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While it has been scientifically proven that COVID-19 vaccine is a safe and effective measure to reduce the severity of infection and curbing the spread of the SARS-CoV-2 virus, skepticism remains widespread, and in many countries vaccine mandates have been met with strong opposition. In this study, we applied machine learning-based analyses of the U.S.-based tweets covering the periods leading toward and after the Biden Administration’s announcement of federal vaccine mandates, supplemented by a qualitative content analysis of a random sample of relevant tweets. The objective was to examine the beliefs held among twitter users toward vaccine mandates, as well as the evidence that they used to support their positions. The results show that while approximately 30% of the twitter users included in the dataset supported the measure, more users expressed differing opinions. Concerns raised included questioning on the political motive, infringement of personal liberties, and ineffectiveness in preventing infection.