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Kozyreva)] TJ ET 0.271 0.267 0.267 rg BT 115.657 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 121.079 613.351 Td /F1 9.8 Tf [(John Crandall)] TJ ET 0.271 0.267 0.267 rg BT 181.226 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 186.647 613.351 Td /F1 9.8 Tf [(Ashley Sabol)] TJ ET 0.271 0.267 0.267 rg BT 243.548 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 248.969 613.351 Td /F1 9.8 Tf [(Alyssa Poe)] TJ ET 0.271 0.267 0.267 rg BT 297.739 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 303.160 613.351 Td /F1 9.8 Tf [(Peng Zhang)] TJ ET 0.271 0.267 0.267 rg BT 356.278 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 361.699 613.351 Td /F1 9.8 Tf [(Jeniffer Concepcin-Acevedo)] TJ ET 0.271 0.267 0.267 rg BT 489.043 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 494.464 613.351 Td /F1 9.8 Tf [(Morgan N. )] TJ ET BT 26.250 601.446 Td /F1 9.8 Tf [(Schroeder)] TJ ET 0.271 0.267 0.267 rg BT 71.227 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 76.648 601.446 Td /F1 9.8 Tf [(Darlene Wagner)] TJ ET 0.271 0.267 0.267 rg BT 147.628 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 153.049 601.446 Td /F1 9.8 Tf [(Jeffrey Higa)] TJ ET 0.271 0.267 0.267 rg BT 205.065 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 210.486 601.446 Td /F1 9.8 Tf [(Eija Trees)] TJ ET 0.271 0.267 0.267 rg BT 254.371 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 259.792 601.446 Td /F1 9.8 Tf [(Vishnu Chaturvedi)] TJ ET 0.271 0.267 0.267 rg BT 26.250 589.542 Td /F1 9.8 Tf [(Kozyreva VK, Crandall J, Sabol A, Poe A, Zhang P, Concepcin-Acevedo J, Schroeder MN, Wagner D, Higa J, Trees E, )] TJ ET BT 26.250 577.637 Td /F1 9.8 Tf [(Chaturvedi V. Laboratory Investigation of )] TJ ET BT 205.631 577.637 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 291.245 577.637 Td /F1 9.8 Tf [( serovar Poona Outbreak in California: Comparison of Pulsed-)] TJ ET BT 26.250 565.732 Td /F1 9.8 Tf [(Field Gel Electrophoresis \(PFGE\) and Whole Genome Sequencing \(WGS\) Results. PLOS Currents Outbreaks. 2016 Nov 22 . )] TJ ET BT 26.250 553.827 Td /F1 9.8 Tf [(Edition 1. doi: 10.1371/currents.outbreaks.1bb3e36e74bd5779bc43ac3a8dae52e6.)] TJ ET q 15.000 33.443 577.500 518.003 re W n 0.271 0.267 0.267 rg BT 26.250 524.725 Td /F6 12.0 Tf [(Abstract)] TJ ET BT 26.250 504.771 Td /F1 9.8 Tf [(Introduction: Recently, )] TJ ET BT 125.963 504.771 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 214.288 504.771 Td /F1 9.8 Tf [(serovar Poona caused a multistate outbreak, with 245 out of 907 cases occurring in )] TJ ET BT 26.250 492.866 Td /F1 9.8 Tf [(California. We report a comparison of pulsed-field gel electrophoresis \(PFGE\) results with whole genome sequencing \(WGS\) for )] TJ ET BT 26.250 480.961 Td /F1 9.8 Tf [(genotyping of )] TJ ET BT 87.499 480.961 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 135.723 480.961 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET BT 26.250 461.556 Td /F1 9.8 Tf [(Methods: CA )] TJ ET BT 85.316 461.556 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 133.539 461.556 Td /F1 9.8 Tf [( Poona isolates, collected from July to August 2015, were genotyped by PFGE using XbaI restriction )] TJ ET BT 26.250 449.652 Td /F1 9.8 Tf [(enzyme. WGS was done using Nextera XT library kit with 2300 bp or 2250 bp sequencing chemistry on the Illumina MiSeq )] TJ ET BT 26.250 437.747 Td /F1 9.8 Tf [(Sequencer. Reads were mapped to the de novo assembled serovar Poona draft genome \(48 contigs, N50= 223,917\) from the )] TJ ET BT 26.250 425.842 Td /F1 9.8 Tf [(outbreak using CLCbio GW 8.0.2. The phylogenetic tree was generated based on hqSNPs calling. Genomes were annotated )] TJ ET BT 26.250 413.937 Td /F1 9.8 Tf [(with CGE and PHAST online tools. In silico MLST was performed using the CGE online tool.)] TJ ET BT 26.250 394.533 Td /F1 9.8 Tf [(Results: Human \(14\) and cucumber \(2\) )] TJ ET BT 197.470 394.533 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 245.693 394.533 Td /F1 9.8 Tf [( Poona isolates exhibited 3 possibly related PFGE patterns \(JL6X01.0018 )] TJ ET BT 26.250 382.628 Td /F1 9.8 Tf [([predominant], JL6X01.0375, JL6X01.0778\). All isolates that were related by PFGE also clustered together according to the )] TJ ET BT 26.250 370.723 Td /F1 9.8 Tf [(WGS. One isolate with a divergent PFGE pattern \(JL6X01.0776\) served as an outlier in the phylogenetic analysis and )] TJ ET BT 26.250 358.818 Td /F1 9.8 Tf [(substantially differed from the outbreak clade by WGS. All outbreak isolates were assigned to MLST sequence type 447. The )] TJ ET BT 26.250 346.914 Td /F1 9.8 Tf [(majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 318.341 346.914 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 366.564 346.914 Td /F1 9.8 Tf [( Pathogenicity Islands with few variations. One )] TJ ET BT 26.250 335.009 Td /F1 9.8 Tf [(outbreak isolate was sequenced and analyzed independently by CDC and CDPH laboratories; there was 0 SNP difference in )] TJ ET BT 26.250 323.104 Td /F1 9.8 Tf [(results. Additional two isolates were sequenced by CDC and the raw data was processed through CDPH and CDC analysis )] TJ ET BT 26.250 311.199 Td /F1 9.8 Tf [(pipelines. Both data analysis pipelines also generated concordant results.)] TJ ET BT 26.250 291.795 Td /F1 9.8 Tf [(Discussion: PFGE and WGS results for the recent CA )] TJ ET BT 260.874 291.795 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 349.199 291.795 Td /F1 9.8 Tf [(serovar Poona outbreak provided concordant )] TJ ET BT 26.250 279.890 Td /F1 9.8 Tf [(assignment of the isolates to the outbreak cluster. WGS allowed more robust determination of genetic relatedness, provided )] TJ ET BT 26.250 267.985 Td /F1 9.8 Tf [(information regarding MLST-type, pathogenicity genes, and bacteriophage content. WGS data obtained independently at two )] TJ ET BT 26.250 256.080 Td /F1 9.8 Tf [(laboratories showed complete agreement.)] TJ ET BT 26.250 219.478 Td /F6 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 199.524 Td /F1 9.8 Tf [(The authors received no specific funding for this work.)] TJ ET BT 26.250 170.421 Td /F6 12.0 Tf [(The Study)] TJ ET BT 26.250 150.467 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 111.865 150.467 Td /F1 9.8 Tf [( serotype Poona is a relatively rare serotype in the United States, responsible for about 1% of reported )] TJ ET BT 26.250 138.562 Td /F1 9.8 Tf [(human )] TJ ET BT 58.766 138.562 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 106.990 138.562 Td /F1 9.8 Tf [( cases)] TJ ET 0.267 0.267 0.267 rg BT 135.167 140.069 Td /F6 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 139.986 138.562 Td /F1 9.8 Tf [(. )] TJ ET BT 145.407 138.562 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 196.341 138.562 Td /F1 9.8 Tf [(Poona outbreaks are commonly attributed to farm-produced crops and goods)] TJ ET 0.267 0.267 0.267 rg BT 529.079 140.069 Td /F6 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 533.898 138.562 Td /F1 9.8 Tf [(. Serotype )] TJ ET BT 26.250 126.657 Td /F1 9.8 Tf [(Poona has also caused outbreaks associated with pet reptile exposure)] TJ ET 0.267 0.267 0.267 rg BT 330.820 128.165 Td /F6 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 335.639 126.657 Td /F1 9.8 Tf [(. Recently, )] TJ ET BT 384.409 126.657 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 432.632 126.657 Td /F1 9.8 Tf [( Poona caused a multistate )] TJ ET BT 26.250 114.753 Td /F1 9.8 Tf [(outbreak, with 245 out of 907 cases occurring in California. A multistate investigation identified imported cucumbers distributed )] TJ ET BT 26.250 102.848 Td /F1 9.8 Tf [(by Company A as the likely source of the outbreak)] TJ ET 0.267 0.267 0.267 rg BT 243.558 104.355 Td /F6 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 248.377 102.848 Td /F1 9.8 Tf [(. We report results of a real-time comparison of pulsed-field gel )] TJ ET BT 26.250 90.943 Td /F1 9.8 Tf [(electrophoresis \(PFGE\) and whole genome sequencing \(WGS\) for this )] TJ ET BT 332.946 90.943 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 381.170 90.943 Td /F1 9.8 Tf [( Poona outbreak investigation.)] TJ ET BT 26.250 71.538 Td /F1 9.8 Tf [(The )] TJ ET BT 45.760 71.538 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 93.983 71.538 Td /F1 9.8 Tf [( Poona isolates from California analyzed in this study are listed in Table 1. At the California Department of )] TJ ET BT 26.250 59.634 Td /F1 9.8 Tf [(Public Health \(CDPH\), PFGE was performed using )] TJ ET BT 248.394 59.634 Td /F5 9.8 Tf [(Xba)] TJ ET BT 265.739 59.634 Td /F1 9.8 Tf [(I macrorestriction per PulseNet USA protocol)] TJ ET 0.267 0.267 0.267 rg BT 459.716 61.141 Td /F6 