{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T17:29:25Z","timestamp":1775237365030,"version":"3.50.1"},"reference-count":45,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,12,1]],"date-time":"2019-12-01T00:00:00Z","timestamp":1575158400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2019,12,19]],"date-time":"2019-12-19T00:00:00Z","timestamp":1576713600000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"published-print":{"date-parts":[[2019,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p>Although more modern methods are available, quantitative PCR (qPCR) is reproducible, sensitive and specific with instruments and expertise readily available in many laboratories. As such, the use of qPCR in <jats:italic>Cryptosporidium<\/jats:italic> research is well established and still widely used by researchers globally. This method depends upon the generation of standards at different concentrations to generate standard curves subsequently used for the quantification of DNA.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>We assessed four types of DNA template used to generate standard curves in drug screening studies involving <jats:italic>Cryptosporidium<\/jats:italic> spp.: (i) serially diluted <jats:italic>Cryptosporidium parvum<\/jats:italic> oocysts (10<jats:sup>6<\/jats:sup>\u20131); (ii) diluted template DNA from pure oocysts (\u00d710\u2013\u00d710<jats:sup>6<\/jats:sup> dilution of 10<jats:sup>6<\/jats:sup> oocyst DNA template); (iii) oocysts incubated in human ileocecal adenocarcinoma (HCT-8) cells (10<jats:sup>5<\/jats:sup>\u20131 and 5\u00a0\u00d7\u00a010<jats:sup>4<\/jats:sup>\u201350); and (iv) diluted DNA template (5\u00a0\u00d7\u00a010<jats:sup>4<\/jats:sup>) from cell culture incubated parasites (\u00d710\u2013\u00d71000).<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Serial dilutions of both cell culture and pure oocyst suspension DNA template yielded better linearity than cell culture derived standards, with dilutions of 10<jats:sup>6<\/jats:sup> oocysts exhibiting similar quantification cycle (Cq) values to those obtained from DNA template dilutions of 10<jats:sup>6<\/jats:sup> oocysts. In contrast, cell culture incubated oocysts demonstrated significantly higher DNA content than equivalent freely suspended oocysts and diluted DNA template from both cell culture derived and freely suspended oocysts across numerous concentrations.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>For many studies involving <jats:italic>Cryptosporidium<\/jats:italic>, only relative DNA content is required and as such, the superior linearity afforded by freely suspended oocysts and diluted DNA template (from either cell culture derived standards or freely suspended oocysts) will allow for more accurate relative quantification in each assay. Parasite division in the cell culture standards likely explains the higher DNA content found. These standards, therefore, have the potential to more accurately reflect DNA content in cell culture assays, and despite more modern methods available for absolute quantification, i.e. droplet digital PCR (ddPCR), the ubiquity of qPCR for the foreseeable future encourages further investigation into the reduced linearity observed in these standards such as varying oocyst seeding density, non-linear growth rates and assay efficiency.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s13071-019-3851-7","type":"journal-article","created":{"date-parts":[[2019,12,19]],"date-time":"2019-12-19T18:03:25Z","timestamp":1576778605000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Assessment of differences between DNA content of cell-cultured and freely suspended oocysts of Cryptosporidium parvum and their suitability as DNA standards in qPCR"],"prefix":"10.1186","volume":"12","author":[{"given":"Ian D.","family":"Woolsey","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Berit","family":"Blomstrand","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00d8ivind","family":"\u00d8ines","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heidi L.","family":"Enemark","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,12,19]]},"reference":[{"key":"3851_CR1","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1186\/s13567-017-0447-0","volume":"48","author":"S Thomson","year":"2017","unstructured":"Thomson S, Hamilton C, Hope J, Katzer F, Mabbott N, Morrison L, et al. Bovine cryptosporidiosis: impact, host-parasite interaction and control strategies. Vet Res. 2017;48:42.","journal-title":"Vet Res."