Context: The COVID-19 pandemic and other public health challenges have increased the need for longitudinal data quantifying the changes in the state public health workforce. Objective: To characterize the state of governmental public health workforce among state health agency (SHA) staff across the United States and provide longitudinal comparisons to 2 prior fieldings of the survey. Design: State health agency leaders were invited to have their workforce to participate in PH WINS 2021. As in prior fieldings, participating agencies provided staff lists used to send e-mail invitations to employees to participate in this electronic survey. Setting and Participants: State health agency staff. Main Outcome Measures: PH WINS 2021 maintains the 4 primary domains from 2014 and 2017 (ie, workplace engagement, training needs assessment, emerging public health concepts, and demographics) and includes new questions related to the mental and emotional well-being; the impact of the COVID-19 pandemic on staff retention; and the workforce's awareness of and confidence in emerging public health concepts. Results: The percentage of SHA staff who self-identify as Black, Indigenous, and people of color increased from 30% (95% confidence interval [CI]: 29%-32%) to 35% (95% CI: 35%-37%) between 2014 and 2021. Staff younger than 31 years accounted for 11% (95% CI: 10%-12%) of the SHA workforce in 2021 compared with 8% in 2014 (95% CI: 8%-9%). From 2014 to 2021, staff who self-identify as a woman increased from 72% (95% CI: 71%-74%) to 76% (95% CI: 75%-77%). Overall, 22% (95% CI: 21%-23%) of the SHA workforce rated their mental health as poor/fair. Conclusion: The 2021 PH WINS results represent unique and current perspectives on the SHA workforce and can inform future public health infrastructure investments, research, and field practice to ensure a strong public health system.