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 464.534 59.634 Td /F1 9.8 Tf [(. For WGS, DNA libraries )] TJ ET BT 26.250 47.729 Td /F1 9.8 Tf [(were prepared using Nextera XT kit \(Illumina Inc., San Diego, CA\) and sequenced with 2300 bp or 2250 bp chemistry on the )] TJ ET Q q 15.000 634.458 577.500 103.542 re W n 0.267 0.267 0.267 rg BT 15.000 718.042 Td /F2 21.0 Tf [(Laboratory Investigation of)] TJ ET 0.271 0.267 0.267 rg BT 263.493 718.042 Td /F3 21.0 Tf [(Salmonella enterica)] TJ ET 0.267 0.267 0.267 rg BT 436.764 718.042 Td /F2 21.0 Tf [( serovar Poona )] TJ ET BT 15.000 693.094 Td /F2 21.0 Tf [(Outbreak in California: Comparison of Pulsed-Field Gel )] TJ ET BT 15.000 668.146 Td /F2 21.0 Tf [(Electrophoresis \(PFGE\) and Whole Genome Sequencing )] TJ ET BT 15.000 643.198 Td /F2 21.0 Tf [(\(WGS\) Results)] TJ ET Q 0.271 0.267 0.267 rg BT 15.000 625.192 Td /F4 9.8 Tf [(November 22, 2016)] TJ ET BT 96.510 625.192 Td /F4 9.8 Tf [()] TJ ET 0.267 0.267 0.267 rg BT 101.385 625.192 Td /F4 9.8 Tf [(Research Article)] TJ ET BT 26.250 613.351 Td /F1 9.8 Tf [(Varvara K. Kozyreva)] TJ ET 0.271 0.267 0.267 rg BT 115.657 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 121.079 613.351 Td /F1 9.8 Tf [(John Crandall)] TJ ET 0.271 0.267 0.267 rg BT 181.226 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 186.647 613.351 Td /F1 9.8 Tf [(Ashley Sabol)] TJ ET 0.271 0.267 0.267 rg BT 243.548 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 248.969 613.351 Td /F1 9.8 Tf [(Alyssa Poe)] TJ ET 0.271 0.267 0.267 rg BT 297.739 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 303.160 613.351 Td /F1 9.8 Tf [(Peng Zhang)] TJ ET 0.271 0.267 0.267 rg BT 356.278 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 361.699 613.351 Td /F1 9.8 Tf [(Jeniffer Concepcin-Acevedo)] TJ ET 0.271 0.267 0.267 rg BT 489.043 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 494.464 613.351 Td /F1 9.8 Tf [(Morgan N. )] TJ ET BT 26.250 601.446 Td /F1 9.8 Tf [(Schroeder)] TJ ET 0.271 0.267 0.267 rg BT 71.227 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 76.648 601.446 Td /F1 9.8 Tf [(Darlene Wagner)] TJ ET 0.271 0.267 0.267 rg BT 147.628 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 153.049 601.446 Td /F1 9.8 Tf [(Jeffrey Higa)] TJ ET 0.271 0.267 0.267 rg BT 205.065 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 210.486 601.446 Td /F1 9.8 Tf [(Eija Trees)] TJ ET 0.271 0.267 0.267 rg BT 254.371 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 259.792 601.446 Td /F1 9.8 Tf [(Vishnu Chaturvedi)] TJ ET 0.271 0.267 0.267 rg BT 26.250 589.542 Td /F1 9.8 Tf [(Kozyreva VK, Crandall J, Sabol A, Poe A, Zhang P, Concepcin-Acevedo J, Schroeder MN, Wagner D, Higa J, Trees E, )] TJ ET BT 26.250 577.637 Td /F1 9.8 Tf [(Chaturvedi V. Laboratory Investigation of )] TJ ET BT 205.631 577.637 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 291.245 577.637 Td /F1 9.8 Tf [( serovar Poona Outbreak in California: Comparison of Pulsed-)] TJ ET BT 26.250 565.732 Td /F1 9.8 Tf [(Field Gel Electrophoresis \(PFGE\) and Whole Genome Sequencing \(WGS\) Results. PLOS Currents Outbreaks. 2016 Nov 22 . )] TJ ET BT 26.250 553.827 Td /F1 9.8 Tf [(Edition 1. doi: 10.1371/currents.outbreaks.1bb3e36e74bd5779bc43ac3a8dae52e6.)] TJ ET q 15.000 33.443 577.500 518.003 re W n 0.271 0.267 0.267 rg BT 26.250 524.725 Td /F6 12.0 Tf [(Abstract)] TJ ET BT 26.250 504.771 Td /F1 9.8 Tf [(Introduction: Recently, )] TJ ET BT 125.963 504.771 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 214.288 504.771 Td /F1 9.8 Tf [(serovar Poona caused a multistate outbreak, with 245 out of 907 cases occurring in )] TJ ET BT 26.250 492.866 Td /F1 9.8 Tf [(California. We report a comparison of pulsed-field gel electrophoresis \(PFGE\) results with whole genome sequencing \(WGS\) for )] TJ ET BT 26.250 480.961 Td /F1 9.8 Tf [(genotyping of )] TJ ET BT 87.499 480.961 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 135.723 480.961 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET BT 26.250 461.556 Td /F1 9.8 Tf [(Methods: CA )] TJ ET BT 85.316 461.556 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 133.539 461.556 Td /F1 9.8 Tf [( Poona isolates, collected from July to August 2015, were genotyped by PFGE using XbaI restriction )] TJ ET BT 26.250 449.652 Td /F1 9.8 Tf [(enzyme. WGS was done using Nextera XT library kit with 2300 bp or 2250 bp sequencing chemistry on the Illumina MiSeq )] TJ ET BT 26.250 437.747 Td /F1 9.8 Tf [(Sequencer. Reads were mapped to the de novo assembled serovar Poona draft genome \(48 contigs, N50= 223,917\) from the )] TJ ET BT 26.250 425.842 Td /F1 9.8 Tf [(outbreak using CLCbio GW 8.0.2. The phylogenetic tree was generated based on hqSNPs calling. Genomes were annotated )] TJ ET BT 26.250 413.937 Td /F1 9.8 Tf [(with CGE and PHAST online tools. In silico MLST was performed using the CGE online tool.)] TJ ET BT 26.250 394.533 Td /F1 9.8 Tf [(Results: Human \(14\) and cucumber \(2\) )] TJ ET BT 197.470 394.533 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 245.693 394.533 Td /F1 9.8 Tf [( Poona isolates exhibited 3 possibly related PFGE patterns \(JL6X01.0018 )] TJ ET BT 26.250 382.628 Td /F1 9.8 Tf [([predominant], JL6X01.0375, JL6X01.0778\). All isolates that were related by PFGE also clustered together according to the )] TJ ET BT 26.250 370.723 Td /F1 9.8 Tf [(WGS. One isolate with a divergent PFGE pattern \(JL6X01.0776\) served as an outlier in the phylogenetic analysis and )] TJ ET BT 26.250 358.818 Td /F1 9.8 Tf [(substantially differed from the outbreak clade by WGS. All outbreak isolates were assigned to MLST sequence type 447. The )] TJ ET BT 26.250 346.914 Td /F1 9.8 Tf [(majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 318.341 346.914 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 366.564 346.914 Td /F1 9.8 Tf [( Pathogenicity Islands with few variations. One )] TJ ET BT 26.250 335.009 Td /F1 9.8 Tf [(outbreak isolate was sequenced and analyzed independently by CDC and CDPH laboratories; there was 0 SNP difference in )] TJ ET BT 26.250 323.104 Td /F1 9.8 Tf [(results. Additional two isolates were sequenced by CDC and the raw data was processed through CDPH and CDC analysis )] TJ ET BT 26.250 311.199 Td /F1 9.8 Tf [(pipelines. Both data analysis pipelines also generated concordant results.)] TJ ET BT 26.250 291.795 Td /F1 9.8 Tf [(Discussion: PFGE and WGS results for the recent CA )] TJ ET BT 260.874 291.795 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 349.199 291.795 Td /F1 9.8 Tf [(serovar Poona outbreak provided concordant )] TJ ET BT 26.250 279.890 Td /F1 9.8 Tf [(assignment of the isolates to the outbreak cluster. WGS allowed more robust determination of genetic relatedness, provided )] TJ ET BT 26.250 267.985 Td /F1 9.8 Tf [(information regarding MLST-type, pathogenicity genes, and bacteriophage content. WGS data obtained independently at two )] TJ ET BT 26.250 256.080 Td /F1 9.8 Tf [(laboratories showed complete agreement.)] TJ ET BT 26.250 219.478 Td /F6 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 199.524 Td /F1 9.8 Tf [(The authors received no specific funding for this work.)] TJ ET BT 26.250 170.421 Td /F6 12.0 Tf [(The Study)] TJ ET BT 26.250 150.467 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 111.865 150.467 Td /F1 9.8 Tf [( serotype Poona is a relatively rare serotype in the United States, responsible for about 1% of reported )] TJ ET BT 26.250 138.562 Td /F1 9.8 Tf [(human )] TJ ET BT 58.766 138.562 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 106.990 138.562 Td /F1 9.8 Tf [( cases)] TJ ET 0.267 0.267 0.267 rg BT 135.167 140.069 Td /F6 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 139.986 138.562 Td /F1 9.8 Tf [(. )] TJ ET BT 145.407 138.562 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 196.341 138.562 Td /F1 9.8 Tf [(Poona outbreaks are commonly attributed to farm-produced crops and goods)] TJ ET 0.267 0.267 0.267 rg BT 529.079 140.069 Td /F6 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 533.898 138.562 Td /F1 9.8 Tf [(. Serotype )] TJ ET BT 26.250 126.657 Td /F1 9.8 Tf [(Poona has also caused outbreaks associated with pet reptile exposure)] TJ ET 0.267 0.267 0.267 rg BT 330.820 128.165 Td /F6 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 335.639 126.657 Td /F1 9.8 Tf [(. Recently, )] TJ ET BT 384.409 126.657 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 432.632 126.657 Td /F1 9.8 Tf [( Poona caused a multistate )] TJ ET BT 26.250 114.753 Td /F1 9.8 Tf [(outbreak, with 245 out of 907 cases occurring in California. A multistate investigation identified imported cucumbers distributed )] TJ ET BT 26.250 102.848 Td /F1 9.8 Tf [(by Company A as the