Effect of pen size on behavioral, endocrine and immune responses of water buffalo (Bubalus bubalis)

Female water buffalo ( Bubalus bubalis) calves (n = 28) aged 7 to 10 d were divided into four groups of seven animals each to examine the effects of space allowance (Group A: 2.6 indoor m2 + 2.0 outdoor m2/calf; Group B: 2.6 indoor m2/calf; Group C: 1.5 indoor m2/calf; Group D: 1.0 indoor m2/calf) on behavioral, endocrine, and immune variables for a period of 60 d. Animals were offered 7 L/d of a commercial acidified milk substitute. The calves averaged 45.9 kg initially and 92.4 kg finally. The behavior observations were conducted 7 d after grouping and fortnightly thereafter. At wk 4 and 8, the phytohemagglutinin (PHA) skin test was performed to induce aspecific delayed hypersensitivity. At wk 1 and 3, calves were injected i.m. with keyhole limpet hemocyanin. Antibody titers were determined at weekly intervals for 7 wk. Calves in pens with greater space allowance (Groups A and B) were less active than Groups C and D ( P < .001). The latter groups were also observed feeding more often at wk 7 ( P < .01). Calves provided with an outdoor paddock spent less time standing than Groups C and D ( P < .01), and lay with a greater number of outstretched legs ( P < .001). Groups C and D showed a lower reaction to PHA in both skin tests than did Groups A and B ( P < .001 and P < .05, respectively). Group A showed an antibody response consistently higher than groups B, C, and D ( P < .01, P < .05, and P < .05, respectively). At the end of the experimental period, the calves were subjected to an isolation test lasting 10 min. Group D showed a longer duration of movement with respect to Groups A and B ( P < .01); animals from Group C walked more than did Group A ( P < .05). Cortisol concentration evaluated 0, 10, 45, 90, 150, and 225 min after separation from the group was higher in Groups C and D than in Groups A and B ( P < .01). For all animals, the highest cortisol level was observed immediately after the isolation test ( P < .001). Space restriction resulted in evidence of stress in the animals as shown by alterations in a number of physiological responses. However, the use of small groups of only seven animals per pen may have affected their reactions to space restriction. It is possible that using larger groups could change these conclusions.