This study was conducted to evaluate the effect of pre-slaughter handling on lamb welfare and meat quality of Merino lambs exposed to different pre-slaughter handling conditions. The objective of this study was to investigate the effect of transportation distance and lairage duration on creatine kinase (CK), lactate dehydrogenase (LDH), carcass and meat quality from Muscularis longimissius thoracis et lumborum (LTL) characteristics Merino lambs slaughtered at a commercial abattoir. Data was collected from eight-month old female Merino lambs (n=100). Exsanguination blood for the analysis of the activities of CK and LDH were collected using vacutainer tubes treated with EDTA. Carcass measurements, pHu, temperature and meat colour coordinate readings were carried out 24 hours after slaughter. Furthermore, thawing loss (TL), cooking loss (CL) and warner braztler shearforce (WBSF) was measured from the LTL after 7 days post slaughter storage (-20 ºC). Lambs that travelled for a longer distance had higher levels of LDH compared to those that travelled a shorter distance. Lambs transported for a longer distance also had a higher TL and tougher meat. Lambs placed at the lairages for longer duration at the lairages had a higher cooking loss compared to the shorter duration. The results indicate that longer transportation and lairage durations made the lambs more susceptible to pre-slaughter stress and negatively affected the meat quality.