{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T13:30:43Z","timestamp":1791984643288,"version":"4.4.0"},"reference-count":67,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T00:00:00Z","timestamp":1750982400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR\u201021\u2010CE35\u20100016"],"award-info":[{"award-number":["ANR\u201021\u2010CE35\u20100016"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ECOPATHS"],"award-info":[{"award-number":["ECOPATHS"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["TVACALBA"],"award-info":[{"award-number":["TVACALBA"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004796","name":"Institut Polaire Fran\u00e7ais Paul Emile Victor","doi-asserted-by":"publisher","award":["IPEV ECOPATH\u20101151"],"award-info":[{"award-number":["IPEV ECOPATH\u20101151"]}],"id":[{"id":"10.13039\/501100004796","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019184","name":"Biodiversa+","doi-asserted-by":"publisher","award":["ANR\u201021\u2010BIRE\u20100006"],"award-info":[{"award-number":["ANR\u201021\u2010BIRE\u20100006"]}],"id":[{"id":"10.13039\/100019184","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019184","name":"Biodiversa+","doi-asserted-by":"publisher","award":["REMOVE_DISEASE"],"award-info":[{"award-number":["REMOVE_DISEASE"]}],"id":[{"id":"10.13039\/100019184","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012681","name":"College of Natural Resources and Sciences, Humboldt State University","doi-asserted-by":"publisher","award":["Zone\u2010Atelier\u2010Antarctique\u2010et\u2010Terres\u2010Australes"],"award-info":[{"award-number":["Zone\u2010Atelier\u2010Antarctique\u2010et\u2010Terres\u2010Australes"]}],"id":[{"id":"10.13039\/100012681","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012681","name":"College of Natural Resources and Sciences, Humboldt State University","doi-asserted-by":"publisher","award":["SEE\u2010Life\u2010ECOPATH"],"award-info":[{"award-number":["SEE\u2010Life\u2010ECOPATH"]}],"id":[{"id":"10.13039\/100012681","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["besjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Animal Ecology"],"published-print":{"date-parts":[[2025,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>\n<jats:list>\n\n<jats:list-item><jats:p>Among recent advances in molecular biology for studying infectious diseases, the microfluidic high\u2010throughput real\u2010time polymerase chain reaction (Htrt PCR) has emerged as an efficient first\u2010line tool for the detection of a wide range of infectious agents (IA) in a host system. This technology allows large batches of samples to be screened simultaneously for tens of targeted IA by real\u2010time PCR. It represents a promising tool in disease ecology.<\/jats:p><\/jats:list-item>\n\n<jats:list-item><jats:p>As a proof of concept, we present here the development steps and initial application of a microfluidic Htrt PCR system for the detection of DNA from 28 selected IA in a set of wild vertebrates. We applied this approach to 497 samples (mainly mucosal swabs and necropsy tissues) from 274 seabirds and 80 mammals from Southern Ocean islands. This ecosystem is of particular interest for disease ecology and biodiversity conservation due to the high host densities of breeding colonies, within and among which pathogens may spread rapidly. Positive samples were subsequently confirmed for each PCR system using a secondary real\u2010time or conventional PCR system and\/or sequencing.<\/jats:p><\/jats:list-item>\n\n<jats:list-item><jats:p>Fourteen targeted IA were detected. The approach allowed an efficient screening of host species for known seabird pathogens, including <jats:italic>Pasteurella multocida<\/jats:italic> (avian cholera) (9.9% [6.6\u201314.0] in seabirds, 18.3% [9.5\u201330.4] in mammals) and <jats:italic>Erysipelothrix amsterdamensis<\/jats:italic> (15% [11\u201319.8] in seabirds, 2.1% [0.4\u20136.1] in mammals) in cloacal and rectal swabs. Their detection on islands where they were not known previously raised conservation concerns. IA not previously known to be circulating in the system were detected at high prevalence, notably <jats:italic>Chlamydiaceae<\/jats:italic> on all colonies (32.1% [26.6\u201338] in seabirds and 21.3% [12.9\u201331.8] in mammals, in cloacal and rectal swabs). Prevalence and diversity of targeted IA could be compared among sites and host species, highlighting the usefulness of the approach to explore drivers of IA community dynamics, but also to identify specific hosts as potential epidemiological sentinels or reservoirs.<\/jats:p><\/jats:list-item>\n\n<jats:list-item><jats:p>Htrt PCR is a powerful and versatile tool that can be used in disease ecology for exploring the composition of IA communities within host communities, but also for addressing other important basic and applied questions in multi\u2010host, multi\u2010pathogen systems.