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Juvenile oysters (Crassostrea virginica) respond to blue crab (Callinectus sapidus) chemical cues by growing stronger shells, while mud crabs (Panopeus herbstii) reduce foraging to avoid detection. However, the chemical composition of marine fear cues is largely unknown, partly due to methodological challenges. In this study, metabolomics was used to pinpoint predator metabolites that correlated with induced juvenile oyster defenses when exposed to the urine of blue crabs fed either oysters or mud crabs and to determine the chemical composition of these fear cues. Genetic algorithms applied to urine mass spectra coupled with bioassay data confirmed that metabolites from blue crab urine are responsible for fear responses by oysters. Sixteen compounds (of over 700) were determined to be potentially important cues used by juvenile oysters to detect blue crabs, with 3-(1-methyl-1H-imidazol-4-yl) propanoic acid identified as the most important contributor to this effect. These results indicate that oysters rely on certain metabolites that are ubiquitous in a variety of chemical blends to evaluate predation risk, and the response of juvenile oysters is not dependent on a singular metabolite but on numerous metabolites working in conjunction.<\/jats:p>","DOI":"10.1021\/acs.jproteome.6c00084","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T05:11:28Z","timestamp":1788325888000},"page":"5549-5561","source":"Crossref","is-referenced-by-count":0,"title":["Predator Cues\nInduce Defensive Responses in Marine\nPrey: A Metabolomics Decoding of Blue Crab Urine Biomarkers"],"prefix":"10.1021","volume":"25","author":[{"given":"Marisa R.","family":"Cepeda","sequence":"first","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benton R.","family":"Jaco","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin A.","family":"Belgrad","sequence":"additional","affiliation":[{"name":"Dauphin Island Sea Lab , , ,","place":["Dauphin Island, Alabama, United States, 36528"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria E.","family":"Branch","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel G.","family":"Moore","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah H.","family":"Roney","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9308-1895","authenticated-orcid":true,"given":"David A.","family":"Gaul","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Delbert L.","family":"Smee","sequence":"additional","affiliation":[{"name":"Dauphin Island Sea Lab , , ,","place":["Dauphin Island, Alabama, United States, 36528"]},{"name":"University of South Alabama , , , ,","place":["Mobile, Alabama, United States, 36608"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc J.","family":"Weissburg","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4482-1831","authenticated-orcid":true,"given":"Julia","family":"Kubanek","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,28]]},"reference":[{"key":"2026100208412589800_cit1","doi-asserted-by":"publisher","first-page":"199","DOI":"10.3354\/meps307199","article-title":"Fishes Learn\nAversions to a Nudibranchs Chemical Defense","volume":"307","author":"Long","year":"2006","journal-title":"Mar. Ecol.: Prog. Ser."},{"issue":"7","key":"2026100208412589800_cit2","doi-asserted-by":"publisher","first-page":"698","DOI":"10.1139\/Z10-029","article-title":"Chemical Ecology of Predator\u2013Prey\nInteractions in Aquatic Ecosystems: A Review and Prospectus","volume":"88","author":"Ferrari","year":"2010","journal-title":"Can. J. Zool."},{"issue":"11","key":"2026100208412589800_cit3","doi-asserted-by":"publisher","first-page":"2821","DOI":"10.1890\/0012-9658(2006)87[2821:CMCBMA]2.0.CO;2","article-title":"Chemically\nMediated Competition Between\nMicrobes and Animals: Microbes as Consumers in Food Webs","volume":"87","author":"Burkepile","year":"2006","journal-title":"Ecology"},{"issue":"6","key":"2026100208412589800_cit4","doi-asserted-by":"publisher","first-page":"1691","DOI":"10.2307\/1940703","article-title":"Predator-Induced\nMorphological Defenses: Costs, Life History Shifts, and Maternal Effects\nin Daphnia Pulex","volume":"76","author":"Tollrian","year":"1995","journal-title":"Ecology"},{"issue":"1","key":"2026100208412589800_cit5","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1007\/s00216-009-3162-5","article-title":"Searching for Signals\nin the Noise: Metabolomics in\nChemical Ecology","volume":"396","author":"Prince","year":"2010","journal-title":"Anal. Bioanal. Chem."},{"issue":"7","key":"2026100208412589800_cit6","doi-asserted-by":"publisher","first-page":"9181","DOI":"10.3390\/s120709181","article-title":"A Unifying\nReview of Bioassay-Guided Fractionation, Effect-Directed Analysis\nand Related Techniques","volume":"12","author":"Weller","year":"2012","journal-title":"Sensors"},{"issue":"7","key":"2026100208412589800_cit7","doi-asserted-by":"publisher","first-page":"937","DOI":"10.1039\/C5NP00003C","article-title":"Metabolomics in Chemical\nEcology","volume":"32","author":"Kuhlisch","year":"2015","journal-title":"Nat. Prod. Rep."},{"issue":"1","key":"2026100208412589800_cit8","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1007\/s00216-018-1470-3","article-title":"Simplifying the Complex:\nMetabolomics Approaches in\nChemical Ecology","volume":"411","author":"Poulin","year":"2019","journal-title":"Anal. Bioanal. 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Biofuels"},{"issue":"15","key":"2026100208412589800_cit12","doi-asserted-by":"publisher","first-page":"13899","DOI":"10.1021\/acsomega.3c00301","article-title":"Cryptic Chemical\nVariation in a Marine Red Alga as Revealed by Nontargeted Metabolomics","volume":"8","author":"Khatri Chhetri","year":"2023","journal-title":"ACS Omega"},{"issue":"3","key":"2026100208412589800_cit13","doi-asserted-by":"publisher","first-page":"36","DOI":"10.3390\/metabo7030036","article-title":"The Effect\nof Season on the Metabolic Profile of the European Clam Ruditapes\nDecussatus as Studied by 1H-NMR Spectroscopy","volume":"7","author":"Aru","year":"2017","journal-title":"Metabolites"},{"issue":"3","key":"2026100208412589800_cit14","doi-asserted-by":"publisher","first-page":"33","DOI":"10.3390\/metabo7030033","article-title":"NMR Profiling of Metabolites in Larval\nand Juvenile Blue Mussels (Mytilus Edulis) under Ambient and Low Salinity\nConditions","volume":"7","author":"May","year":"2017","journal-title":"Metabolites"},{"issue":"3","key":"2026100208412589800_cit15","doi-asserted-by":"publisher","first-page":"854","DOI":"10.1002\/anie.201208175","article-title":"Metabolomics Enables\nthe Structure Elucidation of a\nDiatom Sex Pheromone","volume":"52","author":"Gillard","year":"2013","journal-title":"Angew. 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