| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1296-1 | ||||
Resumo:During a genomic surveillance study conducted to monitor the occurrence of clinically relevant antibiotic-resistant bacteria in urban ecosystems, a non-fermentative Gram-negative bacilli was isolates from the skin sample of a tree-frog Scinax fuscovarius inhabiting the campus of the University of São Paulo, Brazil. Strikingly, consecutive subcultures from a single pure colony originated two different colony sizes (i.e., > 2 and < 2 mm diameter). Both colonies were initially identified as Pseudomonas putida by VITEK 2 and MALDITOF-MS methods. The strains displayed resistance to cephalotin, meropenem and doripenem by using the disk-diffusion method with BrCAST interpretative criteria. On the other hand, while > 2 mm colonies exhibited a reduced inhibition halo for fosfomycin (strain 5FOST), < 2 mm colonies exhibited a reduced inhibition halo for rifampicin (strain 5RIF). Genomic DNA of 5FOST and 5RIF strains were extracted and sequenced by Illumina NextSeq system. In this regard, G+C content was 63.26 mol% for both isolates. Although, analysis of 16S rRNA gene of 5FOST and 5RIFstrains were 100% identical to each other, sharing >99.8% identity with P. putida DLL-E4T, P. inefficax JV551A3T, and P. shirazica VM14T, among others, the Neighbor-joining tree based on 16S rRNA gene sequences showed phylogenetic relationship between 5FOST, 5RIF, P. inefficax JV551A3T and P. ceruminis BML-PP028T clinical strains. Furthermore, phylogenetic analyses based on 16S rRNA, gyrB, rpoB, and rpoD housekeeping genes and MLSA confirms that 5FOST and 5RIF belong to the P. putida group. Genome BLAST distance phylogeny (GBDP) analysis using TYGS (https://tygs.dsmz.de/) revealed that 5FOST and 5RIF form a monophyletic clade with P. inefficax JV551A3T and P. asiatica RYU5T as their neighbors. ANIb, ANIm, dDDH, and TETRA genome-to-genome comparison values between 5FOST and 5RIF against Pseudomonas related species were ≤94.47, ≤57.4 and 0.9977%, respectively, which were below the thresholds limits for prokaryotic species delineation (i.e., 95, 70, and 0.9998%, respectively), confirming that 5FOST and 5RIF correspond to a new taxa for which the name Pseudomonas uspianensis sp. nov., and the type strain 5FOST (type strain deposits CCCT23.01T and IAL 9943T) are proposed. We characterized P. uspianensis as rod-shaped (1.9 x 3.8 µm), non-spore-forming and motile with one to five polar flagella. Fatty acid profiles of strains 5FOST and 5RIF displayed similarity index of 0.764 and 0.802, regarding to Pseudomonas-putida-biotype A and Pseudomonas sp., respectively. Finally, APIZYM revealed positive valine arylamidase and acid phosphatase that could distinguish these strains from other Pseudomonas species. In summary, we hereby report P. uspianensis sp. nov., a novel carbapenem-resistant species isolated from the tree-frog Scinax fuscovarius in São Paulo, Brazil. Palavras-chave: Pseudomonadaceae, genomic surveillance, Pseudomonas putida group, whole-genome sequence, novel species | |||||