Extensive use of per- and polyfluoroalkyl substances (PFAS) has resulted in their ubiquitous presence in human blood. PFAS exposures have been associated with multiple adverse human health effects. Biomonitoring studies have focused on long-chain PFASs, but these are being replaced by short-chain PFASs or with alternate PFAS chemistries (or replacement chemistries such as GenX), resulting in changes in human exposures with time. Here, we take advantage of serum samples collected as part of a clinical trial testing the efficacy of a dietary fiber intervention to reduce serum cholesterol to investigate exposure to PFASs in Canadian participants. Serum samples were collected from 72 participants (adult males with elevated cholesterol) in 2019-2020 at baseline and after 4 weeks of the intervention and were analyzed for 17 PFASs. The highest geometric mean serum concentrations of PFAS measured at baseline corresponded to PFOSA (7.1 ng/ml), PFOS (4.2, ng/ml), PFOA (1.8 ng/ml) and PFHxS (1.3 ng/ml). Four long-chain PFASs (PFOA, PFOS, PFOSA and PFHxS) and two short-chain PFASs (PFBA, PFHxA) were detected in 100% of participants. GenX was detected in 71% of participants. Analyses of associations between serum-PFAS concentrations and biomarkers of adverse health outcomes showed the PFBA, PFHxA, PFDA and PFOSA were associated with higher serum gamma-glutamyl transferase concentrations but not with measures of serum-total or low-density lipoprotein cholesterol. Comparison of PFAS concentrations at baseline and after a 4-week follow-up showed that the total PFAS detected decreased in both the control and cholesterol intervention groups. However, the suite of long-chain PFASs of concern identified by the United States National Academies of Sciences, Engineering, and Medicine, significantly decreased only in the cholesterol intervention group. This observation suggests that a sustained dietary fiber intervention may reduce long-chain PFAS body burden, but future intervention studies need to control for PFAS exposure sources and extend the dietary supplement intake beyond 4 weeks. Overall, the results show that exposures to short-chain and replacement chemistry PFASs are common in this Canadian population.