A comprehensive stress assessment is vital in understanding the impact of the pre-slaughter procedure on animal welfare. The transportation and handling process was commonly reported to cause stress in animals. This research utilises electroencephalography (EEG) as an alternative stress indicator to non-painful acute stress measurement. EEG has been proved to be instantaneous and sensitive with specific results. Therefore, this study was aimed to determine the stress level of goats subjected to two different transportation duration and the effect of lairage based on their EEG activities and blood parameters changes. Eighteen adult male goats were divided into two transportation stress groups based on the transport duration: the two-hour (TS2) and six-hour (TS6) groups. Then, each group was then again divided into three smaller groups according to the lairage duration, which was three-hour (L3), six-hour (L6), and overnight (L12) groups. Blood was sampled before transport, after transport, and during slaughter while EEG was recorded before transport, after transport, after lairage, and during slaughter. Results revealed that there was a significant decrease in beta wave activity compared to baseline in TS2 goats (P < 0.05) after transportation, whereas no significant difference was detected in the TS6 goats. At the same time, goats from the TS2 group showed increase in creatine kinase (CK) and lactate dehydrogenase (LDH) compared to that in TS6 goats. Together with the observed cortisol concentration, these findings showed that the TS6 goats were fully adapted to the transportation stress while the TS2 goats were still under stress. As for the lairage duration, it was observed that the TS2L3 goats showed lower EEG activities than the values obtained after two-hour transportation, while lower EEG activities were found from the TS6L6 goats after six-hour transportation. Therefore, it can be concluded that three-hour lairage was adequate to lower the impact of two hours transportation stress, whereas six-hour lairage was required to reduce the impact of six hours transportation stress. Finally, it was also found that the TS6L3, TS6L6, and TS6L12 groups took a long time to die after slaughter than the TS2L3, TS2L6, and TS2L12 goats based on the time their EEG activity reached isoelectric.

This preliminary trial investigated the effect of transportation and lairage periods on physiological parameters of goats subjected to slaughter. Nine male Boer cross goats aged 8-12 months were transported for 6 h and kept at lairage for 3, 6, or 16 h (n = 3). Blood samples were collected at pre-(pre-T) and post-transportation (post-T), and post-slaughter (post-S) for determination of hematological parameters, serum enzyme, protein, and cortisol concentrations. Electroencephalogram readings were taken at pre-T, post-T, pre-slaughter (pre-S), and post-S to determine the median frequency (F 50 ) and total power (Ptot) values. At post-T, there were manifestations of stress leukogram; increase in hematocrit, total protein, and muscle enzyme concentrations; and decrease in Ptot (p < 0.05). The high pre-T cortisol concentration suggests that the goats were already under stress before transportation. Stress leukogram became less evident after lairage, indicating that the goats had recovered from the stress of transportation. Although the Ptot increased at post-S especially following 3 h of lairage, F 50 values at post-S did not differ from pre-L, suggesting that the pre-slaughter stress may have affected the pain threshold. It is suggested that after 6 h of transportation, goats should ideally be placed in lairage for a minimum period of 3 h before slaughter.

Pain is a complex phenomenon, and to address pain management in animals, the presence of pain needs to be determined. Most studies assess the presence of pain based on the appraisal of either a single biomarker or a combination of a few parameters, which were often not examined simultaneously. In ruminants, measurements of cortisol and haptoglobin (Hp), and the assessment of behavioural changes have been the standard practice for this purpose. Nevertheless, there are still limitations in using these parameters, as they are also indicators of psychological stress, and can be misinterpreted as to indicate the presence of pain. Assessment of behavioural changes in ruminants can be difficult as prey animals conceal their experience of pain to avoid predators' attention.Therefore it is recommended that a reliable assessment tool be devised that includes a combination of various parameters to determine the presence of pain in ruminants in order to address pain management effectively.The primary aims of this study were to evaluate the conventional and contemporary biomarkers to determine the response of an animal to a noxious experience, and to refine a multi-parameter 'toolkit' that can be applied in assessing pain in ruminants. For these purposes, castration was used as a model of a common painful husbandry procedure that presumably evokes pain associated with the tissue injury.Merino cross lambs were assigned to two groups that were either surgically castrated or subjected to restraint stress (control). Castration was an open surgical technique without the provision of anaesthesia or analgesia. The conventional biomarkers estimated were plasma cortisol, Hp and betaendorphin (β-EP). The contemporary biomarkers measured were interleukin-6 (IL-6), substance P (SP), and prostaglandin E 2 (PGE 2 ). The peripheral leukocytes expression of IL-6 and proopiomelanocortin (POMC), the precursor of β-EP were also estimated. Behavioural changes were recorded to assess pain-related behaviour caused by the surgical procedure.Following treatment, the cortisol concentration in the castrated lambs was higher than control animals from ten minutes to two hours post-castration, indicative of the acute noxious experience associated with the surgical procedure. Surgical castration also caused a response in SP indicative of nociception and neurogenic inflammation, which was increased from 30 minutes and became significant eight hours following castration. The behavioural assessment showed that the castrated lambs demonstrated statue standing, a pain-like behaviour, from three to five hours post-castration, which were associated with the increase in SP, indicative of the pain sensitisation caused by the tissue injury. Increase in the systemic inflammatory mediators, the Hp and IL-6 were observed from ii day two, in which Hp remained significant until day four and resolved by day five. The IL-6 continued to increase over time, suggesting of an ongoing inflammatory response and inflammatory pain possibly until wound healing had occurre...