},{"key":"3851_CR2","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1038\/503189a","volume":"503","author":"B Striepen","year":"2013","unstructured":"Striepen B. Parasitic infections: time to tackle cryptosporidiosis. Nature. 2013;503:189\u201391.","journal-title":"Nature."},{"key":"3851_CR3","first-page":"17","volume":"23","author":"M Bouwknegt","year":"2016","unstructured":"Bouwknegt M, Devleesschauwer B, Graham H, Robertson L, van der Giessen J. Prioritisation of food-borne parasites in Europe, 2016. Euro Surveill. 2016;23:17\u201300161.","journal-title":"Euro Surveill."},{"key":"3851_CR4","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.vetpar.2004.09.004","volume":"126","author":"R Fayer","year":"2004","unstructured":"Fayer R. Cryptosporidium: a water-borne zoonotic parasite. Vet Parasitol. 2004;126:37\u201356.","journal-title":"Vet Parasitol."},{"key":"3851_CR5","unstructured":"Casemore D, Watkins J. Review of disinfection and associated studies on Cryptosporidium. 1998. http:\/\/dwi.defra.gov.uk\/research\/completed-research\/reports\/dwi0805.pdf. Accessed 10 Oct 2018."},{"key":"3851_CR6","doi-asserted-by":"publisher","first-page":"1487","DOI":"10.1111\/j.1365-2672.2010.04764.x","volume":"109","author":"R Chalmers","year":"2010","unstructured":"Chalmers R, Giles M. Zoonotic cryptosporidiosis in the UK\u2014challenges for control. J Appl Microbiol. 2010;109:1487\u201397.","journal-title":"J Appl Microbiol."},{"key":"3851_CR7","doi-asserted-by":"publisher","first-page":"363","DOI":"10.17236\/sat00163","volume":"160","author":"P Olias","year":"2018","unstructured":"Olias P, Dettwiler I, Hemphill A, Deplazes P, Steiner A, Meylan M. The significance of cryptosporidiosis for the health of calves in Switzerland. Schweiz Arch Tierhelkd. 2018;160:363\u201374.","journal-title":"Schweiz Arch Tierhelkd."},{"key":"3851_CR8","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1111\/j.1574-6968.1999.tb08681.x","volume":"178","author":"A Armson","year":"1999","unstructured":"Armson A, Meloni P, Reynoldson J, Thompson R. Assessment of drugs against Cryptosporidium parvum using a simple in vitro screening method. FEMS Microbiol Lett. 1999;178:227\u201333.","journal-title":"FEMS Microbiol Lett."},{"key":"3851_CR9","doi-asserted-by":"publisher","first-page":"3809","DOI":"10.1128\/AEM.68.8.3809-3817.2002","volume":"68","author":"P Rochelle","year":"2002","unstructured":"Rochelle P, Marshall M, Mead J, Johnson A, Korich D, Rosen J, et al. Comparison of in vitro cell culture and a mouse assay for measuring infectivity of Cryptosporidium parvum. J Appl Environ Microbiol. 2002;68:3809\u201317.","journal-title":"J Appl Environ Microbiol."},{"key":"3851_CR10","doi-asserted-by":"publisher","first-page":"1155","DOI":"10.1007\/s00436-009-1535-5","volume":"105","author":"M Shahiduzzaman","year":"2009","unstructured":"Shahiduzzaman M, Dyachenko V, Khalafalla R, Desouky A, Daugschies A. Effects of curcumin on Cryptosporidium parvum in vitro. Parasitol Res. 2009;105:1155\u201361.","journal-title":"Parasitol Res."},{"key":"3851_CR11","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1007\/s00436-012-2824-y","volume":"111","author":"K Teichmann","year":"2012","unstructured":"Teichmann K, Kuliberda M, Schatzmayr G, Hadacek F, Joachim A. In vitro determination of anticryptosporidial activity of phytogenic extracts and compounds. Parasitol Res. 2012;111:231\u201340.","journal-title":"Parasitol Res."},{"key":"3851_CR12","doi-asserted-by":"publisher","first-page":"170","DOI":"10.1016\/j.parint.2017.11.001","volume":"67","author":"S Gaur","year":"2018","unstructured":"Gaur S, Kuhlenschmidt T, Kuhlenschmidt M, Andrade J. Effect of oregano essential oil and carvacrol on Cryptosporidium parvum infectivity in HCT-8 cells. Parasitol Int. 2018;67:170\u20135.","journal-title":"Parasitol Int."},{"key":"3851_CR13","doi-asserted-by":"publisher","first-page":"1048","DOI":"10.1016\/S0020-7519(01)00212-0","volume":"31","author":"N Hijjawi","year":"2001","unstructured":"Hijjawi N, Meloni B, Ryan U, Thompson R. Complete development and long-term maintenance of Cryptosporidium parvum human and cattle genotypes in cell culture. Int J Parasitol. 