The objective of the current study was to determine the effect of pre-slaughter stress, season and breed on the activity of plasma creatine kinase (CK) and the quality of mutton. One hundred and seventy-three (173) castrated sheep from Dormer (DM), South African Mutton Merino (SAMM), Dorper (DP) and Blackhead Persian (BP) sheep breeds were used in the study. The animals were grouped according to age-groups as follows: Group 1 (6 to 8 months), Group 2 (9 to 12 months) and Group 3 (13 to 16 months). Blood samples were collected during exsanguinations using disposable vacutainer tubes for CK analysis. Representative samples of the Muscularis longissimuss thoracis et. lumborum (LTL) were collected from 84 castrated sheep, of different breeds (28 per breed) 24 h after slaughter. The following physico-chemical characteristics of mutton were determined; meat pH ($pH_{24}$), color ($L^*$, $a^*$ and $b^*$), thawing and cooking losses and Warner Braztler Shear Force (WBSF). The activity of plasma CK was significantly higher (p<0.001) in summer ($1,026.3{pm}105.06$) and lower in winter ($723.3{pm}77.75$). There were higher values for $L^*$ ($33.7{pm}0.94$), $b^*$ ($11.5{pm}0.48$) and WBSF ($29.5{pm}1.46$) in summer season than in winter season; $L^*$ ($29.4{pm}0.64$), $b^*$ ($10.2{pm}0.33$) and WBSF ($21.2{pm}0.99$). The activity of plasma CK was influenced by the type of breed with Dormer having the highest (p>0.001) levels ($1,358.6{pm}191.08$) of CK. South African Mutton Merino had higher values for $pH_{24}$ ($5.9{pm}0.06$), $L^*$ ($34.2{pm}0.97$), $b^*$ ($12.2{pm}0.50$) and WBSF ($26.8{pm}1.51$) and Blackhead Persian had higher values ($35.5{pm}2.17$) for cooking loss (CL%) than the other breeds. Computed Principal Component Analyses (PCA) on the activity of plasma CK and physico-chemical characteristics of mutton revealed no correlations between these variables. However, positive correlations were observed between $pH_{24}$, $L^*$, $a^*$, $b^*$, CL% and WBSF. Relationships between pre-slaughter stress, CK activity and physico-chemical characteristics of mutton were also observed. It was therefore concluded that although mutton quality and creatine kinase were not related, pre-slaughter stress, season and breed affected the activity of creatine kinase and mutton quality.