Recent research conducted within the veterinary profession has reported higher rates of depression and stress than the general US population. While this decline in mental well-being has been documented in Doctor of Veterinary Medicine (DVM) students and veterinary professionals, there is a lack of research on the mental well-being of the pre-veterinary population. This gap led us to conduct a survey in the fall of 2021 utilizing the DASS-21 and ATSPPH-sf inventories to assess the levels of depression, anxiety, stress, and help-seeking stigma in pre-veterinary students to better understand when the decline in veterinary mental well-being begins. A pre-test survey was completed by 233 pre-veterinary students in September, and an identical post-test survey was completed by 184 pre-veterinary students in November. From the pre- and post-test data, depression, anxiety, and stress scores increased as students advanced in academic status during their undergraduate degree. Juniors reported the highest averages of depression, anxiety, and stress compared with their peers. In the post-test, sophomores and juniors exhibited higher rates of depression than freshmen, and juniors and seniors exhibited higher rates of stress than freshmen. Current VMCAS applicants exhibited higher levels of stress than non-VMCAS applicants in the pre-test and lower levels of stress in the post-test. In both the pre-test and post-test data, respondents averaged a neutral attitude toward help-seeking. Based on these results, a decline in pre-veterinary mental well-being occurs as students' progress in their undergraduate career and should be further studied to assess its impact on Doctor of Veterinary Medicine and veterinary professional well-being.