Abstract The study objective was to determine if acute or chronic water restriction affected haptoglobin (Hp) and serum and mucosal antibody concentration in beef calves after parenteral or intranasal vaccination. A total of 60 crossbred beef steer (n = 28) and heifer (n = 32) calves were used. Animals were previously unvaccinated and confirmed seronegative for antibodies against infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea virus (BVDV), and parainfluenza-3 virus (PI3V) from sera collected at weaning (d -28; 183 days of age). On d -7, calves within sex were assigned randomly to 1 of 6 treatments arranged in a 3 × 2 factorial. Water restriction treatments (Factor A) were applied at the origin ranch via 3 different models: A1) Control, no water restriction except during transport to the feedlot (CON), A2) Acute, consisting of 48 h water restriction prior to transport to the feedlot (ACU), or A3) Chronic, consisting of alternating 24 h periods of water access and restriction, over a 7-d period prior to transport to the feedlot (CHR). Upon feedlot arrival (d 0), 2 respiratory vaccine treatments were applied (Factor B): B1) parenteral administration of a pentavalent modified-live virus (MLV) respiratory vaccine (2 mL s.c. in the neck; EX), or B2) intranasal administration of a trivalent MLV respiratory vaccine (1 mL/naris; IN). Sera was collected weekly from d 0 to 56 for antibody titer analyses, and d -7, -5, -3, -1, 0, 1, 3, 5, 7 and 14 to determine Hp concentration. Nasal swabs were collected on d 0, 3, 7, 14, 21, 28, 35, 42, 49 and 56 to determine presence of bovine respiratory syncytial virus (BRSV)-specific mucosal antibodies. Data were analyzed using Proc Mixed with repeated measures. Overall, haptoglobin was increased on d -7, decreased before feedlot transport on d -1, and increased after arrival (day effect; P < 0.001). An A × B × day interaction (P = 0.004) existed for Hp such that CON-EX and ACU-IN were greatest on d 5 (P ≤ 0.04). The BRSV (P ≤ 0.002) and IBRV (P ≤ 0.04) antibody titer was greater for IN vs. EX from d 14 to 56 and d 21 to 35, respectively. The EX treatment had increased BRSV-specific mucosal antibodies on d 3 and 7 (P ≤ 0.03); however, these were greater for IN from d 35 to 56 (P ≤ 0.05). In this study, water restriction did not clearly alter Hp or antibody responses to parenteral or intranasal vaccination but BRSV and IBRV antibody was increased for IN vs. EX.