Brucellosis represents a serious problem in cattle relaying huge economic losses. Brucella meletensis on of brucella strains that can infect human which its usual host is sheep, might isolated from cattle. The aim of present study was detection of brucellosis among cattle population in Menoufia governorate, Egypt and analyze risk factors. Serologically brucella were detected with RBPT. Evaluation of the specificity and sensitivity of RBPT, IC assay and cELISA against CFT test were done. Molecular detection of brucella using PCR and bacteriological isolation followed by phenotypic and molecular typing of the isolated bacteria. The overall prevalence using RBT was 1.44%. Holstein Friesian cattle have highest prevalence (1.57%) while native cattle were (1.28%). Cattle over 3 years old have the highest seroprevalence (2.77%). The positive result for RBPT, cELISA, IC Assay and CFT were 91%, 60%, 91% and 88% respectively after examination 100 serum sample. Brucella melitensis biovar 3 was isolated from the tissue specimens (uterus and/or lymph nodes). PCR targeting (Immuno-dominant antigen, gene bp26) generated product of 450 bp from (16/20) tissues specimens. These findings assisting in future planning pragmatic control strategies against bovine brucellosis in Egypt and for herd health fertility.