Castration has been demonstrated to cause pain in sheep. However, it is routinely performed for management purposes. Electroencephalography (EEG) has been used successfully to measure pain in lambs in response to castration and other husbandry procedures in livestock. The aim of this study was to evaluate the use of EEG as a measure of pain and analgesia in conscious lambs undergoing castration on farm over a 24 h period. EEG responses were compared to behavioural observations and changes in ocular temperature via infrared thermography. Twenty-four merino ram lambs (18.63 ± 2.06 kg) were used in this study. Lambs were randomly allocated to one of the following treatment groups: (1) castration with pre-surgical administration of meloxicam (0.5 mg/kg Metacam 20 mg/mL injected subcutaneously into the skin of the neck 15 min prior to recording) and lignocaine (applied via intra-testicular injection five minutes prior to castration, 2 mL lignocaine hydrochloride 20 mg/mL, Troy/Ilium) (CML, n = 8); (2) castration only (C, n = 8); (3) sham castration, handling only (SC, n = 8). EEG was recorded for 5 mins pre-procedure (prior to any intervention), and for 5 mins post-procedure at 0, 1, 4 and 24 h. Behavioural reactions to the procedure were scored, and behaviours were scan sampled at 5 min intervals at the above time points, by blinded observers. Eye temperature was measured for five-minute intervals at each time point. EEG decreased from baseline to 0 h for CML and C groups (p < 0.001), C group values returned similar to baseline at 24 h. Eye temperature increased post-castration at 0 h for C group, no initial change was seen for CML or SC groups. CML and C groups were more likely to have higher reaction scores and showed more abnormal behaviours (p = 0.017). CML and C groups had similar results, indicating minimal effect of analgesic intervention. Lambs in SC group showed significant EEG changes, suggesting that stress from handling also impacted EEG results.