likely source of the outbreak)] TJ ET 0.267 0.267 0.267 rg BT 243.558 104.355 Td /F6 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 248.377 102.848 Td /F1 9.8 Tf [(. We report results of a real-time comparison of pulsed-field gel )] TJ ET BT 26.250 90.943 Td /F1 9.8 Tf [(electrophoresis \(PFGE\) and whole genome sequencing \(WGS\) for this )] TJ ET BT 332.946 90.943 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 381.170 90.943 Td /F1 9.8 Tf [( Poona outbreak investigation.)] TJ ET BT 26.250 71.538 Td /F1 9.8 Tf [(The )] TJ ET BT 45.760 71.538 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 93.983 71.538 Td /F1 9.8 Tf [( Poona isolates from California analyzed in this study are listed in Table 1. At the California Department of )] TJ ET BT 26.250 59.634 Td /F1 9.8 Tf [(Public Health \(CDPH\), PFGE was performed using )] TJ ET BT 248.394 59.634 Td /F5 9.8 Tf [(Xba)] TJ ET BT 265.739 59.634 Td /F1 9.8 Tf [(I macrorestriction per PulseNet USA protocol)] TJ ET 0.267 0.267 0.267 rg BT 459.716 61.141 Td /F6 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 464.534 59.634 Td /F1 9.8 Tf [(. For WGS, DNA libraries )] TJ ET BT 26.250 47.729 Td /F1 9.8 Tf [(were prepared using Nextera XT kit \(Illumina Inc., San Diego, CA\) and sequenced with 2300 bp or 2250 bp chemistry on the )] TJ ET Q q 15.000 634.458 577.500 103.542 re W n 0.267 0.267 0.267 rg BT 15.000 718.042 Td /F2 21.0 Tf [(Laboratory Investigation of)] TJ ET 0.271 0.267 0.267 rg BT 263.493 718.042 Td /F3 21.0 Tf [(Salmonella enterica)] TJ ET 0.267 0.267 0.267 rg BT 436.764 718.042 Td /F2 21.0 Tf [( serovar Poona )] TJ ET BT 15.000 693.094 Td /F2 21.0 Tf [(Outbreak in California: Comparison of Pulsed-Field Gel )] TJ ET BT 15.000 668.146 Td /F2 21.0 Tf [(Electrophoresis \(PFGE\) and Whole Genome Sequencing )] TJ ET BT 15.000 643.198 Td /F2 21.0 Tf [(\(WGS\) Results)] TJ ET Q 0.271 0.267 0.267 rg BT 15.000 625.192 Td /F4 9.8 Tf [(November 22, 2016)] TJ ET BT 96.510 625.192 Td /F4 9.8 Tf [()] TJ ET 0.267 0.267 0.267 rg BT 101.385 625.192 Td /F4 9.8 Tf [(Research Article)] TJ ET BT 26.250 613.351 Td /F1 9.8 Tf [(Varvara K. Kozyreva)] TJ ET 0.271 0.267 0.267 rg BT 115.657 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 121.079 613.351 Td /F1 9.8 Tf [(John Crandall)] TJ ET 0.271 0.267 0.267 rg BT 181.226 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 186.647 613.351 Td /F1 9.8 Tf [(Ashley Sabol)] TJ ET 0.271 0.267 0.267 rg BT 243.548 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 248.969 613.351 Td /F1 9.8 Tf [(Alyssa Poe)] TJ ET 0.271 0.267 0.267 rg BT 297.739 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 303.160 613.351 Td /F1 9.8 Tf [(Peng Zhang)] TJ ET 0.271 0.267 0.267 rg BT 356.278 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 361.699 613.351 Td /F1 9.8 Tf [(Jeniffer Concepcin-Acevedo)] TJ ET 0.271 0.267 0.267 rg BT 489.043 613.351 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 494.464 613.351 Td /F1 9.8 Tf [(Morgan N. )] TJ ET BT 26.250 601.446 Td /F1 9.8 Tf [(Schroeder)] TJ ET 0.271 0.267 0.267 rg BT 71.227 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 76.648 601.446 Td /F1 9.8 Tf [(Darlene Wagner)] TJ ET 0.271 0.267 0.267 rg BT 147.628 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 153.049 601.446 Td /F1 9.8 Tf [(Jeffrey Higa)] TJ ET 0.271 0.267 0.267 rg BT 205.065 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 210.486 601.446 Td /F1 9.8 Tf [(Eija Trees)] TJ ET 0.271 0.267 0.267 rg BT 254.371 601.446 Td /F1 9.8 Tf [(, )] TJ ET 0.267 0.267 0.267 rg BT 259.792 601.446 Td /F1 9.8 Tf [(Vishnu Chaturvedi)] TJ ET 0.271 0.267 0.267 rg BT 26.250 589.542 Td /F1 9.8 Tf [(Kozyreva VK, Crandall J, Sabol A, Poe A, Zhang P, Concepcin-Acevedo J, Schroeder MN, Wagner D, Higa J, Trees E, )] TJ ET BT 26.250 577.637 Td /F1 9.8 Tf [(Chaturvedi V. Laboratory Investigation of )] TJ ET BT 205.631 577.637 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 291.245 577.637 Td /F1 9.8 Tf [( serovar Poona Outbreak in California: Comparison of Pulsed-)] TJ ET BT 26.250 565.732 Td /F1 9.8 Tf [(Field Gel Electrophoresis \(PFGE\) and Whole Genome Sequencing \(WGS\) Results. PLOS Currents Outbreaks. 2016 Nov 22 . )] TJ ET BT 26.250 553.827 Td /F1 9.8 Tf [(Edition 1. doi: 10.1371/currents.outbreaks.1bb3e36e74bd5779bc43ac3a8dae52e6.)] TJ ET q 15.000 33.443 577.500 518.003 re W n 0.271 0.267 0.267 rg BT 26.250 524.725 Td /F6 12.0 Tf [(Abstract)] TJ ET BT 26.250 504.771 Td /F1 9.8 Tf [(Introduction: Recently, )] TJ ET BT 125.963 504.771 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 214.288 504.771 Td /F1 9.8 Tf [(serovar Poona caused a multistate outbreak, with 245 out of 907 cases occurring in )] TJ ET BT 26.250 492.866 Td /F1 9.8 Tf [(California. We report a comparison of pulsed-field gel electrophoresis \(PFGE\) results with whole genome sequencing \(WGS\) for )] TJ ET BT 26.250 480.961 Td /F1 9.8 Tf [(genotyping of )] TJ ET BT 87.499 480.961 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 135.723 480.961 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET BT 26.250 461.556 Td /F1 9.8 Tf [(Methods: CA )] TJ ET BT 85.316 461.556 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 133.539 461.556 Td /F1 9.8 Tf [( Poona isolates, collected from July to August 2015, were genotyped by PFGE using XbaI restriction )] TJ ET BT 26.250 449.652 Td /F1 9.8 Tf [(enzyme. WGS was done using Nextera XT library kit with 2300 bp or 2250 bp sequencing chemistry on the Illumina MiSeq )] TJ ET BT 26.250 437.747 Td /F1 9.8 Tf [(Sequencer. Reads were mapped to the de novo assembled serovar Poona draft genome \(48 contigs, N50= 223,917\) from the )] TJ ET BT 26.250 425.842 Td /F1 9.8 Tf [(outbreak using CLCbio GW 8.0.2. The phylogenetic tree was generated based on hqSNPs calling. Genomes were annotated )] TJ ET BT 26.250 413.937 Td /F1 9.8 Tf [(with CGE and PHAST online tools. In silico MLST was performed using the CGE online tool.)] TJ ET BT 26.250 394.533 Td /F1 9.8 Tf [(Results: Human \(14\) and cucumber \(2\) )] TJ ET BT 197.470 394.533 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 245.693 394.533 Td /F1 9.8 Tf [( Poona isolates exhibited 3 possibly related PFGE patterns \(JL6X01.0018 )] TJ ET BT 26.250 382.628 Td /F1 9.8 Tf [([predominant], JL6X01.0375, JL6X01.0778\). All isolates that were related by PFGE also clustered together according to the )] TJ ET BT 26.250 370.723 Td /F1 9.8 Tf [(WGS. One isolate with a divergent PFGE pattern \(JL6X01.0776\) served as an outlier in the phylogenetic analysis and )] TJ ET BT 26.250 358.818 Td /F1 9.8 Tf [(substantially differed from the outbreak clade by WGS. All outbreak isolates were assigned to MLST sequence type 447. The )] TJ ET BT 26.250 346.914 Td /F1 9.8 Tf [(majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 318.341 346.914 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 366.564 346.914 Td /F1 9.8 Tf [( Pathogenicity Islands with few variations. One )] TJ ET BT 26.250 335.009 Td /F1 9.8 Tf [(outbreak isolate was sequenced and analyzed independently by CDC and CDPH laboratories; there was 0 SNP difference in )] TJ ET BT 26.250 323.104 Td /F1 9.8 Tf [(results. Additional two isolates were sequenced by CDC and the raw data was processed through CDPH and CDC analysis )] TJ ET BT 26.250 311.199 Td /F1 9.8 Tf [(pipelines. Both data analysis pipelines also generated concordant results.)] TJ ET BT 26.250 291.795 Td /F1 9.8 Tf [(Discussion: PFGE and WGS results for the recent CA )] TJ ET BT 260.874 291.795 Td /F5 9.8 Tf [(Salmonella enterica )] TJ ET BT 349.199 291.795 Td /F1 9.8 Tf [(serovar Poona outbreak provided concordant )] TJ ET BT 26.250 279.890 Td /F1 9.8 Tf [(assignment of the isolates to the outbreak cluster. WGS allowed more robust determination of genetic relatedness, provided )] TJ ET BT 26.250 267.985 Td /F1 9.8 Tf [(information regarding MLST-type, pathogenicity genes, and bacteriophage content. WGS data obtained independently at two )] TJ ET BT 26.250 256.080 Td /F1 9.8 Tf [(laboratories showed complete agreement.)] TJ ET BT 26.250 219.478 Td /F6 12.0 Tf [(Funding Statement)] TJ ET BT 26.250 199.524 Td /F1 9.8 Tf [(The authors received no specific funding for this work.)] TJ ET BT 26.250 170.421 Td /F6 12.0 Tf [(The Study)] TJ ET BT 26.250 150.467 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 111.865 150.467 Td /F1 9.8 Tf [( serotype Poona is a relatively rare serotype in the United States, responsible for about 1% of reported )] TJ ET BT 26.250 138.562 Td /F1 9.8 Tf [(human )] TJ ET BT 58.766 138.562 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 106.990 138.562 Td /F1 9.8 Tf [( cases)] TJ ET 0.267 0.267 0.267 rg BT 135.167 140.069 Td /F6 8.7 Tf [(1)] TJ ET 0.271 0.267 