## 3 Results and Discussion  
Pre-slaughter stress during transport, lairage, and slaughtering may play a decisive role in determining beef quality. After stressful transport incidents, resting time is essential for glycogen depletion, tissue rehydration, and electrolyte restoration [37; 38]. In Europe and North America, cattle slaughtering activities generally occur on the same day that the animals arrive to the abattoir. This guarantees agile carcass management and shorter processing times, which might eventually increase meat quality [24]. Implementing this system can potentially prevent negative behavioral reactions resulting from prolonged lairage time, especially when animals suffer from dehydration and feed deprivation, which can contribute to further glycogen decline [38; 39]. It was established that even if animals had ad libitum access to food and water, stress reactions associated with exposure to new environments and new personnel might decrease their necessity for drinking and eating [24]. The mixing of unfamiliar animals before slaughter can also compromise meat quality, especially when cattle are repeatedly loaded and unloaded [28]. It has been reported that the meat from cattle commercialized through saleyards and auctions often exhibits higher pH and lower consumer acceptability than the meat from animals directly sold to the abattoir. It is possible that these animals are more often exposed to multiple stress situations caused by strange environments, animals, and operators [40; 41]. It has also been argued that cattle exhibit more aggressive behavior when mixed during pre-mortem operations, which increases the probability of finding carcass bruises, thus affecting the carcass output and compromising meat quality [41; 42]. Keeping animals between their familiar partners during finishing, loading, transporting, and slaughtering has been demonstrated as providing a calming effect for the animals, which can result in improved meat quality [38; 43]. Another reason for increased pre-slaughter stress relates to the exposure of animals to extreme cold or hot environments. Cold weather combined with precipitation, along with large temperature fluctuations during slaughtering days, might increase involuntary movements experienced by animals (shivering) [44]. It has been hypothesized that these involuntary movements could lead to more intense mitochondrial biogenesis and cause higher mitochondrial oxygen consumption and lower color values in fresh meat [32]. Negative effects on the physical and physiological status of the animals have been linked to slaughtering temperatures higher than 21 \({}^{\circ}\)C or lower than 5 \({}^{\circ}\)C [45; 46]. A previous study showed that up to 30% of the meat of young bulls slaughtering the summer season had pH \(\geq\) 6.2, which denoted DFD-related issues [47]. In our experiment, the average daily temperatures throughout the slaughtering days were between 10 and 18.8 \({}^{\circ}\)C, with temperature fluctuations no higher than 8 \({}^{\circ}\)C, which probably contributed to the observation of meat samples with pH\({}_{48}\)\(<\) 5.8 in all treatments. Combined Effects of Transport Distance (TD), Animal Weight (AW) and Muscle Position (MP) on Meat Quality Traits  
The results of the combination of factors yielded a statistically significant difference (\(p<0.01\)) between different TD, AW, and MP levels using the Pillay's Trace test. Based on these findings, there was enough evidence to show that TD, AW, and MP influenced meat quality parameters, as shown in Table 3.  
The interaction between factors resulted in only significant differences for AW \(\times\) TD. This combination of factors indicated that heavy animals (AW \(>\) 445 kg) transported over medium distances (34.5 \(<\) TD \(<\) 59.1 km) produced meat with the highest average pH\({}_{48}\) values, and this combination also yielded steaks with the lowest hardness scores from all treatments, according to the results from the sensory analysis.