2001;31:1048\u201355.","journal-title":"Int J Parasitol."},{"key":"3851_CR14","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1007\/s00436-006-0437-z","volume":"101","author":"M Najdrowski","year":"2007","unstructured":"Najdrowski M, Heckeroth A, Wackwitz C, Gawlowska S, Mackenstedt U, Kliemt D, et al. Development and validation of a cell culture based assay for in virto assessment of anticryptosporidial compounds. Parasitol Res. 2007;101:161\u20137.","journal-title":"Parasitol Res."},{"key":"3851_CR15","doi-asserted-by":"publisher","first-page":"1495","DOI":"10.1128\/AEM.71.3.1495-1500.2005","volume":"71","author":"G Giovanni","year":"2005","unstructured":"Giovanni G, LeChevallier M. Quantitative-PCR assessment of Cryptosporidium parvum cell culture infection. Appl Environ Microbiol. 2005;71:1495\u2013500.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR16","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1017\/S0031182011000217","volume":"138","author":"B King","year":"2011","unstructured":"King B, Keegan A, Robinson B, Monis P. Cryptosporidium cell culture infectivity assay design. Parasitology. 2011;138:671\u201381.","journal-title":"Parasitology."},{"key":"3851_CR17","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1126\/science.6710159","volume":"224","author":"W Current","year":"1984","unstructured":"Current W, Haynes T. Complete development of Cryptosporidium in cell cultures. Science. 1984;224:603\u20135.","journal-title":"Science."},{"key":"3851_CR18","doi-asserted-by":"publisher","first-page":"855","DOI":"10.1017\/S0031182017001937","volume":"145","author":"P Karanis","year":"2018","unstructured":"Karanis P. The truth about in vitro culture of Cryptosporidium species. Parasitology. 2018;145:855\u201364.","journal-title":"Parasitology."},{"key":"3851_CR19","doi-asserted-by":"publisher","first-page":"8604","DOI":"10.1021\/ac202028g","volume":"83","author":"B Hindsom","year":"2011","unstructured":"Hindsom B, Ness K, Masquelier D, Belgrader P, Heredia N, Makarewicz A, et al. High-throughput droplet digital PCR system for absolute quantification of DNA copy number. Anal Chem. 2011;83:8604\u201310.","journal-title":"Anal Chem."},{"key":"3851_CR20","doi-asserted-by":"publisher","first-page":"4484","DOI":"10.1128\/AEM.00189-06","volume":"72","author":"N Godiwala","year":"2006","unstructured":"Godiwala N, Vandewalle A, Ward H, Leav B. Quantification of in vitro and in vivo Cryptosporidium parvum infection by using real-time PCR. Appl Environ Microbiol. 2006;72:4484\u20138.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR21","doi-asserted-by":"publisher","first-page":"1105","DOI":"10.1016\/j.ijpara.2014.08.004","volume":"44","author":"R Yang","year":"2014","unstructured":"Yang R, Paparini A, Monis P, Ryan U. Comparison of next-generation droplet digital PCR (ddPCR) with quantitative PCR (qPCR) for enumeration of Crptosporidium oocysts in faecal samples. Int J Parasitol. 2014;44:1105\u201313.","journal-title":"Int J Parasitol."},{"key":"3851_CR22","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.vetpar.2009.12.021","volume":"169","author":"N Hijjawi","year":"2010","unstructured":"Hijjawi N, Estcourt A, Yang R, Monis P, Ryan U. Complete development and multiplication of Cryptosporidium hominis in cell-free culture. Vet Parasitol. 2010;169:29\u201336.","journal-title":"Vet Parasitol."},{"key":"3851_CR23","doi-asserted-by":"publisher","first-page":"2505","DOI":"10.1128\/AEM.69.5.2505-2511.2003","volume":"69","author":"A Keegan","year":"2003","unstructured":"Keegan A, Fanok S, Monis P, Saint C. Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection. Appl Environ Microbiol. 2003;69:2505\u201311.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR24","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1016\/j.mimet.2010.01.017","volume":"80","author":"M Garvey","year":"2010","unstructured":"Garvey M, Farrell H, Cormican M, Rowan N. Investigations of the relationship between the use of in vitro cell culture-quantitative PCR and a mouse-based bioassay for evaluating critical factors affecting the disinfection performance of pulsed UV light for treating Cryptosporidium parvum oocysts in saline. J Microbiol Meth. 2010;80:267\u201373.","journal-title":"J Microbiol Meth."