0.267 rg BT 139.986 138.562 Td /F1 9.8 Tf [(. )] TJ ET BT 145.407 138.562 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 196.341 138.562 Td /F1 9.8 Tf [(Poona outbreaks are commonly attributed to farm-produced crops and goods)] TJ ET 0.267 0.267 0.267 rg BT 529.079 140.069 Td /F6 8.7 Tf [(2)] TJ ET 0.271 0.267 0.267 rg BT 533.898 138.562 Td /F1 9.8 Tf [(. Serotype )] TJ ET BT 26.250 126.657 Td /F1 9.8 Tf [(Poona has also caused outbreaks associated with pet reptile exposure)] TJ ET 0.267 0.267 0.267 rg BT 330.820 128.165 Td /F6 8.7 Tf [(3)] TJ ET 0.271 0.267 0.267 rg BT 335.639 126.657 Td /F1 9.8 Tf [(. Recently, )] TJ ET BT 384.409 126.657 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 432.632 126.657 Td /F1 9.8 Tf [( Poona caused a multistate )] TJ ET BT 26.250 114.753 Td /F1 9.8 Tf [(outbreak, with 245 out of 907 cases occurring in California. A multistate investigation identified imported cucumbers distributed )] TJ ET BT 26.250 102.848 Td /F1 9.8 Tf [(by Company A as the likely source of the outbreak)] TJ ET 0.267 0.267 0.267 rg BT 243.558 104.355 Td /F6 8.7 Tf [(4)] TJ ET 0.271 0.267 0.267 rg BT 248.377 102.848 Td /F1 9.8 Tf [(. We report results of a real-time comparison of pulsed-field gel )] TJ ET BT 26.250 90.943 Td /F1 9.8 Tf [(electrophoresis \(PFGE\) and whole genome sequencing \(WGS\) for this )] TJ ET BT 332.946 90.943 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 381.170 90.943 Td /F1 9.8 Tf [( Poona outbreak investigation.)] TJ ET BT 26.250 71.538 Td /F1 9.8 Tf [(The )] TJ ET BT 45.760 71.538 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 93.983 71.538 Td /F1 9.8 Tf [( Poona isolates from California analyzed in this study are listed in Table 1. At the California Department of )] TJ ET BT 26.250 59.634 Td /F1 9.8 Tf [(Public Health \(CDPH\), PFGE was performed using )] TJ ET BT 248.394 59.634 Td /F5 9.8 Tf [(Xba)] TJ ET BT 265.739 59.634 Td /F1 9.8 Tf [(I macrorestriction per PulseNet USA protocol)] TJ ET 0.267 0.267 0.267 rg BT 459.716 61.141 Td /F6 8.7 Tf [(5)] TJ ET 0.271 0.267 0.267 rg BT 464.534 59.634 Td /F1 9.8 Tf [(. For WGS, DNA libraries )] TJ ET BT 26.250 47.729 Td /F1 9.8 Tf [(were prepared using Nextera XT kit \(Illumina Inc., San Diego, CA\) and sequenced with 2300 bp or 2250 bp chemistry on the )] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(1)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 8 0 obj << /Type /Font /Subtype /Type1 /Name /F1 /BaseFont /Helvetica /Encoding /WinAnsiEncoding >> endobj 9 0 obj << /Type /Font /Subtype /Type1 /Name /F2 /BaseFont /Times-Bold /Encoding /WinAnsiEncoding >> endobj 10 0 obj << /Type /Font /Subtype /Type1 /Name /F3 /BaseFont /Times-BoldItalic /Encoding /WinAnsiEncoding >> endobj 11 0 obj << /Type /Font /Subtype /Type1 /Name /F4 /BaseFont /Times-Italic /Encoding /WinAnsiEncoding >> endobj 12 0 obj << /Type /Font /Subtype /Type1 /Name /F5 /BaseFont /Helvetica-Oblique /Encoding /WinAnsiEncoding >> endobj 13 0 obj << /Type /Font /Subtype /Type1 /Name /F6 /BaseFont /Helvetica-Bold /Encoding 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The analysis of sequencing data at CDPH was performed as follows:)] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Genomes were annotated with prokka v1.1 tool)] TJ ET 0.267 0.267 0.267 rg BT 230.015 737.674 Td /F6 8.7 Tf [(6)] TJ ET 0.271 0.267 0.267 rg BT 234.834 736.167 Td /F1 9.8 Tf [(, the Center for Genomic Epidemiology \(CGE\), and Phage Search Tool )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(\(PHAST\) online resources)] TJ ET 0.267 0.267 0.267 rg BT 139.487 725.769 Td /F6 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 144.305 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 146.715 725.769 Td /F6 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 151.533 724.262 Td /F1 9.8 Tf [(. )] TJ ET BT 156.954 724.262 Td /F5 9.8 Tf [(In silico)] TJ ET BT 189.461 724.262 Td /F1 9.8 Tf [( multi-locus sequence typing \(MLST\) was performed using the CGE online tool)] TJ ET 0.267 0.267 0.267 rg BT 528.127 725.769 Td /F6 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 532.946 724.262 Td /F1 9.8 Tf [( against )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(the 7-gene MLST database of University of Warwick \(http://mlst.warwick.ac.uk/mlst/dbs/Senterica\). The )] TJ ET BT 473.814 712.357 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(Pathogenicity Islands \(SPI\) were characterized using SPIFinder 1.0 CGE tool \(https://cge.cbs.dtu.dk/services/SPIFinder/\).)] TJ ET BT 26.250 681.048 Td /F1 9.8 Tf [(Paired-end reads were mapped to the )] TJ ET BT 192.624 681.048 Td /F5 9.8 Tf [(de novo)] TJ ET BT 227.315 681.048 Td /F1 9.8 Tf [( assembled outbreak )] TJ ET BT 320.524 681.048 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 368.748 681.048 Td /F1 9.8 Tf [( Poona M15X04725 draft genome \(48 contigs, )] TJ ET BT 26.250 669.143 Td /F1 9.8 Tf [(N50= 223,917\) using CLCbio Genomic Workbench 8.0.2 \(Qiagen, Aarhus, Denmark\). Genes of mobile and prophage elements )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(in the reference )] TJ ET BT 96.704 657.238 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 144.927 657.238 Td /F1 9.8 Tf [( Poona M15X04725 genome were annotated using prokka v1.1 and PHAST tools and consequently )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(masked from mapping in CLCbio Genomic Workbench 8.0.2. SNPs were then called in coding and non-coding genome areas )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(using SAMtools mpileup \(v.1.2;)] TJ ET 0.267 0.267 0.267 rg BT 160.634 634.936 Td /F6 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 170.272 633.429 Td /F1 9.8 Tf [(\) and converted into VCF matrix using bcftools \(v0.1.19; http://samtools.github.io/bcftools/\). )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(Variants were parsed using vcftools \(v.0.1.12b;)] TJ ET 0.267 0.267 0.267 rg BT 228.923 623.031 Td /F6 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 238.561 621.524 Td /F1 9.8 Tf [(\) to include only high-quality SNPs \(hqSNPs\) with minimum position coverage )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(?30x and minimum quality of the base > 200 \(min-meanDP 30; minQ 200\), with InDels and the heterozygote calls excluded. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(The Maximum Likelihood phylogenetic tree was generated based on hqSNPs calls under the Jukes-Cantor nucleotide )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(substitution model; with 100 bootstrap replicates.)] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(At the US Centers for Disease Control and Prevention \(CDC\) LyveSET 1.1.4 SNP calling pipeline )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(\(http://www.github.com/lskatz/lyve-SET\) was applied with reads trimmed using fastx_trimmer 5 bases from 5 ends before )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(mapping single-end by SMALT. SNPs were called using Varscan at > 20x coverage, > 95% read support, and ?5 bp apart. The )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(draft assembly of FDA00009409 from the outbreak \(38 contigs\) was used as a reference without prophage masking.)] TJ ET BT 26.250 511.286 Td /F1 9.8 Tf [(Fourteen human and two cucumber )] TJ ET BT 182.864 511.286 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 231.088 511.286 Td /F1 9.8 Tf [( Poona isolates analyzed in this study exhibited 3 possibly related PFGE )] TJ ET BT 26.250 499.381 Td /F1 9.8 Tf [(patterns \(JL6X01.0018 [predominant], JL6X01.0375, JL6X01.0778\). The isolates were selected to include representatives of the )] TJ ET BT 26.250 487.476 Td /F1 9.8 Tf [(three above mentioned PFGE patterns and epidemiologically representative isolates with and without links to the cucumber )] TJ ET BT 26.250 475.572 Td /F1 9.8 Tf [(source. In the interest of the cost effectiveness, the WGS was limited to 16 isolates since the detected PFGE patterns were rare )] TJ ET BT 26.250 463.667 Td /F1 9.8 Tf [(in California and only few SNP differences were found among the sampled isolates. Of the 14 case-patients whose isolates )] TJ ET BT 26.250 451.762 Td /F1 9.8 Tf [(were analyzed, 10 \(71%\) reported consuming cucumbers during the week preceding their illness onsets, 2 case-patients denied )] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(consuming any cucumbers, and cucumber exposure was unknown for 2 case-patients \(Table 1\). The cucumber exposures )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(reported by the 10 case-patients were all linked to Company A. One isolate \(M15X03586\) with a divergent PFGE pattern )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(\(JL6X01.0776\) served