Hyperprolactinemia is associated with incubation behavior in avian species. Increased nesting activity is a major indication of incubation behavior. Vasoactive intestinal peptide (VIP) stimulates prolactin (PRL) secretion from the anterior pituitary. The goal of this study was to induce incubation behavior by stimulating PRL through chronically infusing VIP into the third ventricle of turkey brains. In experiment 1, porcine VIP (pVIP) was infused into the median eminence at a rate of 60 ng/min for 7 days by means of osmotic pumps implanted s.c.. Plasma PRL increased significantly in the pVIP-treated turkeys (p < 0.001). Although egg laying was not affected by the pVIP infusion, the mean oviduct weight decreased (p < 0.057). In experiment 2, saline or pVIP (30 or 60 ng/min) was infused into the third ventricle of laying turkeys for 12 days. Both pVIP treatments increased plasma PRL for 9 days (p < 0.05). The 30-ng pVIP/min infusion decreased nesting activity, plasma LH, ovary and oviduct weight, hypothalamic GnRH I, and anterior pituitary VIP receptors (p < 0.1). However, ovine PRL infusion (20.8 ng/min) into the same turkey flock increased nesting activity (p < 0.01). In conclusion, pVIP does not induce incubation behavior in laying turkeys.