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As the U.S. largest minority group, Hispanic population suffers from significant health disparities. Literature suggests that adequate access to credible health information is essential to achieve better health outcomes. However, few studies have systematically examined U.S. Hispanics’ health information seeking beliefs and behaviors. Based on bidimensional model of acculturation and comprehensive model of information seeking, our study investigated how U.S. Hispanics’ acculturation to U.S. and Hispanic cultures is related to their trust and seeking health information from Internet, interpersonal sources, and traditional media. We conducted structural equation modeling analysis of data from 2009-2012 Annenberg National Health Communication Surveys. Results suggested that U.S. Hispanics had different acculturation strategies, which were differentially associated with their trust and seeking health information from various information sources. Trust in an information source mediated the relationship between acculturation and health information seeking from the source. Theoretical and practical implications of our research were discussed.