{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T16:01:48Z","timestamp":1790438508109,"version":"4.1.0"},"reference-count":20,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Ixodid ticks are among the most widespread and permissive groups of arthropod disease vectors in the world. They transmit various viral, bacterial, protozoan, and even nematode pathogens infective for humans and animals. Ticks are essential to studying pathogen transmission and infection in a new host; however, exposing in vivo models to ticks is very challenging. Most current tick placement protocols rely on capsules to increase attachment rates at a specific location. However, capsules interfere with hosts\u2019 ability to engage with ticks by grooming\/scratching. Conversely, available capsule\u2010free tick infestation protocols, including free\u2010roaming protocols, suffer from low and highly variable tick attachment efficacy. Here, we describe a highly efficient capsule\u2010free nymphal tick placement protocol with tick attachment rates of \u226585% across various tick species and genera, offering the generation of highly reproducible data. This article also offers a set of protocols to the user primarily geared toward the study of itch\u2010induced tick removal but are adaptable to other study settings. Published 2026. This article is a U.S. Government work and is in the public domain in the USA. Current Protocols published by Wiley Periodicals LLC.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Capsule\u2010free, highly efficient multi\u2010species nymphal tick placement for site\u2010specific attachment\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Assessing efficacy of itch\u2010induced tick removal by quantifying loss of ticks over time\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Assessment of health\/ability of partially fed nymphal ticks to feed determined by scutal index\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 4<\/jats:bold>\n                    : Scratch bout quantification in guinea pigs\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : Development of a semi\u2010supervised deep learning pipeline for automated detection and validation of scratching behavior\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 5<\/jats:bold>\n                    : Assessment of local skin inflammation due to immunity to tick bites using laser speckle contrast imaging\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70477","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T15:29:26Z","timestamp":1790177366000},"source":"Crossref","is-referenced-by-count":0,"title":["Efficient, Capsule\u2010Free Nymphal Ixodid Tick Placement for Site\u2010Specific Attachment to Assess Anti\u2010Tick Host Behavior and Immune Responses"],"prefix":"10.1002","volume":"6","author":[{"given":"Ronja A.","family":"Frigard","sequence":"first","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serena","family":"Doh","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles S.","family":"Grugan","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanling","family":"Liu","sequence":"additional","affiliation":[{"name":"Integrated Data Sciences Section, Research Technologies Branch, Division of Intramural Research National Institute of Allergy and Infectious Diseases  Bethesda Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara I.","family":"Mirbagheri","sequence":"additional","affiliation":[{"name":"Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Bethesda Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luana A.","family":"Rogerio","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Iniguez","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiago D.","family":"Serafim","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline da Silva","family":"Moreira","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jack","family":"Collins","sequence":"additional","affiliation":[{"name":"Integrated Data Sciences Section, Research Technologies Branch, Division of Intramural Research National Institute of Allergy and Infectious Diseases  Bethesda Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel E.","family":"Sonenshine","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesus G.","family":"Valenzuela","sequence":"additional","affiliation":[{"name":"Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health  Rockville Maryland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes S. 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