The objective of this experiment was to determine the effects of preslaughter isolation and feed withdrawal duration on physiological responses and shrinkage in goats. A total of 84 Spanish does (36 mo of age, average weight 35 kg) were individually weighed and scored for excitability before two replicate (day) trials. The does were feed-deprived (FD) or fed (F) in holding pens (treatment, TRT) for either 0, 7, 14, or 21 h (TIME). At the end of the holding periods, FD and F does were blood-sampled (n = 6 does/treatment/time/replicate) and weighed again to assess physiological responses and shrinkage, respectively. Individual does from each pen were blood-sampled again after imposing one of three handling post-treatments: a 15-min isolation with no visual contact with other does (I); a 15-min isolation with visual contact (IV); or no isolation (C, control). Plasma cortisol concentrations were higher at 0 h than at other holding time periods (P < 0.01). Plasma triiodothyronine, thyroxine, and leptin concentrations, and differential leukocyte counts were not influenced by any of the factors. The rate of decline in glucose concentrations over TIME was greater in FD than in F group (TRT x TIME, P < 0.05). The overall plasma creatine kinase activity peaked at 7 h before reaching a lower level at 14- and 21-h holding (P < 0.05). Plasma urea nitrogen concentrations were higher at 0- and 21-h than at 7- and 14-h holding (P < 0.01). Plasma nonesterified FA concentrations in the FD group remained at an elevated level during holding, but in the F group the levels decreased at 7 h and remained at that level (TRT x TIME, P < 0.01). Excitability scores did not have any effect on the variables measured. Shrinkage increased with longer holding time, but more prominently in the FD group (TRT x TIME, P < 0.01). Plasma cortisol concentrations were greater in I and IV groups than in the C group (P < 0.01). The novelty of environment during preslaughter holding, and social isolation may be more potent stressors than feed deprivation in goats, although shrinkage may increase with increasing feed-withdrawal times.