The objective of this study was to identify the presence of Neospora caninum by polymerase chain reaction (PCR), in brain, myocardium and liver (150 organs) obtained from 50 free-range chickens (Gallus domesticus) that coexisted with bovines and dogs in Central Mexico. N. caninum DNA was detected in two brains out of 150 organs (1.3%) from two out of 50 birds (4%). Results from the present study suggest that free-range chickens (Gallus domesticus) of Central Mexico are infected of N. caninum. Further, although these parasites were detected in only a small percentage of birds (4%), they may represent a potential risk factor for dogs that feed on them and eventually excrete oocysts, thereby contaminating the water and food of other intermediate hosts. This finding is of particular importance for cases of horizontal transmission in cattle.