Clostridium perfringens is a major necrotic enteritis causing bacterial pathogen in poultry, and a source of food poisoning and gas gangrene in humans. C. perfringens can also cause mild to severe enteritis in pigs. In the EU, the occurrence of C. perfringens associated necrotic enteritis in poultry has increased as antibiotic use has decreased. As the US moves away from the use of antibiotics in animal feed, we can expect an increase in necrotic enteritis with subsequent losses from morbidity and, in subclinical cases, losses from decreased chicken weights. Alternatives to antibiotics in animal feed will be needed in the near future. The genomes of 43 C. perfringens isolates from chicken were sequenced, and examined for peptidoglycan hydrolase enzymes by homology to known enzymes. There were hundreds of putative peptidoglycan hydrolases identified that clustered into 15 groups from 3 families common to endolysins as identified by PFAM and phylogenetic analysis. Members of each family were aligned and split into groups by phylogenetic tree analysis (7 amidase_3 groups, 4 amidase_2 groups, and 4 GH25 groups). Members from 10 different groups were cloned and expressed in E.coli. Four lysins were shown to have high lytic activity against all 43 of the initial isolates in plate lysis assays but not other Gram positive or Gram negative species tested. Activity was also demonstrated in zymogram and turbidity reduction assays. One lysin was shown to have an internal translational start site producing two products when expressed in E. coli. The domain architecture and relative activity of the four lysins will be discussed.