A total of 1317 samples were collected; 1164 serum samples, 122 milk samples, 24 lymph nodes and 7 aborted foeti from buffaloes in 10 Governorates from farms and villages in Upper Egypt. The serological tests used for the diagnosis of brucellosis on blood sera were the Rose Bengal plate (RBT) , Buffered acidified plate antigentest (BABAT), EDTA modifiedstandard tube agglutination test (MSAT), Revanol test (RT). On the other hand, the milk ring test (MRT) was performed on buffalo-cow's milk. Suspected colonies were stained with Gram, s stain and Modified ZeilNeelson stain. The isolated Brucella organisms on antibiotic free Brucella agar medium were subjected to the following tests for biochemical identification tests as CO2requirement, H2S production, Urease activity, growth in the presence of dyes,The indirect solid phase ELISA technique was carried out according to serum and milk samples.Agar gel immune diffusion test (AGID) and PCR applied on isolated Brucella strains. The results of the serological tests wereRose Bengal test 34.7%, BAPA (37%), Revanol test (28.2%),modified SAT (23.7%), indirect ELISAwere (32.3%) and AGPT (33.8%)in this study.Brucellaorganisms from lymph nodes of slaughtered buffaloes by culturing method showed that 3 (13.64%) isolates(2) of B. melitensisbiovar 3 and (1)B. abortusbiovar1. The isolated strain from aborted foeti was one isolate (14.29%) typed as B.melitensisbiovar 3. isolated only from Beni-Suef.By milk ring test (MRT) milk samples were 10 (8.20%) of B. melitensis biotype 3. A multiplex was
format that will allow the rapid identification of Brucella spp., B. abortus,
and B.melitensis in a single test within 2 to 3 h. B. melitensis was identified
at 731bp and B. abortus identified at 498bp. Finally, we made measures of
the control program for eradication of brucellosis in buffaloes by a
reasonable system of compensation, Veterinarians for field work and state
laboratories capable of serological techniques.Also, information technology
solutions and further logistic means as animal identification techniques are in
many governorates in Egypt.