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We investigated the association between telework frequency and presence of non-COVID-19-related physical symptoms during the COVID-19 emergency declaration among workers in Japan. Data were collected from 1648 workers via web-based self-reported questionnaires. Multivariate logistic regression models were used to investigate the association between telework frequency per week (0, 1-2, 3-4, and ≥5 days/week) and the presence of stiff shoulders, eyestrain, and low back pain. Among 917 participants, telework frequency was significantly as