*2.3. Behavioural Response*

The behaviour recordings during loading at the farm and unloading at the abattoir included slipping, falling, reluctance to move, turning back, overlapping, and vocalization as previously described [22,23]. Lying, sitting, and standing behaviours after 25 min of resting time was directly observed for a period of 2 min.

#### *2.4. Blood Sampling and Analysis*

The animals were blood sampled five days before delivery as reference for basal blood parameter level (T0) and at exsanguination (T1) for the analysis of creatine kinase (CK), cortisol, glucose, lactate, albumin, albumin/globulin ratio, total protein, urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphate (ALP), sodium (Na−), and potassium (K+). All biochemical parameters except for glucose, were measured in serum obtained from blood collected in serum separator tubes with gel separator and clot activator (Vacutest Kima, Padua, Italy), let to clot and centrifuged at 1300× *g* at 20 ◦C (Eppendorf 5702R, Eppendorf, Milan, Italy) for 10 min. Plasma for the measurement of glucose was harvested from anticoagulated blood collected in Na2EDTA test tubes containing the glycolysis inhibitor potassium fluoride (Vacutest Kima, Padua, Italy), centrifuged at 1300× *g* at 20 ◦C for 10 min. Parameters were measured by colorimetric assays on automated analyser (Olympus AU 400, Beckman Coulter, Milan, Italy) with the exception of cortisol. Serum cortisol was determined using the Kit Immulite One Cortisol (medical system code LKC01, Siemens Health Care Diagnostic Limited, Glyn Rhonwy, Gwynedd, UK).

## *2.5. Skin Bruises Measurement*

Carcasses were horizontally exsanguinated for 5 min and then hung for 10 min before being submerged in a scalding tank for dehairing at 62 ◦C for 8 min. After dehairing, skin damages were subjectively assessed by a trained observer, using a four-point scale (1 = none to 4 = severe) based on the scale developed by the Danish Meat Research Institute (DMRI) [24]. The DMRI scale was used to score all skin lesions on the front (head included), middle and hind quarters of each carcass. Moreover, a skin damage score for the whole carcass was calculated using the highest score assigned to each quarter [24]. At about 40 min post mortem, carcasses were split, weighed, and transferred to the chilling room.

## *2.6. Meat Quality Measurements*

Measurements of pH on the *longissimus thoracis* muscle (LT) at the level of six/seven thoracic vertebra were recorded at 1, 3, and 6 h post mortem on the left side, with a pH-meter (mod. HI8424, Hanna, Padua, Italy) equipped with Crison electrode (Crison Instruments, SA, Barcelona, Spain) and an automatic temperature compensation probe. At 6 h post mortem the left side was sectioned in the primal cuts and a sample of LT muscle between 6 and 10 thoracic vertebra (10 cm thickness) was collected. After 30 min of blooming, L\*, a\*, and b\* coordinates were determined using Minolta chroma-meter (CR-300, Minolta Camera Co., Ltd., Osaka, Japan) with D65 light source and 0 ◦C viewing geometry. Samples were transferred (by air) to a laboratory of the Department of Agricultural and Food Sciences (Bologna, Italy). Measurements of pH were repeated at 24 and 72 h post mortem and colour measurements were repeated at 24, 72, and 144 h after slaughter. At the same time interval, cooking loss was determined on a slice of the LT muscle according to Honikel [25] and Warner-Bratzler shear force (WBSF) was measured after cooking using Instron apparatus (mod. 1140, Instron, Norwood, MA, USA). Drip loss was determined at 24 h post mortem [25].
