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Simple Summary: Journey duration, position of the animal in the vehicle and time spent in the lairage facilities before slaughtering can negatively affect pig welfare. The effects of journey duration and deck level on hormonal parameters were examined in this pilot study during three commercial truckloads. Plasma cortisol, epinephrine and norepinephrine increased with longer transportation time. The levels of the tested hormones were higher in pigs transported for 11 h than for 6.5 h or only 3 h. The loading position (deck level) of the animals on the truck strongly affected only norepinephrine. While the effects of journey duration on plasma cortisol levels were expected and in line with the literature, our findings on plasma catecholamines are novel and may be useful to monitor transportation stress responses in pigs. These preliminary findings need to be confirmed by future studies. The aim of this pilot study was to evaluate the relationship between journey duration, deck level and activation patterns of the hypothalamic-pituitary-adrenocortical axis (HPA) and sympathetic adrenal medullary system (SAM) in pigs. A total of 90 pigs were examined. The animals came from three different Italian farms associated with the same slaughterhouse located in Bari (Apulia region-Italy). A group of thirty animals was transported from Pordenone (11 h journey); a second group was transported from Terni (6.5 h journey); a third group was transported from Benevento (3 h journey). The animals were transported in the same vehicle, which complied with the structural characteristics indicated in the Council Regulation (EC) No. 1/2005. The truck was composed of a lorry and a trailer, each one divided into three decks. Only the animals transported in the trailer were tested for the study. Before transportation, blood samples were collected on each farm, at 6:00 a.m., from 30 pigs randomly selected out of 135 pigs ready to be transported. Blood samples were also collected during slaughter to evaluate plasma cortisol, epinephrine and norepinephrine, around 6:00 a.m. A journey duration of 11 h was associated with significantly higher plasma concentrations of stress hormones compared with shorter journeys. This increase was proportional to the journey duration, with the pigs travelling for 6.5 h displaying intermediate concentrations between those noticed after 3 h and 11 h journeys. The interaction between deck and journey distance was not significant on epinephrine, norepinephrine or cortisol levels collected at arrival. There was a significant effect of deck level on norepinephrine levels (p < 0.0001), a tendency to influence epinephrine levels (p = 0.073) but no effect on cortisol levels (p = 0.945). Overall, we observed that an 11 h-long journey seemed to impact negatively on pigs' HPA-SAM activity, likely requiring the animals to spend more time in the lairage facilities to recover. [ABSTRACT FROM AUTHOR]