1021\/ci400195v","article-title":"Molecular docking characterizes substrate-binding sites and efflux modulation mechanisms within P-glycoprotein","volume":"53","author":"RJ Ferreira","year":"2013","journal-title":"J Chem Inf Model"},{"key":"pone.0237150.ref037","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.ijpddr.2016.04.001","article-title":"ABC-B transporter genes in Dirofilaria immitis","volume":"6","author":"C Bourguinat","year":"2016","journal-title":"Int J Parasitol Drugs Drug Resist"},{"key":"pone.0237150.ref038","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1002\/jps.20046","article-title":"Bidirectional transport of rhodamine 123 and Hoechst 33342, fluorescence probes of the binding sites on P-glycoprotein, across MDCK-MDR1 cell monolayers","volume":"93","author":"F Tang","year":"2004","journal-title":"J Pharm Sci"},{"key":"pone.0237150.ref039","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1111\/jvp.12330","article-title":"Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance","volume":"40","author":"JH Whittaker","year":"2017","journal-title":"J Vet Pharmacol Ther"},{"key":"pone.0237150.ref040","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.molbiopara.2011.09.003","article-title":"Candidate anthelmintic resistance-associated gene expression and sequence polymorphisms in a triple-resistant field isolate of Haemonchus contortus","volume":"180","author":"SM Williamson","year":"2011","journal-title":"Mol Biochem Parasitol"},{"key":"pone.0237150.ref041","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1017\/S0022149X11000216","article-title":"P-glycoproteins of Haemonchus contortus: development of real-time PCR assays for gene expression studies","volume":"86","author":"SM Williamson","year":"2012","journal-title":"J Helminthol"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0237150","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,8,6]],"date-time":"2020-08-06T15:02:54Z","timestamp":1596726174000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0237150"}},"subtitle":[],"editor":[{"given":"Adler R.","family":"Dillman","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2020,8,6]]},"references-count":41,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,8,6]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0237150","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,6]]}}}