},{"key":"3851_CR25","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1016\/j.mimet.2012.11.015","volume":"92","author":"E Koken","year":"2013","unstructured":"Koken E, Darnault J, Jacobson A, Powelson D, Hendrickson W. Quantification of Cryptosporidium parvum in natural soil matrices and soil solutions using qPCR. J Microbial Meth. 2013;92:135\u201344.","journal-title":"J Microbial Meth."},{"key":"3851_CR26","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1016\/S0166-6851(02)00041-5","volume":"121","author":"L MacDonald","year":"2002","unstructured":"MacDonald L, Sargent K, Armson A, Thompson A, Reynoldson J. The development of a real-time quantitative-PCR method for characterisation of a Cryptosporidium parvum in vitro culturing system and assessment of drug efficacy. Mol Biochem Parasitol. 2002;121:279\u201382.","journal-title":"Mol Biochem Parasitol."},{"key":"3851_CR27","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1016\/j.vetpar.2009.03.009","volume":"162","author":"M Shahiduzzaman","year":"2009","unstructured":"Shahiduzzaman M, Dyachenko V, Obwaller A, Unglaube S, Daugschies A. Combination of cell culture and quantitative PCR for screening of drugs against Cryptosporidium parvum. Vet Parasitol. 2009;162:271\u20137.","journal-title":"Vet Parasitol."},{"key":"3851_CR28","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/j.vetpar.2009.09.042","volume":"164","author":"M Shahiduzzaman","year":"2010","unstructured":"Shahiduzzaman M, Dyachenko V, Keidel J, Sch\u00e4schke R, Daugschies A. Combination of cell culture and quantitative PCR (cc-qPCR) to assess disinfectants efficacy on Cryptosporidium oocysts under standardized conditions. Vet Parasitol. 2010;164:43\u20139.","journal-title":"Vet Parasitol."},{"key":"3851_CR29","doi-asserted-by":"publisher","first-page":"e51232","DOI":"10.1371\/journal.pone.0051232","volume":"7","author":"S Benamrouz","year":"2012","unstructured":"Benamrouz S, Guyot K, Gazzola S, Mouray A, Chassat T, Delaire B, et al. Cryptosporidium parvum infection in SCID mice infected with only one oocyst: qPCR assessment of parasite replication in tissues and development of digestive cancer. PLoS ONE. 2012;7:e51232.","journal-title":"PLoS ONE"},{"key":"3851_CR30","first-page":"1","volume":"25","author":"G Leich","year":"2011","unstructured":"Leich G, He Q. Cryptosporidiosis\u2014an overview. J Biomed Sci. 2011;25:1\u201316.","journal-title":"J Biomed Sci."},{"key":"3851_CR31","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1017\/S0031182009990837","volume":"137","author":"H Borowski","year":"2010","unstructured":"Borowski H, Thompson R, Armstrong T, Clode P. Morphological characterization of Cryptosporidium parvum life-cycle stages in an in vitro model system. Parasitology. 2010;137:13\u201326.","journal-title":"Parasitology."},{"key":"3851_CR32","doi-asserted-by":"publisher","first-page":"e93","DOI":"10.1093\/nar\/gng093","volume":"31","author":"R Rutledge","year":"2003","unstructured":"Rutledge R, C\u00f4te C. Mathematics of quantitative kinetic PCR and the application of standard curves. Nucleic Acids Res. 2003;31:e93.","journal-title":"Nucleic Acids Res."},{"key":"3851_CR33","doi-asserted-by":"crossref","first-page":"3488","DOI":"10.1128\/AEM.58.11.3488-3493.1992","volume":"58","author":"A Campbell","year":"1992","unstructured":"Campbell A, Robertson L, Smith H. Viability of Cryptosporidium parvum oocysts: correlation of in vitro excystation with inclusion or exclusion of fluorogenic vital dyes. Appl Environ Microbiol. 1992;58:3488\u201393.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR34","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1089\/fpd.2017.2348","volume":"15","author":"H Petersen","year":"2018","unstructured":"Petersen H, Enemark H. Viability assessment of Cryptosporidium oocysts by vital dyes: dry mounts overestimate the number of \u201cghost\u201d oocysts. Foodborne Pathog Dis. 2018;15:141\u20134.","journal-title":"Foodborne Pathog Dis."},{"key":"3851_CR35","doi-asserted-by":"crossref","first-page":"3669","DOI":"10.1128\/AEM.63.9.3669-3675.1997","volume":"63","author":"T Slifko","year":"1997","unstructured":"Slifko T, Friedman D, Rose J, Jakubowski W. An in vitro method for detecting infectious Cryptosporidium oocysts with cell culture. Appl Environ Microbiol. 1997;63:3669\u201375.