as an outlier in the phylogenetic analysis \(Table 1, Figure 1A\). Cucumber exposure for the patient was )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(unknown; the patient was not considered part of the outbreak cluster based on PFGE.)] TJ ET 0.965 0.965 0.965 rg 26.250 59.298 555.000 334.964 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 394.262 m 581.250 394.262 l 581.250 393.512 l 26.250 393.512 l f q 450.000 0 0 277.500 35.250 107.012 cm /I3 Do Q q 35.250 59.298 537.000 41.714 re W n Q Q q 15.000 59.298 577.500 717.702 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(Illumina MiSeq at 49-124x sequencing coverage \(see NCBI accession numbers for the corresponding sequences in Table 1\). )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(Time-to-results was 4 days for PFGE and 7 days for WGS. The analysis of sequencing data at CDPH was performed as follows:)] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Genomes were annotated with prokka v1.1 tool)] TJ ET 0.267 0.267 0.267 rg BT 230.015 737.674 Td /F6 8.7 Tf [(6)] TJ ET 0.271 0.267 0.267 rg BT 234.834 736.167 Td /F1 9.8 Tf [(, the Center for Genomic Epidemiology \(CGE\), and Phage Search Tool )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(\(PHAST\) online resources)] TJ ET 0.267 0.267 0.267 rg BT 139.487 725.769 Td /F6 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 144.305 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 146.715 725.769 Td /F6 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 151.533 724.262 Td /F1 9.8 Tf [(. )] TJ ET BT 156.954 724.262 Td /F5 9.8 Tf [(In silico)] TJ ET BT 189.461 724.262 Td /F1 9.8 Tf [( multi-locus sequence typing \(MLST\) was performed using the CGE online tool)] TJ ET 0.267 0.267 0.267 rg BT 528.127 725.769 Td /F6 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 532.946 724.262 Td /F1 9.8 Tf [( against )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(the 7-gene MLST database of University of Warwick \(http://mlst.warwick.ac.uk/mlst/dbs/Senterica\). The )] TJ ET BT 473.814 712.357 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(Pathogenicity Islands \(SPI\) were characterized using SPIFinder 1.0 CGE tool \(https://cge.cbs.dtu.dk/services/SPIFinder/\).)] TJ ET BT 26.250 681.048 Td /F1 9.8 Tf [(Paired-end reads were mapped to the )] TJ ET BT 192.624 681.048 Td /F5 9.8 Tf [(de novo)] TJ ET BT 227.315 681.048 Td /F1 9.8 Tf [( assembled outbreak )] TJ ET BT 320.524 681.048 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 368.748 681.048 Td /F1 9.8 Tf [( Poona M15X04725 draft genome \(48 contigs, )] TJ ET BT 26.250 669.143 Td /F1 9.8 Tf [(N50= 223,917\) using CLCbio Genomic Workbench 8.0.2 \(Qiagen, Aarhus, Denmark\). Genes of mobile and prophage elements )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(in the reference )] TJ ET BT 96.704 657.238 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 144.927 657.238 Td /F1 9.8 Tf [( Poona M15X04725 genome were annotated using prokka v1.1 and PHAST tools and consequently )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(masked from mapping in CLCbio Genomic Workbench 8.0.2. SNPs were then called in coding and non-coding genome areas )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(using SAMtools mpileup \(v.1.2;)] TJ ET 0.267 0.267 0.267 rg BT 160.634 634.936 Td /F6 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 170.272 633.429 Td /F1 9.8 Tf [(\) and converted into VCF matrix using bcftools \(v0.1.19; http://samtools.github.io/bcftools/\). )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(Variants were parsed using vcftools \(v.0.1.12b;)] TJ ET 0.267 0.267 0.267 rg BT 228.923 623.031 Td /F6 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 238.561 621.524 Td /F1 9.8 Tf [(\) to include only high-quality SNPs \(hqSNPs\) with minimum position coverage )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(?30x and minimum quality of the base > 200 \(min-meanDP 30; minQ 200\), with InDels and the heterozygote calls excluded. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(The Maximum Likelihood phylogenetic tree was generated based on hqSNPs calls under the Jukes-Cantor nucleotide )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(substitution model; with 100 bootstrap replicates.)] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(At the US Centers for Disease Control and Prevention \(CDC\) LyveSET 1.1.4 SNP calling pipeline )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(\(http://www.github.com/lskatz/lyve-SET\) was applied with reads trimmed using fastx_trimmer 5 bases from 5 ends before )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(mapping single-end by SMALT. SNPs were called using Varscan at > 20x coverage, > 95% read support, and ?5 bp apart. The )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(draft assembly of FDA00009409 from the outbreak \(38 contigs\) was used as a reference without prophage masking.)] TJ ET BT 26.250 511.286 Td /F1 9.8 Tf [(Fourteen human and two cucumber )] TJ ET BT 182.864 511.286 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 231.088 511.286 Td /F1 9.8 Tf [( Poona isolates analyzed in this study exhibited 3 possibly related PFGE )] TJ ET BT 26.250 499.381 Td /F1 9.8 Tf [(patterns \(JL6X01.0018 [predominant], JL6X01.0375, JL6X01.0778\). The isolates were selected to include representatives of the )] TJ ET BT 26.250 487.476 Td /F1 9.8 Tf [(three above mentioned PFGE patterns and epidemiologically representative isolates with and without links to the cucumber )] TJ ET BT 26.250 475.572 Td /F1 9.8 Tf [(source. In the interest of the cost effectiveness, the WGS was limited to 16 isolates since the detected PFGE patterns were rare )] TJ ET BT 26.250 463.667 Td /F1 9.8 Tf [(in California and only few SNP differences were found among the sampled isolates. Of the 14 case-patients whose isolates )] TJ ET BT 26.250 451.762 Td /F1 9.8 Tf [(were analyzed, 10 \(71%\) reported consuming cucumbers during the week preceding their illness onsets, 2 case-patients denied )] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(consuming any cucumbers, and cucumber exposure was unknown for 2 case-patients \(Table 1\). The cucumber exposures )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(reported by the 10 case-patients were all linked to Company A. One isolate \(M15X03586\) with a divergent PFGE pattern )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(\(JL6X01.0776\) served as an outlier in the phylogenetic analysis \(Table 1, Figure 1A\). Cucumber exposure for the patient was )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(unknown; the patient was not considered part of the outbreak cluster based on PFGE.)] TJ ET 0.965 0.965 0.965 rg 26.250 59.298 555.000 334.964 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 394.262 m 581.250 394.262 l 581.250 393.512 l 26.250 393.512 l f q 450.000 0 0 277.500 35.250 107.012 cm /I3 Do Q q 35.250 59.298 537.000 41.714 re W n Q Q q 15.000 59.298 577.500 717.702 re W n 0.271 0.267 0.267 rg BT 26.250 767.476 Td /F1 9.8 Tf [(Illumina MiSeq at 49-124x sequencing coverage \(see NCBI accession numbers for the corresponding sequences in Table 1\). )] TJ ET BT 26.250 755.571 Td /F1 9.8 Tf [(Time-to-results was 4 days for PFGE and 7 days for WGS. The analysis of sequencing data at CDPH was performed as follows:)] TJ ET BT 26.250 736.167 Td /F1 9.8 Tf [(Genomes were annotated with prokka v1.1 tool)] TJ ET 0.267 0.267 0.267 rg BT 230.015 737.674 Td /F6 8.7 Tf [(6)] TJ ET 0.271 0.267 0.267 rg BT 234.834 736.167 Td /F1 9.8 Tf [(, the Center for Genomic Epidemiology \(CGE\), and Phage Search Tool )] TJ ET BT 26.250 724.262 Td /F1 9.8 Tf [(\(PHAST\) online resources)] TJ ET 0.267 0.267 0.267 rg BT 139.487 725.769 Td /F6 8.7 Tf [(7)] TJ ET 0.271 0.267 0.267 rg BT 144.305 728.150 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 146.715 725.769 Td /F6 8.7 Tf [(8)] TJ ET 0.271 0.267 0.267 rg BT 151.533 724.262 Td /F1 9.8 Tf [(. )] TJ ET BT 156.954 724.262 Td /F5 9.8 Tf [(In silico)] TJ ET BT 189.461 724.262 Td /F1 9.8 Tf [( multi-locus sequence typing \(MLST\) was performed using the CGE online tool)] TJ ET 0.267 0.267 0.267 rg BT 528.127 725.769 Td /F6 8.7 Tf [(9)] TJ ET 0.271 0.267 0.267 rg BT 532.946 724.262 Td /F1 9.8 Tf [( against )] TJ ET BT 26.250 712.357 Td /F1 9.8 Tf [(the 7-gene MLST database of University of Warwick \(http://mlst.warwick.ac.uk/mlst/dbs/Senterica\). The )] TJ ET BT 473.814 712.357 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 26.250 700.452 Td /F1 9.8 Tf [(Pathogenicity Islands \(SPI\) were characterized using SPIFinder 1.0 CGE tool \(https://cge.cbs.dtu.dk/services/SPIFinder/\).)] TJ ET BT 26.250 681.048 Td /F1 9.8 Tf [(Paired-end reads were mapped to the )] TJ ET BT 192.624 681.048 Td /F5 9.8 Tf [(de novo)] TJ ET BT 227.315 681.048 Td /F1 9.8 Tf [( assembled outbreak )] TJ ET BT 320.524 681.048 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 368.748 681.048 Td /F1 9.8 Tf [( Poona M15X04725 draft genome \(48 contigs, )] TJ ET BT 26.250 669.143 Td /F1 9.8 Tf [(N50= 223,917\) using CLCbio Genomic Workbench 8.0.2 \(Qiagen, Aarhus, Denmark\). Genes of mobile and prophage elements )] TJ ET BT 26.250 657.238 Td /F1 9.8 Tf [(in the reference )] TJ ET BT 96.704 657.238 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 144.927 657.238 Td /F1 9.8 Tf [( Poona M15X04725 genome were annotated using prokka v1.1 and PHAST tools and consequently )] TJ ET BT 26.250 645.333 Td /F1 9.8 Tf [(masked from mapping in CLCbio Genomic Workbench 8.0.2. SNPs were then called in coding and non-coding genome areas )] TJ ET BT 26.250 633.429 Td /F1 9.8 Tf [(using SAMtools mpileup \(v.1.2;)] TJ ET 0.267 0.267 0.267 rg BT 160.634 634.936 Td /F6 8.7 Tf [(10)] TJ ET 0.271 0.267 0.267 rg BT 170.272 633.429 Td /F1 9.8 Tf [(\) and converted into VCF matrix using bcftools \(v0.1.19; http://samtools.github.io/bcftools/\). )] TJ ET BT 26.250 621.524 Td /F1 9.8 Tf [(Variants were parsed using vcftools \(v.0.1.12b;)] TJ ET 0.267 0.267 0.267 rg BT 228.923 623.031 Td /F6 8.7 Tf [(11)] TJ ET 0.271 0.267 0.267 rg BT 238.561 621.524 Td /F1 9.8 Tf [(\) to include only high-quality SNPs \(hqSNPs\) with minimum position coverage )] TJ ET BT 26.250 609.619 Td /F1 9.8 Tf [(?30x and minimum quality of the base > 200 \(min-meanDP 30; minQ 200\), with InDels and the heterozygote calls excluded. )] TJ ET BT 26.250 597.714 Td /F1 9.8 Tf [(The Maximum Likelihood phylogenetic tree was generated based on hqSNPs calls under the Jukes-Cantor nucleotide )] TJ ET BT 26.250 585.810 Td /F1 9.8 Tf [(substitution model; with 100 bootstrap replicates.)] TJ ET BT 26.250 566.405 Td /F1 9.8 Tf [(At the US Centers for Disease Control and Prevention \(CDC\) LyveSET 1.1.4 SNP calling pipeline )] TJ ET BT 26.250 554.500 Td /F1 9.8 Tf [(\(http://www.github.com/lskatz/lyve-SET\) was applied with reads trimmed using fastx_trimmer 5 bases from 5 ends before )] TJ ET BT 26.250 542.595 Td /F1 9.8 Tf [(mapping single-end by SMALT. SNPs were called using Varscan at > 20x coverage, > 95% read support, and ?5 bp apart. The )] TJ ET BT 26.250 530.691 Td /F1 9.8 Tf [(draft assembly of FDA00009409 from the outbreak \(38 contigs\) was used as a reference without prophage masking.)] TJ ET BT 26.250 511.286 Td /F1 9.8 Tf [(Fourteen human and two cucumber )] TJ ET BT 182.864 511.286 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 231.088 511.286 Td /F1 9.8 Tf [( Poona isolates analyzed in this study exhibited 3 possibly related PFGE )] TJ ET BT 26.250 499.381 Td /F1 9.8 Tf [(patterns \(JL6X01.0018 [predominant], JL6X01.0375, JL6X01.0778\). The isolates were selected to include representatives of the )] TJ ET BT 26.250 487.476 Td /F1 9.8 Tf [(three above mentioned PFGE patterns and epidemiologically representative isolates with and without links to the cucumber )] TJ ET BT 26.250 475.572 Td /F1 9.8 Tf [(source. In the interest of the cost effectiveness, the WGS was limited to 16 isolates since the detected PFGE patterns were rare )] TJ ET BT 26.250 463.667 Td /F1 9.8 Tf [(in California and only few SNP differences were found among the sampled isolates. Of the 14 case-patients whose isolates )] TJ ET BT 26.250 451.762 Td /F1 9.8 Tf [(were analyzed, 10 \(71%\) reported consuming cucumbers during the week preceding their illness onsets, 2 case-patients denied )] TJ ET BT 26.250 439.857 Td /F1 9.8 Tf [(consuming any cucumbers, and cucumber exposure was unknown for 2 case-patients \(Table 1\). The cucumber exposures )] TJ ET BT 26.250 427.953 Td /F1 9.8 Tf [(reported by the 10 case-patients were all linked to Company A. One isolate \(M15X03586\) with a divergent PFGE pattern )] TJ ET BT 26.250 416.048 Td /F1 9.8 Tf [(\(JL6X01.0776\) served as an outlier in the phylogenetic analysis \(Table 1, Figure 1A\). Cucumber exposure for the patient was )] TJ ET BT 26.250 404.143 Td /F1 9.8 Tf [(unknown; the patient was not considered part of the outbreak cluster based on PFGE.)] TJ ET 0.965 0.965 0.965 rg 26.250 59.298 555.000 334.964 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 394.262 m 581.250 394.262 l 581.250 393.512 l 26.250 393.512 l f q 450.000 0 0 277.500 35.250 107.012 cm /I3 Do Q q 35.250 59.298 537.000 41.714 re W n Q Q q 450.000 0 0 277.500 35.250 107.012 cm /I3 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(2)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 156 0 obj << /Type /Annot /Subtype /Link /A 157 0 R /Border [0 0 0] /H /I /Rect [ 230.0153 736.8722 234.8339 745.6905 ] >> endobj 157 0 obj << /Type /Action >> endobj 158 0 obj << /Type /Annot /Subtype /Link /A 159 0 R /Border [0 0 0] /H /I /Rect [ 139.4865 724.9674 144.3052 733.7858 ] >> endobj 159 0 obj << /Type /Action >> endobj 160 0 obj << /Type /Annot /Subtype /Link /A 161 0 R /Border [0 0 0] /H /I /Rect [ 146.7145 724.9674 151.5332 733.7858 ] >> endobj 161 0 obj << /Type /Action >> endobj 162 0 obj << /Type /Annot /Subtype /Link /A 163 0 R /Border [0 0 0] /H /I /Rect [ 528.1269 724.9674 532.9456 733.7858 ] >> endobj 163 0 obj << /Type /Action >> endobj 164 0 obj << /Type /Annot /Subtype /Link /A 165 0 R /Border [0 0 0] /H /I /Rect [ 160.6343 634.1342 170.2716 642.9525 ] >> endobj 165 0 obj << /Type /Action >> endobj 166 0 obj << /Type /Annot /Subtype /Link /A 167 0 R /Border [0 0 0] /H /I /Rect [ 228.9232 622.2294 238.5606 631.0477 ] >> endobj 167 0 obj << /Type /Action >> endobj 168 0 obj << /Type /Annot /Subtype /Link /A 169 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 107.0123 485.2500 384.5123 ] >> endobj 169 0 obj << /Type /Action /S /URI /URI (http://currents.plos.org/outbreaks/files/2016/08/Fig1_large.jpg) >> endobj 170 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 370 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 45077>> stream JFIF``;CREATOR: gd-jpeg v1.0 (using IJG JPEG v80), quality = 82 C    !'"#%%%),($+!$%$C   $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$rX" }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?O|Eִ?iD<;l!0Ņ\1yKN;0z ּL#k:vs8u p*H/̽| eF4y447N,D)Qь'!uQU5nݾ|qJEs6λ&'Z/#A:TϬjeo:XGsp.C=mox˩ۤEaׯQߎֺ^@wF R?C@(U ~#iJ[R->nؓc^u *+}N;&<>TzEgjDϐDRrǠ8=BVR϶%\+4KEVlO2A@JET[XyxH @ tw~lk"k hBZF̰`F229EBr?_ϠzWS$E!pïQր/T.5hn BD8#^4^Tm#cjOP*j0c;K)S"jhoMp8hJSg8,B=qsRGwƲ. 8hBV2L BLڙCbbAL#O(J9u*TEyz}*ނ'Q締 <y(z*=/ނ&Q締 |뷚lPڤʤh"55++Ig9+r(/xڤN|<- >SpA3m zoޗA+/Hu(NǞ^Zy~Yk:bI-o$d `# `zAT]q|Qe>ik%͕wvm &ou]{Tun Idls/ʸU4QQۙ#pEC @=jJ((kƌʨ,p5F3h5'Q9#񲸏,'9qK#i r? 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Background coloring designates PFGE )] TJ ET BT 35.250 696.824 Td /F1 9.8 Tf [(pattern. Red boxes designate sequences generated by CDC; C- Maximum likelihood tree generated by Centers for Disease )] TJ ET BT 35.250 683.088 Td /F1 9.8 Tf [(Control and Prevention \(CDC\) for multistate investigation, including California. Red boxes designate selected isolates from )] TJ ET BT 35.250 669.352 Td /F1 9.8 Tf [(California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 331.831 555.000 251.809 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 583.640 m 581.250 583.640 l 581.250 582.890 l 26.250 582.890 l f 26.250 331.831 m 581.250 331.831 l 581.250 332.581 l 26.250 332.581 l f q 450.000 0 0 195.000 35.250 378.890 cm /I4 Do Q q 35.250 343.081 537.000 29.809 re W n 0.271 0.267 0.267 rg BT 35.250 363.366 Td /F6 9.8 Tf [(Fig. 2: Pairwise comparison of single nucleotide polymorphism \(SNP\) differences between Salmonella Poona )] TJ ET BT 35.250 351.462 Td /F6 9.8 Tf [(isolates from California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 -51.419 555.000 375.