This investigation aimed to knowledge of the presence of Brucella spp .in the samples from Babylon province and the relationship with environmental factors. The seasonal collection of animal soil samples for various villages in the middle, south, and north of Babylon province, Hilla City, Iraq, with physicochemical parameters (air and soil temperature, pH, EC, TDS, salinity, TOC), as well as heavy metals (Fe, Cu, Cd, and Pb). The results showed the presence of Brucella species in some of soil samples and its percentage varies according to the four seasons, where the highest percentage of Brucella spp. was in summer season and the lowest in winter season and these percentages were related to seasonal environmental indicators. After sending the positive bacterial samples isolated from the soil for sequencing analysis, it was found that the bacterial samples isolated from the soil have new strains recorded in Genbank as following: five strains for Brucella intermedia and one strain for Brucella rhizosphaerae, Also, phylogenetic analysis was for the drawing of phylogenetic tree and the determination of phylogenetic relationship for Knowledge the degree of convergence between species and their source. Through our research, we concluded that proportion of Brucella spp. presence in summer was high and also environmental factors which included (Air and Soil temperature, pH, EC, TDS and Salinity) were high, except TOC was lower in summer season whereas heavy metals for Cu, Fe, Cd were lower in summer except Pb was high. This indicated that differences of seasons affect to Brucella ratio in soil. The findings of sequencing for the bacterial samples from soil demonstrated that B. intermedia was found in both the spring and fall seasons while B.rhizosphaerae found in the spring season , These types of Brucella had genetic variations. Also concluded that during phylogenetic analysis of the 16S rRNA sequences indicated that B.intermedia was nearest to Pakistan and other strain of B.intermedia was nearest to Russia while B.rhizosphaerae was nearest to Ukraine and Pakistan.