{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T06:33:34Z","timestamp":1793082814544,"version":"4.5.0"},"reference-count":25,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T00:00:00Z","timestamp":1792540800000},"content-version":"vor","delay-in-days":20,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001503","name":"Indian Council of Agricultural Research","doi-asserted-by":"publisher","award":["AINP\u2010LPPS"],"award-info":[{"award-number":["AINP\u2010LPPS"]}],"id":[{"id":"10.13039\/501100001503","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["ift.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Food Science"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:label\/>\n                    <jats:p>\n                      In this study, a spore\u2010based paper\u2010strip assay was developed for rapid, on\u2010site screening of antibiotics in milk. An \u03b1\u2010\n                      <jats:sc>d<\/jats:sc>\n                      \u2010glucosidase was identified as the optimal marker enzyme, producing a visible color signal within 30\u00a0min. Standardized production of\n                      <jats:italic>Geobacillus stearothermophilus<\/jats:italic>\n                      331 spores served as a stable biological sensing element. Optimization of germinants, nutrient supplements, sporulation conditions, and strip\u2010loading parameters enabled efficient spore germination and enzyme release, reducing assay time to 60\u201390\u00a0min. Thermal activation of spores at 100\u00b0C for 20\u00a0min improved sensitivity while maintaining signal clarity, and the final device was assembled on Whatman Grade 3 filter paper with immobilized chromogenic substrate and activated spores. The assay detected 42 antibiotics across 14 classes, including \u03b2\u2010lactams, aminoglycosides, tetracyclines, macrolides, sulfonamides, fluoroquinolones, polymyxins, nitrofurans, glycopeptides, ionophores, rifamycins, lincosamides, trimethoprims, and others, with limits of detection at or below regulatory maximum residue limits (MRLs). Field validation on 529 raw milk samples showed 15.63% positivity rate with 100% agreement with the commercial spore\u2010based kit and CHARM II and ROSA assays. The assay functions as a qualitative to semi\u2010quantitative screening tool based on visual color inhibition (where complete or partial lack of color development indicates residue levels at or near regulatory MRLs) and is not intended for exact quantification or identification of individual antibiotic concentrations, which requires instrument\u2010based confirmatory analytical methods. The assay also performed reliably on pasteurized and dried milk products. The developed strip assay offers a rapid, cost\u2010effective, user\u2010friendly, field\u2010deployable tool for routine screening of antibiotic residues in milk.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Practical Applications<\/jats:title>\n                    <jats:p>The developed spore\u2010based paper\u2010strip assay provides a rapid, cost\u2010effective, and portable tool for on\u2010site screening of antibiotic residues in milk without requiring sophisticated laboratory equipment. Its ability to detect 42 antibiotics from 14 classes within 60\u201390\u00a0min makes it suitable for routine monitoring at dairy farms, milk collection centers, and processing plants. The assay can facilitate an early identification of contaminated milk, thereby enhancing food safety, regulatory compliance, and antimicrobial stewardship across the dairy value chain.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/1750-3841.71564","type":"journal-article","created":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T04:33:42Z","timestamp":1792643622000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Rapid Spore\u2010Based Paper\u2010Strip Assay for Detection of Antibiotic Residues in Milk"],"prefix":"10.1111","volume":"91","author":[{"given":"Prashant","family":"Goel","sequence":"first","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Swathi P.","family":"Anand","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ravi","family":"Shukla","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alia","family":"Khan","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rushikesh","family":"Deshmukh","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naresh","family":"Kumar","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raghu Hirikyathanahalli","family":"Vishweswaraiah","sequence":"additional","affiliation":[{"name":"Microbial Biosensor, Food Safety and Quality Assurance Lab, Dairy Microbiology Division ICAR\u2014National Dairy Research Institute  Karnal Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,21]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365\u20102672.1998.00607.x"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1134\/S0026261709010081"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.2042-7158.1965.tb07734.x"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-96999-w"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13197\u2010011\u20100609\u20104"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2004.05.062"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010022\u201026466\u20107"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/1471-0307.13145"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102672.1994.tb04354.x"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.88.2.313\u2010318.1964"},{"key":"e_1_2_11_12_1","unstructured":"Kumar N. S.Das andM.Gaare.2016.Spore Germination\u2010Based Detection Kit for \u03b2\u2010Lactam Group in Milk. Indian Patent No. 273160.Indian Patent Office."},{"key":"e_1_2_11_13_1","unstructured":"Kumar N. A.Khan S.Arora et\u00a0al.2014.Development of Enzyme\u2010Spore\u2010BasedAssay(s) for Monitoring Antibiotic Residues in Milk. Indian Patent No. 365074.Indian Patent Office."},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11274\u2010012\u20101065\u20107"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10091124"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0734-9750(96)84850-6"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.crfs.2022.100416"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.5455\/javar.2019.f350"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01455-13"},{"key":"e_1_2_11_20_1","unstructured":"Shukla R.2016. \u201cSpore\u2010Based Paper Strip Assay for Detection of Antibiotic Residue in Milk.\u201d master's thesis ICAR\u2013National Dairy Research Institute."},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/jfs.12221"},{"key":"e_1_2_11_22_1","unstructured":"Shukla R.2016.Spore based paper strip assay for detection of antibiotic residues in milk (Master's thesis). Dairy Microbiology Division ICAR\u2010National Dairy Research Institute Karnal Haryana India."},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10311-018-00834-0"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaw1944"},{"key":"e_1_2_11_25_1","volume-title":"WHO Guidelines on Use of Medically Important Antimicrobials in Food\u2010Producing Animals","author":"World Health Organization","year":"2017"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1364\/OE.17.016480"}],"container-title":["Journal of Food Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/ift.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1750-3841.71564","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/ift.onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/1750-3841.71564","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/ift.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1750-3841.71564","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T05:46:58Z","timestamp":1793080018000},"score":1,"resource":{"primary":{"URL":"https:\/\/ift.onlinelibrary.wiley.com\/doi\/10.1111\/1750-3841.71564"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10]]},"references-count":25,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2026,10]]}},"alternative-id":["10.1111\/1750-3841.71564"],"URL":"https:\/\/doi.org\/10.1111\/1750-3841.71564","archive":["Portico"],"relation":{},"ISSN":["0022-1147","1750-3841"],"issn-type":[{"value":"0022-1147","type":"print"},{"value":"1750-3841","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10]]},"assertion":[{"value":"2026-06-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e71564"}}