<\/jats:p><\/jats:list-item>\n<\/jats:list>\n<\/jats:p>","DOI":"10.1111\/1365-2656.70088","type":"journal-article","created":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T07:44:11Z","timestamp":1751010251000},"page":"1625-1637","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["High\u2010throughput microfluidic real\u2010time <scp>PCR<\/scp> as a promising tool in disease ecology"],"prefix":"10.1111","volume":"94","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-1377-849X","authenticated-orcid":false,"given":"Tristan","family":"Bralet","sequence":"first","affiliation":[{"name":"CEFE, CNRS, Universit\u00e9 Montpellier, EPHE, IRD  Montpellier France"},{"name":"Anses, Animal Health Laboratory, Bacterial Zoonoses Unit  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachid","family":"Aaziz","sequence":"additional","affiliation":[{"name":"Anses, Animal Health Laboratory, Bacterial Zoonoses Unit  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00e9r\u00e9my","family":"Tornos","sequence":"additional","affiliation":[{"name":"CEFE, CNRS, Universit\u00e9 Montpellier, EPHE, IRD  Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amandine","family":"Gamble","sequence":"additional","affiliation":[{"name":"School of Biodiversity, One Health and Veterinary Medicine University of Glasgow  Glasgow UK"},{"name":"Department of Public and Ecosystem Health Cornell University  Ithaca New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Augustin","family":"Clessin","sequence":"additional","affiliation":[{"name":"CEFE, CNRS, Universit\u00e9 Montpellier, EPHE, IRD  Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathilde","family":"Lejeune","sequence":"additional","affiliation":[{"name":"CEFE, CNRS, Universit\u00e9 Montpellier, EPHE, IRD  Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cl\u00e9mence","family":"Galon","sequence":"additional","affiliation":[{"name":"ANSES, INRAE, Ecole Nationale V\u00e9t\u00e9rinaire d'Alfort, UMR BIPAR, Laboratoire de Sant\u00e9 Animale  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorraine","family":"Michelet","sequence":"additional","affiliation":[{"name":"Anses, Animal Health Laboratory, Bacterial Zoonoses Unit  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C\u00e9lia","family":"Lesage","sequence":"additional","affiliation":[{"name":"Environmental Department Terres Australes et Antarctiques Fran\u00e7aises (TAAF)  Saint\u2010Pierre La R\u00e9union France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiphaine","family":"Jeanniard du Dot","sequence":"additional","affiliation":[{"name":"Centre d'Etudes Biologiques de Chiz\u00e9, Centre National de la Recherche Scientifique  Villiers\u2010en\u2010Bois France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillaume","family":"Desoubeaux","sequence":"additional","affiliation":[{"name":"Parasitology and Mycology Department Bretonneau Hospital  Tours France"},{"name":"Research Center for Respiratory Diseases Inserm UMR1100, University of Tours  Tours France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muriel","family":"Guyard","sequence":"additional","affiliation":[{"name":"Anses, Ploufragan\u2010Plouzan\u00e9 Laboratory Unit of Hygiene and Quality of Poultry and Pork Products  Ploufragan France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Delannoy","sequence":"additional","affiliation":[{"name":"Anses, Laboratory for Food Safety COLiPATH Unit &amp; Genomics Platform IdentyPath  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Moutailler","sequence":"additional","affiliation":[{"name":"ANSES, INRAE, Ecole Nationale V\u00e9t\u00e9rinaire d'Alfort, UMR BIPAR, Laboratoire de Sant\u00e9 Animale  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karine","family":"Laroucau","sequence":"additional","affiliation":[{"name":"Anses, Animal Health Laboratory, Bacterial Zoonoses Unit  Maisons\u2010Alfort France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thierry","family":"Boulinier","sequence":"additional","affiliation":[{"name":"CEFE, CNRS, Universit\u00e9 Montpellier, EPHE, IRD  Montpellier France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,6,27]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/ani12101226"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008135"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens12070937"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10344\u2010024\u201001799\u2010y"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.onehlt.2022.100479"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2018.2772"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1080\/00063657.2024.2396563"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/1755\u20100998.12946"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1579\/0044\u20107447\u201034.6.430"},{"key":"e_1_2_9_11_1","unstructured":"Boulinier T. Gamble A. Tornos J. &Garnier R.(2016).Circulation d'agents infectieux dans les populations de vert\u00e9br\u00e9s coloniaux des terres australes:Surveillance compr\u00e9hension et implications pour la gestion. Rapport d'activit\u00e9 2016\u20102017\u2014PROGRAMME ECOPATH 1151 (pp. 52\u201361). IPEV \u2022 Institut polaire fran\u00e7ais Paul\u2010\u00c9mile Victor.https:\/\/institut\u2010polaire.fr\/wp\u2010content\/uploads\/2021\/06\/WEB_124004_rapport_activite_2016.pdf"},{"key":"e_1_2_9_12_1","volume-title":"Behavioural ecology of lesser sheathbill (Chionis minor) at Marion Island","author":"Burger A. E.","year":"1980"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1303235"},{"key":"e_1_2_9_14_1","doi-asserted-by":"crossref","unstructured":"Clessin A. Briand F.\u2010X. Tornos J. Lejeune M. Pasquale C. D. Fischer R. Souchaud F. Hirchaud E. Bralet T. Guinet C. McMahon C. R. Grasland B. Baele G. &Boulinier T.