tube Use of whole-genome sequencing Standard biochemical analyses : Comamonadaceae bacterium NML970112 JN585334 sugars (Table 1), API-ZYM and API 20E, to characterize zoonotically Comamonadaceae bacterium NML910035 JN585332 16s rRNA (Figure 1) and whole genome Comamonadaceae bacterium NML910036 JN585333 transmitted clinical isolates from the sequencing (WGS)(Table 2, 3) with 86 provisional taxon CDC Group NO-1 phylogenetic analyses were performed on all CDC Group NO-1 97BZ223 DQ054782 Comamonadaceae bacterium NML030146T JN585336 representing a new genus in the isolates. 100 Comamonadaceae bacterium NML120219 MF667936 family Comamonacaceae WGS - paired-end runs using the MiSeq 96 Comamonadaceae bacterium NML790751 JN585331 Sequencer (Illumina1.9, 2x 300 cycles) and 82 Comamonadaceae bacterium NML000135 JN585335 1 2 2 81 A-M. Bernier , T. Burdz , D. Wiebe , A. L. assembled and annotated using Spades Corticibacter populi 17B10-2-12T NR 137376 Pacheco 2, M. Unger 2, K. Bernard 2, 3 0.02 87 (version 3.5.1) and Prokka respectively. Xenophilus azovorans KF46FT NR 025114 1. U. de Saint-Boniface, Winnipeg, Canada 2. (Table 2). Lampropedia hyalina NBRC 102468T NR 114109 National Microbiology Lab., PHAC, Winnipeg, Canada 3. U of Manitoba, Winnipeg Canada. Comamonas terrigena IMI 359870T NR 028719 WGS comparison Pseudomonas aeruginosa DSM 50071T NR 117678 Introduction • JSpecies for Average Nucleotide Identity values (ANIb). Genome-to-genome Figure 1- 16S rRNA gene sequence alignment by Clustal W; relationship inferred by Neighbour-Joining, Mega Seven isolates from six patients collected over a distance calculator GGDC 2.1 for isDNA- 6.06; bootstrap value based on 1000 replicates. Scale represents substitutions per nucleotide position (2). 39 year period at the Canadian National DNA value (Table 3) Microbiology Laboratory (NML), recovered mostly • Single Nucleotide Variant PHYLogenomic NML (N=7) CDC (N=17) # of from human infections and associated with animal Strain Mb %GC contigs Coverage Origin • Feature frequency phylogeny (FFP) Gram negative, coccoid to 7/7 17/17 bites, were originally identified as CDC Group NML79-0751 3.18 66.63 55 87.8 cat bite wound analysis for phylogenic tree based on core medium rods, some vacuoles, NO-1 (1). This taxon group does not have formal features bipolar staining NML91-0035 3.30 65.29 34 58.7 dog bite wound taxonomic status and has never been genetically Colony: punctate, 0.25 mm(24h), 7/7 (100) 17/17 (100) characterized, making definitive assignment to NML91-0036 3.31 65.29 28 98.5 dog bite wound Proteome analysis by MALDI-TOF MS on a 0.5-1mm (48h), raised, convex, bacterial family, genus and species impossible. Bruker Microflex, (Bruker’s Biotyper 3.1 translucent [entire colony lifts NML97-0112 3.24 65.27 66 76.88 dog bite wound We performed a polyphasic taxonomic analysis database (7311 spectra)). (3) when touched with needle] NML00-0135 3.09 66.92 22 101.78 wound, right hand including whole genome sequencing of seven • No positive identification from Bruker Colony difficult to emulsify in 7/7(100) 17/17 (100) NML03-0146 3.08 65.44 13 66.1 peritoneal dialysate clinical sample isolates for the in the anticipation database, but in house MSPs could broth: of describing of a novel genus and species. identify each strain as one taxonomic NML120219 3.37 65.08 69 88 wound unknown Catalase + 7/7(100) 17/17 (100) group. Table 2- Assembly metrics and origins of the clinical strains (3) Oxidase + 1/7 (14) 0/17 (0) #ANIb top GGDC formula 2 bottom Bold = above cut off threshold (* within C.I.) Nitrate reduced 6/7 (86) 17/17 (100) Conclusion 1 2 3 4 5 6 7 8 MacConkey 1/7 NLF; 6/7 6/17 None of our isolates are unique in terms of biochemical NML00-0135 * 96.30 93.25 93.45 93.45 93.44 93.34 76.10 NG reactions, and while WGS analysis may indicate 2 NML79-0751 68.30* * 93.65 93.64 93.64 93.66 93.58 75.95 NOT REACTIVE: CHO or OF 7/7 (100) 17/17 (100) species, MALDI-TOF MS and 16S gene sequencing sugars; TSI; citrate; esculin; urea; cannot differentiate these isolates. Heuristically, NML03-0146 50.3 52.8 * 96.87 96.87 96.96 96.75 75.29 indol; gelatin; PPA; lysine; considering all biochemical and genetic data, we propose NML91-0035 51 52.9 73.7 * 100.00 99.63 96.63 75.39 arginine; ornithine; DNase; that the CDC Group NO-1 is a new genus and species in cetrimide; Na acetate; SS the family Comamonadaceae. NML91-0036 51 52.9 73.7 99 * 99.63 96.67 75.50 Ubiquinones Q8, minor ND NML97-0112 50.9 52.8 74.1 97.3 97.4 * 96.72 75.32 Q7 References (1) Hollis, D. G. et al. 1993. J. Clin. Microbiol. 31:746-748. NML120219 50.8 53.2 73.7 72.5 72.5 72.9 * 75.21 Table 1- Comparison of NML strains with the 17 strains (2) Bernard, K. A. et al. 2002b.. Anaerobe 8: 41–47. (3) Bernard, K.A. et al. 2016. IJSEM 66:4440-4444 L. hyalina 22.3 22.2 22.1 22.1 22.1 22 22 * described by the CDC (% in brackets) (1) Table 3- WGS comparisons using Genome to Genome distance calculator (formula 2, 70% cut Acknowledgements : we thank the Genome Core and the Proteomics Core of the NML for DNA Sequencing and MALDI-TOF MS off) and Anib (96-94% cut off) results using Jspecies (3)