Dairy cows are usually culled and transported from the farm when they no longer meet the farm’s standards for production or are not needed for milk production. Some cows are transported while in poor condition and may deteriorate further during transport. In February 2020, Canadian federal animal transport regulations were revised with the aim to minimise risks to livestock during transport; changes that may impact cull dairy cows included defining compromised cattle and limiting their maximum transport time. This study conducted semi-structured interviews with dairy farmers (n = 6) and cattle haulers (n = 4) in British Columbia, Canada, to gain an in-depth understanding of the effect of the regulations on their practices when shipping and transporting dairy cows to slaughter. Interviews were transcribed in Otter.ai and thematically coded in NVivo 12. While farmer and hauler participants recognised the importance of animal welfare during transport and described practices such as shipping mobile animals to reduce the risk that cows would become non-ambulatory during transport, they also described little change in shipping and transport practices due to the new regulations. Among interviewed participants, barriers to compliance with the regulations appear to be low knowledge of, and mixed or negative attitudes towards the regulations. Participants also described how they felt a lack of communication along the transport chain and limited transport and slaughter infrastructure made compliance difficult. Possible suggestions to remedy these barriers include providing educational resources about the regulations and encouraging communication about cow fitness for transport between responsible parties in the transport chain.