The present report describes two outbreaks of serious enteritis in commercial laying hens where Clostridium perfringens and Clostridium colinum were simultaneously detected. At the age of 44 and 31 weeks, two laying hen flocks showed an increase of the mortality rate and a worsening of productive performance. Post-mortem examination revealed intestinal necrotic-haemorrhagic ulcerations and hepatic focal necrosis. The bacteriological examination yielded the isolation of C. colinum and C. perfringens toxin type A, NetB positive. In one outbreak, C. colinum was detected also by polymerase chain reaction in all the intestines of affected birds. In laying hens, C. colinum has never been isolated but only suspected as the causative agent of a slight enteric disease called duodenal focal necrosis. The present case report was characterized by severe enteritis presumably due to the synergistic effect of C. colinum and C. perfringens.