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR36","doi-asserted-by":"publisher","first-page":"825","DOI":"10.2307\/3284275","volume":"83","author":"U Morgan","year":"1997","unstructured":"Morgan U, Constantine C, Forbes D, Thompson A. Differentiation between human and animal isolates of Cryptosporidium parvum using rDNA sequencing and direct PCR analysis. J Parasitol. 1997;83:825\u201330.","journal-title":"J Parasitol."},{"key":"3851_CR37","doi-asserted-by":"crossref","first-page":"1919","DOI":"10.1128\/AEM.56.6.1919-1925.1990","volume":"56","author":"R Amman","year":"1990","unstructured":"Amman R, Binder B, Olsen R, Chislom R, Devereaux R, Stahl D. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analysing mixed microbial populations. Appl Environ Microbiol. 1990;56:1919\u201325.","journal-title":"Appl Environ Microbiol."},{"key":"3851_CR38","unstructured":"BioRad. CFX Manager Version 31. Hercules: BioRad; 2019."},{"key":"3851_CR39","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","volume":"55","author":"S Bustin","year":"2009","unstructured":"Bustin S, Benes V, Garson J, Hellemans J, Huggestt J, Kubista M, Mueller R, et al. The MIQE Guidelines: minimum information for publication of quantitative real-time PCR Experiments. Clin Chem. 2009;55:611\u201322.","journal-title":"Clin Chem."},{"key":"3851_CR40","unstructured":"R Core Team (2019). R: a language and environment for statistical computing. Vienna: R Foundation for Statistical Computing. http:\/\/www.r-project.org\/."},{"key":"3851_CR41","doi-asserted-by":"publisher","first-page":"986","DOI":"10.1101\/gr.6.10.986","volume":"6","author":"C Heid","year":"1996","unstructured":"Heid C, Stevens J, Livak K, Williams P. Real time quantitative PCR. Genome Res. 1996;6:986\u201394.","journal-title":"Genome Res."},{"key":"3851_CR42","doi-asserted-by":"publisher","first-page":"2556","DOI":"10.1128\/JCM.03458-12","volume":"51","author":"C Mary","year":"2013","unstructured":"Mary C, Chapey E, Dutoit K, Guyot K, Hasseine L, Jeddl F, et al. Multicentric evaluation of a new real-time PCR assay for quantification of Cryptosporidium spp. and identification of Cryptosporidium parvum and Cryptosporidium hominis. J Clin Microbiol. 2013;51:2556\u201363.","journal-title":"J Clin Microbiol."},{"key":"3851_CR43","doi-asserted-by":"publisher","first-page":"142","DOI":"10.1016\/j.exppara.2013.06.014","volume":"135","author":"R Yang","year":"2013","unstructured":"Yang R, Murphy C, Song Y, Ng-Hublin J, Estcourt A, Hijjawi N, et al. Specific and quantitative detection and identification of Cryptosporidium hominis and C. parvum in clinical and environmental samples. Exp Parasitol. 2013;135:142\u20137.","journal-title":"Exp Parasitol."},{"key":"3851_CR44","doi-asserted-by":"publisher","first-page":"951","DOI":"10.1007\/s00436-007-0567-y","volume":"101","author":"A Leetz","year":"2007","unstructured":"Leetz A, Sotiriadou I, Ongerth J, Karanis P. An evaluation of primers amplifying DNA targets for the detection of Cryptosporidium spp. using C. parvum HNJ-1 Japanese isolate in water samples. Parasitol Res. 2007;101:951\u201362.","journal-title":"Parasitol Res."},{"key":"3851_CR45","unstructured":"Qiagen. QIAamp\u00ae DNA Mini and Blood Mini Handbook. Qiagen. 2016. https:\/\/www.qiagen.com\/ch\/resources\/download.aspx?id=62a200d6-faf4-469b-b50f-2b59cf738962&lang=en. Accessed 31 July 2019."}],"container-title":["Parasites &amp; Vectors"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-019-3851-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/s13071-019-3851-7\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-019-3851-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,12,18]],"date-time":"2020-12-18T00:52:34Z","timestamp":1608252754000},"score":1,"resource":{"primary":{"URL":"https:\/\/parasitesandvectors.biomedcentral.com\/articles\/10.1186\/s13071-019-3851-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12]]},"references-count":45,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2019,12]]}},"alternative-id":["3851"],"URL":"https:\/\/doi.org\/10.1186\/s13071-019-3851-7","relation":{},"ISSN":["1756-3305"],"issn-type":[{"value":"1756-3305","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12]]},"assertion":[{"value":"3 September 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 December 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 December 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Not applicable.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"596"}}