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 324.331 m 581.250 324.331 l 581.250 323.581 l 26.250 323.581 l f q 450.000 0 0 360.000 35.250 -45.419 cm /I6 Do Q q 35.250 -51.419 537.000 0.000 re W n Q Q q 15.000 -51.419 577.500 828.419 re W n 0.965 0.965 0.965 rg 26.250 591.140 555.000 185.860 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 591.140 m 581.250 591.140 l 581.250 591.890 l 26.250 591.890 l f q 35.250 602.390 537.000 174.610 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F6 9.8 Tf [(Fig. 1: Phylogeny of Salmonella enterica serovar Poona based on single nucleotide polymorphism \(SNP\) )] TJ ET BT 35.250 755.571 Td /F6 9.8 Tf [(differences discovered between the isolates by whole genome sequencing \(WGS\) vs. phylogeny based on PFGE )] TJ ET BT 35.250 743.667 Td /F6 9.8 Tf [(profiles clustering.)] TJ ET BT 35.250 724.297 Td /F1 9.8 Tf [(A- Pulsed-field gel electrophoresis \(PFGE\) dendrogram of the California isolates; B- Maximum likelihood tree generated by )] TJ ET BT 35.250 710.560 Td /F1 9.8 Tf [(California Department of Public Health \(CDPH\) for California outbreak isolates. Background coloring designates PFGE )] TJ ET BT 35.250 696.824 Td /F1 9.8 Tf [(pattern. Red boxes designate sequences generated by CDC; C- Maximum likelihood tree generated by Centers for Disease )] TJ ET BT 35.250 683.088 Td /F1 9.8 Tf [(Control and Prevention \(CDC\) for multistate investigation, including California. Red boxes designate selected isolates from )] TJ ET BT 35.250 669.352 Td /F1 9.8 Tf [(California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 331.831 555.000 251.809 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 583.640 m 581.250 583.640 l 581.250 582.890 l 26.250 582.890 l f 26.250 331.831 m 581.250 331.831 l 581.250 332.581 l 26.250 332.581 l f q 450.000 0 0 195.000 35.250 378.890 cm /I4 Do Q q 35.250 343.081 537.000 29.809 re W n 0.271 0.267 0.267 rg BT 35.250 363.366 Td /F6 9.8 Tf [(Fig. 2: Pairwise comparison of single nucleotide polymorphism \(SNP\) differences between Salmonella Poona )] TJ ET BT 35.250 351.462 Td /F6 9.8 Tf [(isolates from California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 -51.419 555.000 375.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 324.331 m 581.250 324.331 l 581.250 323.581 l 26.250 323.581 l f q 450.000 0 0 360.000 35.250 -45.419 cm /I8 Do Q q 35.250 -51.419 537.000 0.000 re W n Q Q q 15.000 -51.419 577.500 828.419 re W n 0.965 0.965 0.965 rg 26.250 591.140 555.000 185.860 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 591.140 m 581.250 591.140 l 581.250 591.890 l 26.250 591.890 l f q 35.250 602.390 537.000 174.610 re W n 0.271 0.267 0.267 rg BT 35.250 767.476 Td /F6 9.8 Tf [(Fig. 1: Phylogeny of Salmonella enterica serovar Poona based on single nucleotide polymorphism \(SNP\) )] TJ ET BT 35.250 755.571 Td /F6 9.8 Tf [(differences discovered between the isolates by whole genome sequencing \(WGS\) vs. phylogeny based on PFGE )] TJ ET BT 35.250 743.667 Td /F6 9.8 Tf [(profiles clustering.)] TJ ET BT 35.250 724.297 Td /F1 9.8 Tf [(A- Pulsed-field gel electrophoresis \(PFGE\) dendrogram of the California isolates; B- Maximum likelihood tree generated by )] TJ ET BT 35.250 710.560 Td /F1 9.8 Tf [(California Department of Public Health \(CDPH\) for California outbreak isolates. Background coloring designates PFGE )] TJ ET BT 35.250 696.824 Td /F1 9.8 Tf [(pattern. Red boxes designate sequences generated by CDC; C- Maximum likelihood tree generated by Centers for Disease )] TJ ET BT 35.250 683.088 Td /F1 9.8 Tf [(Control and Prevention \(CDC\) for multistate investigation, including California. Red boxes designate selected isolates from )] TJ ET BT 35.250 669.352 Td /F1 9.8 Tf [(California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 331.831 555.000 251.809 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 583.640 m 581.250 583.640 l 581.250 582.890 l 26.250 582.890 l f 26.250 331.831 m 581.250 331.831 l 581.250 332.581 l 26.250 332.581 l f q 450.000 0 0 195.000 35.250 378.890 cm /I4 Do Q q 35.250 343.081 537.000 29.809 re W n 0.271 0.267 0.267 rg BT 35.250 363.366 Td /F6 9.8 Tf [(Fig. 2: Pairwise comparison of single nucleotide polymorphism \(SNP\) differences between Salmonella Poona )] TJ ET BT 35.250 351.462 Td /F6 9.8 Tf [(isolates from California.)] TJ ET Q 0.965 0.965 0.965 rg 26.250 -51.419 555.000 375.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 324.331 m 581.250 324.331 l 581.250 323.581 l 26.250 323.581 l f q 450.000 0 0 360.000 35.250 -45.419 cm /I10 Do Q q 35.250 -51.419 537.000 0.000 re W n Q Q q 450.000 0 0 195.000 35.250 378.890 cm /I4 Do Q q 450.000 0 0 360.000 35.250 -45.419 cm /I12 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(3)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 201 0 obj << /Type /Annot /Subtype /Link /A 202 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 378.8902 485.2500 573.8902 ] >> endobj 202 0 obj << /Type /Action /S /URI /URI (http://currents.plos.org/outbreaks/files/2016/08/Fig2_large.jpg) >> endobj 203 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 260 /ColorSpace /DeviceRGB /Filter /DCTDecode /BitsPerComponent 8 /Length 43931>> stream JFIF``;CREATOR: gd-jpeg v1.0 (using IJG JPEG v80), quality = 82 C    !'"#%%%),($+!$%$C   $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$X" }!1AQa"q2#BR$3br %&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz w!1AQaq"2B #3Rbr $4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ?1F(Q(b +ˮo//=Ev\FԈ+*`Ln)lUz~m5_ w:zGjf1l폽`ͻUYu+d$K.}&F>{P^WSU1Nګ1(‡ ((&oMKcPTܒzK+8.M 5˒Pɑ(֨ou j\*7l#U6KPQ^>ukNk\ iOKæ}ny:_*=R:P;?eP{QNX#%o^Cm:IJ7pHb2=oMkP~VA{К +ۿ j/TvӍS}(V䂣}4W:_*^\ &"Q> +ȵ-R}.{u=a.U/e,il5 T]ST| rA @np#9ܾ{5~m5_v7ZIjal/cz(!5;>fmCXc }'(<@Zӵt50kpH捿 }^?^։$Zg3O *{КSUJ+gk~4p/ⴼۿ j/@Ex-کHeEAx5'I5NNk5YTIX W[jVo _Ug G)I^jK?m`?@Ey_yAMW*wVj2ił>u#97s@EyX쌍WUKU;kBMNյSKAx9/j +ȵ-JM ^EwK3ZJC1-~e{Y-4յU8Svm5_T[PՁi6mr.N{ ֚Ov5]P2 =Z5ծ`::eH?*~]i}joJʗd2q]j_RMKWV%w5ywm7(<2kWq["7e8`P^WSUͼ׮ B@X٨')$w@_JJ:62*Wj6V|~/ '?* +ȵ=NLӮoP%[xVD$([ګA .4kXys}.x_댮@=VP.!յE(ЛN)#5QEqGkkm=&3BosWRqf.ovO2nO@Wh~$[L' ci[# B3~$D\~uvm[×2G"tim`x 3]h.5V527W"V`Q5'$I?R״}ulb-ݨ]ɚhgu:}Fkul`Ҁ2uz%xj妌anJ?ySק@kпwgiz>o "C:=1עIXDdh'$E0Xx7Cʊ[i,UviZ[4]xkXɹ6ז܌v#4gWYa1̌ۂ=I?P<^հvV&h:ִu}W@.5'3זNw\?wxsӿg@U/ 7r?Z][0'[_r* etƶfbREb႑zVnn,ϖAoJzos᫉qGܫqSHd_\*$ 23^Ey'}ut+ڨLqǴпwkx@]hZJwS6sS=y\Ԯw³;\*othh&UccdBEI6ǃҀ< ]:찶%|8UnÚjhd^7ܫB<7c.|SJ%Lg Ag}k.^&p:n~CҪGWú>v,%p6xn1CјdӮq0`E lzrHO.B])3(w2[NW?yTPb/9Yh$4ܕogJ״uM2KXY3_!$ 6Sfxj]3RiI!lԕˎ=r>o W"x[r=*8+Ap1xZdid F|E_z *(ɼAkM&tc4(rx5|Wmt^I;|;F'Ǘ:ӽvqEP sSmKaY!}CW' rH/?c![Uw\A\CtzVg 0aPájt=*\)p>v߻#l ]_Phs^휊 2wn `*j_k ySʷE1dT ~8HӌJl- RN9$OXZ%ŕ) +"R01LP*x$Ȃ|2tۺq;s@Z5->Ԇ}9$`Fs#fM JL%,\O"(hC7y֭Q* g8dt:6uSOg 5Y$ꞧuOge"psSW.eQԅ}kIt?:S{-)!I|G1PlM6 ҭbH1HzO 8,%>N5ht҄o_v 3j} msϖWN '/RٙXo7o =&5S[g-?eG"57I̓qc3K`{` ?^үՄW;w1I0km=A`}n}k/Mg&O2ZsEnӕ'&?пi~p?Y?uеk&5^x:{º7nh3{8r0է@O+ }_LmK0V ^+IbP-EfRi'"zm?ج?[hMr&UфIH#=+Ibx@;A #V#34/ ho&K;o 8R?i~ew t(>sH0v1_zO d? 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WGS of cucumber isolates showed )] TJ ET BT 26.250 701.849 Td /F1 9.8 Tf [(close relatedness to the genomes of human isolates, with only 0-3 SNPs difference \(Figure 1B, 2\). The isolate with the )] TJ ET BT 26.250 689.944 Td /F1 9.8 Tf [(divergent PFGE pattern JL6X01.0776 was substantially different \(120-123 hqSNPs\) from the outbreak clade by WGS \(Figure )] TJ ET BT 26.250 678.039 Td /F1 9.8 Tf [(1A vs. 1B\). One outbreak isolate M15X04138 was sequenced and analyzed independently by CDC and CDPH laboratories; no )] TJ ET BT 26.250 666.135 Td /F1 9.8 Tf [(SNP differences in sequences were detected. Two additional isolates were sequenced by CDC, and the raw data was )] TJ ET BT 26.250 654.230 Td /F1 9.8 Tf [(processed through CDPH and CDC analysis pipelines. Both data analysis pipelines generated the same results \(Figure 1B vs. )] TJ ET BT 26.250 642.325 Td /F1 9.8 Tf [(1C\).)] TJ ET BT 26.250 622.920 Td /F1 9.8 Tf [(All outbreak isolates were assigned to the MLST sequence type 447 \(ST447\). The divergent outgroup strain had a single )] TJ ET BT 26.250 611.016 Td /F1 9.8 Tf [(mutation in )] TJ ET BT 76.648 611.016 Td /F5 9.8 Tf [(hemD)] TJ ET BT 102.651 611.016 Td /F1 9.8 Tf [( gene locus, which did not correspond to any known allele in the MLST database. There was only one entry of )] TJ ET BT 26.250 599.111 Td /F1 9.8 Tf [(ST447 in MLST database; )] TJ ET BT 142.226 599.111 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 193.160 599.111 Td /F1 9.8 Tf [(strain 3854/83 isolated in 1982 from a wild rodent in India.)] TJ ET BT 26.250 579.706 Td /F1 9.8 Tf [(The majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 337.850 579.706 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 386.074 579.706 Td /F1 9.8 Tf [( Pathogenicity Islands \(SPI\), with variability )] TJ ET BT 26.250 567.801 Td /F1 9.8 Tf [(mostly caused by the absence of SPI-2 or SPI-3, which encode Type III secretion system and invasion functions, respectively )] TJ ET BT 26.250 555.897 Td /F1 9.8 Tf [(\(Table 2\).)] TJ ET 0.965 0.965 0.965 rg 26.250 50.266 555.000 495.