Five bacterial strains (SYSU YG23(T), SYSU 10HL1970(T), 10HP82-10, 10HL1938, 10HP457) isolated from water reservoirs of cooling systems were characterized using a polyphasic taxonomic approach. The isolates were Gram-stain-negative, strictly aerobic and non-motile. Growth was enhanced in the presence of L-cysteine. The major fatty acids (> 5 %) for the five strains were C-10:0, C-16:0, C-16:0 3-OH, C-18:0 3-OH and C-18:1 omega 9c. Ubiquinone-8 was detected as the respiratory quinone while the polar lipid profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, three unidentified phospholipids, two unidentified aminophospholipids and three unidentified glycolipids. The strains shared 16S rRNA gene sequence similarities of 99.0-99.2 % with Francisella guangzhouensis 08HL01032(T) but less than 95.2 % with other members of the family Francisellaceae. The phylogenetic dendrogram based on 16S rRNA gene sequences showed that these strains form a separate cluster along with Francisella guangzhouensis. This cluster was also confirmed from multilocus-sequence typing based on sequences of the mdhA, rpoB and sdhA genes. Matrixassisted laser desorption ionization time-of-flight MS analyses of the strains along with closely and distantly related Francisella strains also showed a distinct cluster for these strains. Based on the findings from the polyphasic taxonomy studies, the strains were considered to represent two novel species of a new genus for which the names Allofrancisella inopinata gen. nov., sp. nov. (type strain SYSU YG23(T) = KCTC 42968(T) = DSM 101834(T)) and Allofrancisella frigidaquae sp. nov. (type strain SYSU 10HL1970(T) = KCTC 42969(T) = DSM 101835(T)) are proposed. In addition, Francisella guangzhouensis Qu et al. 2013 is proposed to be transferred to this new genus as Allofrancisella guangzhouensis comb. nov.