In the present study, the rôle of gammaglutamyltransferase (GGT) during lactation has been investigated in the water buffalo. GGT activity has been evaluated in the mammary tissue at 4 and 6 months after calving and during the non-lactating period. The highest GGT activity levels were found at day 120 (32·57±7·41 U per g) of lactation and were statistically higher than those at 180 (10·76±3·6 U per g) or during the non-lactating period (9·86±7·94 U per g). Histochemistry confirmed these findings and revealed that GGT reactivity was distributed throughout the cytoplasm of alveolar epithelial cells. Such results showed that the GGT production is high during lactation thus supporting the hypothesis that this enzyme plays a rôle in determining milk production in water buffalo by supporting milk protein synthesis.