Streptococcus gallolyticus (S. gallolyticus) infection is considered an important septicemic disease in pigeons especially in squabs. In this study, 145 different samples from old pigeons and squabs from internal organs (liver, lung, heart blood and kidney. Samples were collected from pigeon's sailing shops at Cairo and Giza governorates. Samples were cultivated for
bacteriological isolation targeting streptococci especially the so called S. gallolyticus. Morphological, biochemical and molecular identification of bacterial isolates revealed presence of S. gallolyticus with total incidence of 11.72%. The individual incidences were 12% (El Marg), 9% (Ein shams), 8% (Nasr city), 17% (El- Moneeb), (11% (El- Hawemdia) and 12%
(El- Badrashin). Isolates were subjected to antibiotic sensitivity test and multiplex PCR targeting two virulence genes namely ScpB and Lmb as well as 5 antibiotic resistance genes (Pbp1A, ermB, tetO, aac (6') and aph (2'').