(2025).Mass mortality events in the sub\u2010Antarctic Indian Ocean caused by long\u2010distance circumpolar spread of highly pathogenic avian influenza H5N1 clade 2.3.4.4b.https:\/\/doi.org\/10.1101\/2025.02.25.640068","DOI":"10.1101\/2025.02.25.640068"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.5038\/2074-1235.37.3.843"},{"key":"e_1_2_9_16_1","volume-title":"Molecular epidemiology: Principles and practices","author":"Coutl\u00e9e F.","year":"2011"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1080\/03079457.2018.1430891"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2023.110303"},{"key":"e_1_2_9_19_1","volume-title":"Atlas of top predators from French Southern Territories in the Southern Indian Ocean","author":"Delord K.","year":"2014"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biocon.2019.06.033"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abc3189"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010022\u201005506\u20102"},{"key":"e_1_2_9_23_1","volume-title":"Encyclopedia of ecology","author":"Fath B. D.","year":"2018"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/brv.12380"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00442\u2010019\u201004369\u20101"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1752\u20104571.2010.00127.x"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.3354\/meps08212"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00300\u2010014\u20101632\u20105"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/jwmg.22588"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12041\u2010020\u201001225\u20107"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/tmi.13965"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1098\/rsif.2007.0237"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/ece3.4013"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00300\u2010015\u20101748\u20102"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10393\u2010020\u201001487\u20108"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.7589\/JWD\u2010D\u201020\u201000177"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/ece3.11382"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1111\/ele.13875"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.amb.2019.02.001"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1111\/mec.16398"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac0346407"},{"key":"e_1_2_9_42_1","first-page":"1","article-title":"Use of \u201clysis buffer\u201d in DNA isolation and its implications for museum collections","volume":"163","author":"Longmire J.","year":"1997","journal-title":"Museum of Texas Tech University"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010024\u201007380\u20106"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1089\/vbz.2018.2277"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1002448"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1461-0248.2010.01472.x"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2014.00103"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.3390\/v11100904"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0178811"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.1002\/ece3.8916"},{"key":"e_1_2_9_51_1","doi-asserted-by":"crossref","unstructured":"Ostaizka A. Dunn R. R. Hansen L. H. Gilbert M. T. P. &Alberdi A.(2023).Field and laboratory guidelines for reliable bioinformatic and statistical analysis of bacterial shotgun metagenomic data 44(6).https:\/\/doi.org\/10.1080\/07388551.2023.2254933","DOI":"10.1080\/07388551.2023.2254933"},{"key":"e_1_2_9_52_1","volume-title":"R: A language and environment for statistical computing","author":"R Core Team","year":"2021"},{"key":"e_1_2_9_53_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0954102017000347"},{"key":"e_1_2_9_54_1","doi-asserted-by":"publisher","DOI":"10.1093\/biosci\/biw175"},{"key":"e_1_2_9_55_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.2041\u2010210X.2012.00215.x"},{"key":"e_1_2_9_56_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1472\u20104642.2006.00279.x"},{"key":"e_1_2_9_57_1","volume-title":"Veterinary immunology","author":"Tizard I. R.","year":"2013"},{"key":"e_1_2_9_58_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102656.2010.01742.x"},{"key":"e_1_2_9_59_1","doi-asserted-by":"crossref","unstructured":"Tornos J. Bralet T. &Boulinier T.(2025).High\u2010throughput microfluidic real\u2010time PCR as a promising tool in disease ecology [Dataset]. data.InDoRES.https:\/\/doi.org\/10.48579\/PRO\/HZCSWK","DOI":"10.1111\/1365-2656.70088"},{"key":"e_1_2_9_60_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0959270916000629"},{"key":"e_1_2_9_61_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-030-16501-7_7"},{"key":"e_1_2_9_62_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.syapm.2021.126200"},{"key":"e_1_2_9_63_1","doi-asserted-by":"publisher","DOI":"10.1002\/ecy.1675"},{"key":"e_1_2_9_64_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.OH\u20100001\u20102012"},{"key":"e_1_2_9_65_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008940"},{"key":"e_1_2_9_66_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0169\u20105347(01)02205\u20104"},{"key":"e_1_2_9_67_1","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.03302\u201022"},{"key":"e_1_2_9_68_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijsem.0.006264"}],"container-title":["Journal of Animal Ecology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/besjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1365-2656.70088","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T11:16:49Z","timestamp":1757589409000},"score":1,"resource":{"primary":{"URL":"https:\/\/besjournals.onlinelibrary.wiley.com\/doi\/10.1111\/1365-2656.70088"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,27]]},"references-count":67,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2025,9]]}},"alternative-id":["10.1111\/1365-2656.70088"],"URL":"https:\/\/doi.org\/10.1111\/1365-2656.70088","archive":["Portico"],"relation":{},"ISSN":["0021-8790","1365-2656"],"issn-type":[{"value":"0021-8790","type":"print"},{"value":"1365-2656","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,27]]},"assertion":[{"value":"2025-01-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-18","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-06-27","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}