Francisella asiatica sp. nov. isolated from farmed tilapia (Oreochromis sp.) and elevation of Francisella philomiragia subsp. noatunensis to species rank as Francisella noatunensis comb. nov., sp. nov.

Bacterial isolates from diseased farmed tilapia (Oreochromis sp.) from Costa Rica (PQ 1104), Atlantic salmon (Salmo salar) from Chile (PQ 1106) and three-line grunt (Parapristipoma trilineatum) from Japan (Ehime-1) were characterized by phenotypic and molecular taxonomic methods. These isolates were Gram-negative, oxidase negative, non-motile, strictly aerobic cocco-bacilli, produced H2S from cysteine supplemented media, which is phenotypically consistent with the genus Francisella. Comparison of 16S rRNA gene sequences and five partial housekeeping gene sequences (groEL, shdA, rpoB, rpoA and pgm) confirmed these isolates to be members of the genus Francisella, with high 16S rRNA similarity (> 99 %) to Francisella philomiragia subsp noatunensis, F. piscicida and Francisella philomiragia subsp philomiragia isolates. Despite the close 16s rRNA relationship with the aforementioned Francisella taxa, isolates PQ 1104 and Ehime-1 form a separate clade on phylogenetic analysis of the 16s rRNA gene and all housekeeping genes investigated, whereas isolate PQ 1106 is highly similar to F. philomiragia subsp noatunensis (NCIMB 14265T) and F. piscicida (DSM 18777T). DNA-DNA hybridization studies revealed mean reassociation values of 60.3 and 72.6 % between isolate PQ 1104 and F. philomiragia subsp noatunensis (NCIMB 14265T) and F. philomiragia subsp philomiragia (ATCC 25015), respectively. Thus, on the basis of molecular genetic evidence, we propose that isolates PQ 1104 and Ehime-1 should be recognised as Francisella asiatica sp. nov. with type strain PQ 1104T (NCIMB and CCUG number not received yet). No separation between F. piscicida and F. philomiragia subsp noatunensis were identified by the same methods and these species constitute heterotypic synonyms for which the epithet noatunensis has priority. However, given the increased evidence of ecological differentiation within the F. philomiragia group and the existence of a specific fish pathogenic clade, we propose that the F. philomiragia subsp noatunensis be elevated to species level as F. noatunensis comb. nov., sp.