Whole Genome Analysis of the Antibiotic Resistant Determinants and Mobile Elements Present in the Uncommon Pathogens Pseudomonas chlororaphis & Shewanella decolorationis

T LAZZARO; GISELA PARMECIANO DI NOTO; ANDRÉS IRIARTE; SABRINA MONTAÑA; S UONG; CECILIA QUIROGA; MARISA ALMUZARA; MARÍA SOLEDAD RAMÍREZ; CARLOS VAY

New Orleans

Congreso; ASM MICROBE 2017; 2017

Background: Implementation of new technologies in the diagnostic laboratory setting have contributed to the increase in identification of uncommon pathogens. These pathogens are often environmental microorganism with high level of resistance to several antibiotics transforming them into potential reservoirs of antibiotic resistance traits. Horizontal genetic transfer is a key mechanism in the development of antibiotic resistance. The aim of this project was to explore the whole genome sequence of two uncommon clinical isolates identified as Pseudomonas chlororaphis (Pc190) and Shewanella decolorationis (Shew256) in order to identify the presence of antibiotic resistant determinants and mobile elements that could be transfer to other bacterial species as well as to explain their multidrug resistant phenotypes.Methods: The draft genome sequence of Pc190 and Shew256 were obtained with Illumina MiSeq-I and Nextera XT DNA library. De novo assembly was performed with SPADES assembler 3.1.0 version. RAST was used to predict the open reading frames (ORF) and the predictions were confirmed using BLAST (version 2.0). Further genomic analysis was carried out using average nucleotide identity (ANI), ACT (Artemis), OrthoMCL, ARG-ANNOT, ISFinder, and PHAST, among others. PCR reactions and plasmid extraction were also performed.Results: The draft genomes of Pc190 and Shew256 consist of 6,791,658-bp and 5,365,006-bp sequences, respectively. By RAST server 6,052 and 4,766 protein-coding genes were predicted within the genomes of Pc190 and Shew256, respectively. The corresponding G+C contents were 63.0% and 47,9% for Pc190 and Shew256, respectively.Twelve resistant genes coding for resistance to seven different antibiotics families were present in Shew256, including β-lactamases such as PER-2. Several mobile elements were observed, such as insertion sequences, transposon, phages and the presence of an unusual class 1 integron. In Pc190 genome several antibiotic resistant gene werefound including β-lactamases, aminoglycoside modification enzymes and tetracycline resistant genes. A class 1 integron was present in addition to other mobile elements.Conclusion: A variety of resistance genes, which can explain the multidrug resistance phenotypes observed in both strains, as well as the presence of mobile elements, that can serve as tools in the mobilization of these determinants, were observed. We also highlight the importance of the isolation of uncommon multidrug resistance pathogens from clinical specimens.