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Twenty-one small Gram-negative motile coccobacilli were isolated from 15 systemically diseased African bullfrogs ( Pyxicephalus edulis ), and were initially identified as  Ochrobactrum anthropi  by standard microbiological identification systems. Phylogenetic reconstructions using combined molecular analyses and comparative whole genome analysis of the most diverse of the bullfrog strains verified affiliation with the genus  Brucella  and placed the isolates in a cluster containing  B. inopinata  and the other non-classical  Brucella  species but also revealed significant genetic differences within the group. Four representative but molecularly and phenotypically diverse strains were used for  in vitro  and  in vivo  infection experiments. All readily multiplied in macrophage-like murine J774-cells, and their overall intramacrophagic growth rate was comparable to that of  B. inopinata  BO1 and slightly higher than that of  B. microti  CCM 4915. In the BALB/c murine model of infection these strains replicated in both spleen and liver, but were less efficient than  B. suis  1330. Some strains survived in the mammalian host for up to 12 weeks. The heterogeneity of these novel strains hampers a single species description but their phenotypic and genetic features suggest that they represent an evolutionary link between a soil-associated ancestor and the mammalian host-adapted pathogenic  Brucella  species.

Brucella spp. of amphibians comprise genomically diverse motile strains competent for replication in macrophages and survival in mammalian hosts