One hundred and eighty raw milk samples were collected randomly from dairy farms, farmers` houses and dairy shops in Assiut city. These samples represented by 60 milk samples from cows and buffaloes of dairy farms (30 for each) , 90 samples from farmers` houses including cows, buffaloes and sheep milk (30 for each) and 30 raw dairy shops` milk samples. The samples were examined for the incidence of T.pyogenes and prevalence of Subclinical Mastitis (SCM) in raw milk. The prevalence of SCM in milk samples of dairy farms were 90% for both cows and buffaloes` milk based on California Mastitis Test (CMT) and 50% and 46.7% in cows and buffaloes` milk based on White Side Test (WST), respectivey. In the contrary, prevalence of SCM in farmers` houses milk samples based on CMT and WST were 53.3%, 73.3% &33.3% and 33.33%, 46.7% & 50% in cow, buffalo and sheep, respectively. The incidences of T.pyogenes in dairy farms milk samples were 60% in both cows and buffaloes` milk, while 63.33%, 60% and 36.67% in cows, buffaloes and sheeps` milk of farmers` houses, respectively. Additionally, the incidence of T.pyogenes in dairy shops` raw milk samples was 43.3%. PCR for 18 strains of T.pyogenes revealed that 15 strains gave positive results for pyolysin virulence gene. The public health hazards of the organism and the suggestive measures were also discussed.