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 546.016 m 581.250 546.016 l 581.250 545.266 l 26.250 545.266 l f q 450.000 0 0 480.000 35.250 56.266 cm /I14 Do Q q 35.250 50.266 537.000 0.000 re W n Q Q q 15.000 50.266 577.500 726.734 re W n 0.965 0.965 0.965 rg 26.250 730.777 555.000 46.223 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 730.777 m 581.250 730.777 l 581.250 731.527 l 26.250 731.527 l f q 35.250 742.027 537.000 34.973 re W n 0.271 0.267 0.267 rg BT 35.250 766.011 Td /F6 9.8 Tf [(Table 1.)] TJ ET BT 71.559 766.011 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 157.174 766.011 Td /F1 9.8 Tf [( serotype Poona isolates, California \(N=17\): pulsed-field gel electrophoresis \(PFGE\) profiles, )] TJ ET BT 35.250 752.275 Td /F1 9.8 Tf [(whole genome sequencing \(WGS\)-based clustering, and patient-reported cucumber exposure.)] TJ ET Q BT 26.250 713.754 Td /F1 9.8 Tf [(All 16 isolates that were highly similar by PFGE also clustered together according to WGS. WGS of cucumber isolates showed )] TJ ET BT 26.250 701.849 Td /F1 9.8 Tf [(close relatedness to the genomes of human isolates, with only 0-3 SNPs difference \(Figure 1B, 2\). The isolate with the )] TJ ET BT 26.250 689.944 Td /F1 9.8 Tf [(divergent PFGE pattern JL6X01.0776 was substantially different \(120-123 hqSNPs\) from the outbreak clade by WGS \(Figure )] TJ ET BT 26.250 678.039 Td /F1 9.8 Tf [(1A vs. 1B\). One outbreak isolate M15X04138 was sequenced and analyzed independently by CDC and CDPH laboratories; no )] TJ ET BT 26.250 666.135 Td /F1 9.8 Tf [(SNP differences in sequences were detected. Two additional isolates were sequenced by CDC, and the raw data was )] TJ ET BT 26.250 654.230 Td /F1 9.8 Tf [(processed through CDPH and CDC analysis pipelines. Both data analysis pipelines generated the same results \(Figure 1B vs. )] TJ ET BT 26.250 642.325 Td /F1 9.8 Tf [(1C\).)] TJ ET BT 26.250 622.920 Td /F1 9.8 Tf [(All outbreak isolates were assigned to the MLST sequence type 447 \(ST447\). The divergent outgroup strain had a single )] TJ ET BT 26.250 611.016 Td /F1 9.8 Tf [(mutation in )] TJ ET BT 76.648 611.016 Td /F5 9.8 Tf [(hemD)] TJ ET BT 102.651 611.016 Td /F1 9.8 Tf [( gene locus, which did not correspond to any known allele in the MLST database. There was only one entry of )] TJ ET BT 26.250 599.111 Td /F1 9.8 Tf [(ST447 in MLST database; )] TJ ET BT 142.226 599.111 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 193.160 599.111 Td /F1 9.8 Tf [(strain 3854/83 isolated in 1982 from a wild rodent in India.)] TJ ET BT 26.250 579.706 Td /F1 9.8 Tf [(The majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 337.850 579.706 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 386.074 579.706 Td /F1 9.8 Tf [( Pathogenicity Islands \(SPI\), with variability )] TJ ET BT 26.250 567.801 Td /F1 9.8 Tf [(mostly caused by the absence of SPI-2 or SPI-3, which encode Type III secretion system and invasion functions, respectively )] TJ ET BT 26.250 555.897 Td /F1 9.8 Tf [(\(Table 2\).)] TJ ET 0.965 0.965 0.965 rg 26.250 50.266 555.000 495.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 546.016 m 581.250 546.016 l 581.250 545.266 l 26.250 545.266 l f q 450.000 0 0 480.000 35.250 56.266 cm /I16 Do Q q 35.250 50.266 537.000 0.000 re W n Q Q q 15.000 50.266 577.500 726.734 re W n 0.965 0.965 0.965 rg 26.250 730.777 555.000 46.223 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 730.777 m 581.250 730.777 l 581.250 731.527 l 26.250 731.527 l f q 35.250 742.027 537.000 34.973 re W n 0.271 0.267 0.267 rg BT 35.250 766.011 Td /F6 9.8 Tf [(Table 1.)] TJ ET BT 71.559 766.011 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 157.174 766.011 Td /F1 9.8 Tf [( serotype Poona isolates, California \(N=17\): pulsed-field gel electrophoresis \(PFGE\) profiles, )] TJ ET BT 35.250 752.275 Td /F1 9.8 Tf [(whole genome sequencing \(WGS\)-based clustering, and patient-reported cucumber exposure.)] TJ ET Q BT 26.250 713.754 Td /F1 9.8 Tf [(All 16 isolates that were highly similar by PFGE also clustered together according to WGS. WGS of cucumber isolates showed )] TJ ET BT 26.250 701.849 Td /F1 9.8 Tf [(close relatedness to the genomes of human isolates, with only 0-3 SNPs difference \(Figure 1B, 2\). The isolate with the )] TJ ET BT 26.250 689.944 Td /F1 9.8 Tf [(divergent PFGE pattern JL6X01.0776 was substantially different \(120-123 hqSNPs\) from the outbreak clade by WGS \(Figure )] TJ ET BT 26.250 678.039 Td /F1 9.8 Tf [(1A vs. 1B\). One outbreak isolate M15X04138 was sequenced and analyzed independently by CDC and CDPH laboratories; no )] TJ ET BT 26.250 666.135 Td /F1 9.8 Tf [(SNP differences in sequences were detected. Two additional isolates were sequenced by CDC, and the raw data was )] TJ ET BT 26.250 654.230 Td /F1 9.8 Tf [(processed through CDPH and CDC analysis pipelines. Both data analysis pipelines generated the same results \(Figure 1B vs. )] TJ ET BT 26.250 642.325 Td /F1 9.8 Tf [(1C\).)] TJ ET BT 26.250 622.920 Td /F1 9.8 Tf [(All outbreak isolates were assigned to the MLST sequence type 447 \(ST447\). The divergent outgroup strain had a single )] TJ ET BT 26.250 611.016 Td /F1 9.8 Tf [(mutation in )] TJ ET BT 76.648 611.016 Td /F5 9.8 Tf [(hemD)] TJ ET BT 102.651 611.016 Td /F1 9.8 Tf [( gene locus, which did not correspond to any known allele in the MLST database. There was only one entry of )] TJ ET BT 26.250 599.111 Td /F1 9.8 Tf [(ST447 in MLST database; )] TJ ET BT 142.226 599.111 Td /F5 9.8 Tf [(Salmonella )] TJ ET BT 193.160 599.111 Td /F1 9.8 Tf [(strain 3854/83 isolated in 1982 from a wild rodent in India.)] TJ ET BT 26.250 579.706 Td /F1 9.8 Tf [(The majority of the outbreak-related isolates possessed the same set of )] TJ ET BT 337.850 579.706 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 386.074 579.706 Td /F1 9.8 Tf [( Pathogenicity Islands \(SPI\), with variability )] TJ ET BT 26.250 567.801 Td /F1 9.8 Tf [(mostly caused by the absence of SPI-2 or SPI-3, which encode Type III secretion system and invasion functions, respectively )] TJ ET BT 26.250 555.897 Td /F1 9.8 Tf [(\(Table 2\).)] TJ ET 0.965 0.965 0.965 rg 26.250 50.266 555.000 495.750 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 546.016 m 581.250 546.016 l 581.250 545.266 l 26.250 545.266 l f q 450.000 0 0 480.000 35.250 56.266 cm /I18 Do Q q 35.250 50.266 537.000 0.000 re W n Q Q q 450.000 0 0 480.000 35.250 56.266 cm /I20 Do Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(4)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 224 0 obj << /Type /Annot /Subtype /Link /A 225 0 R /Border [0 0 0] /H /I /Rect [ 35.2500 56.2657 485.2500 536.2657 ] >> endobj 225 0 obj << /Type /Action /S /URI /URI (http://currents.plos.org/outbreaks/files/2016/08/Table2-1.png) >> endobj 226 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 640 /Filter /FlateDecode /DecodeParms << /Predictor 15 /Colors 1 /Columns 600 /BitsPerComponent 8>> /ColorSpace /DeviceGray /BitsPerComponent 8 /Length 1628>> stream xA 0 =" ʝ<2;EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$EX$ۚ1 endstream endobj 227 0 obj << /Type /XObject /Subtype /Image /Width 600 /Height 640 /SMask 226 0 R /Filter /FlateDecode /DecodeParms << /Predictor 15 /Colors 3 /Columns 600 /BitsPerComponent 8>> /ColorSpace /DeviceRGB /BitsPerComponent 8 /Length 104898>> stream xy@o Kfid5XYieӘmSX2}lՏf{Ffdf"n"?>.e}s8<,{y@ @ D@@ a @@ a @@ a @@ a @@ a HIځǏ%8::K "#$TTTJ "A4#DDD\R^@ @KK@ _B dX!@50B dX!@50B dX3\a||CCCӧOTVVΛ7ҥK\.̙3/_677ojj)c/_tpprȹsN:o޼3g,Zʊ$$$P(1c,Z(55511ť1##رcŋ˗ؼxb߾}ϟ^sWRRc [MMM&I$1LRRϟOlٲL'''##$Ɣx&//n:ee`EEŮ.;wVVVFFF455ݻw4''޽YYYP__NP֭[7vXdzɒ%L&ꅅSN;wnDDٳuttzϟۏ3?R^^^UUeMMϟ?[[[GFF[Ғ^~?ckhhs玌-2X__iӦB@]]ݽ{ '##vuuXYYh42y}w| I]]]DDƍ*++dëMaƜkhh7hDDD u;v@fgg[YY544HKKٳ͞={ƍ<o/>ѣ\\\Μ9#--B–/_hlllmm]pann_|)..deeXXXTTTؘCvFFF...뱱h4N+))رN;/++`0eee[[[oܸ1k,3mڴ{Z[[544TTTΝ <bʕ<2**gԩ;;;;;֊ ))Cxg j7Y[[߾};777** y׮];vlGGٳ׮]]...ӧOGF\)ݺu+$$DUUb'O~2bll\PP`gg7sV@ P~-)))5j`ܸqnxox[nxs=z<o߾} ۷D>] .骪111_NNNt皓d"|@(Z[[=zm۶UVNGGǦ GGǎwVXQ__?jԨs2S޼y311_-++p8^RQQ155p8SNUSS;tرc߾]vpiӦ>}ikkWWW#8A FkkklllTT޵k $''BF)''譭544Y,֍7444߾}[QQQTT,ƒ丸;%%NWUU!,--{=w\FF`0EEE]]]''[n=9|˗/dr}}=ǫ#<|pʔ)(Аo*88x۶m!//>}f͚%Kdgg#,rJzzbO z fɅ7o|̙3gϞQTٳG2uTyyygX^^ #,, ??_YYyʕo߾={\ 7nlEEp))Oegg;::޹sGAA(j޼ymmm;w̔KHHbNNNaaa 3gpκ͍bݺu éGEE-\1LXX#U<$5F,{U{{{Q^f-s鴴Й3g2L˗yŋ "0L6-8rȑݻwGFFnݺCRSS33roݺ^xQДJEn_xǏSN0᫹T*B" ?NHHN2ĉSNmllj $H q{{ׯ####V\)bڽC:?_rQmhh,,,\v͛q8ܗ/_&Lեd"8r%K.Z,''eʔ)jjjK.555---R666.))`07of0555K.0aZ\\\aahϟmmm'NX\\<}3fttttttL:UZZzJJJ6-##e``,a0755M4iɪH̖4iVWW722>}V\\\MMMppږ-[޽{˖-366.**iiiinn?~* "Hƍ3g΢E SSS.\bqggg# 222f͚0al*))ihhL<۷999;w믿8ĉg͚UYYikkԴ` uuI&!iVVVx<4iҴi&LP__ٹ{n~򭞞^PPЪUeddjkk߼ykOOO܎===.F'M###aÆ:::N:c! =AL&311133sٮrrr$ BM6F9::jjjN4~̘1Ǐz SRRN8_\\|G/_N233f͚]TT4jԨm۶<'N\RRޞcll|%/ȿՆ vG<==eddh4իWxFF,//GG= r\GGǛ7ojiicH ؆iٿHHH_Yw_HII!X,̙3k;v?~|VZecco8!!!?" 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VTT +V244}ooҥKߩJzzzFFFF,3 7H͛7IbM6a}./// ܹsXK)))"N+**2 )q?]veeeI{+h")77l؀u񲳳 >H\.Wz~Q)t nCAЧB hLcW[[{sA2 яDjjjQQQllm]&##cǎ8l`X`ѱ֊RQQ222 \.Bddd(jiij*sGGGl<ۚ5k+bbb޿/^| A^GPQQqg ,+""BWWW__F) L GQĄBR(===@䔐Nt2U޽{gΜW_UTT#RWW7""b$[B4p8 Çܹ#ʺܹcW\Sb[Ɇj:NRZZZ:+˫/^R`uvSSD檪;wJJJ֢(Z\\Lb_€eٳ+Wb|>J UHtȑ?,p8ث DGD]]]b*fRR_|#AUURF%ׯ_^ supfTjss3WA0ݮN޹s쫶L&M h C``w[[۱cTUUL˗/׮]d2WX! 322f͚bBCC ԉttt8::*)) B"}o}}.~9DGGGZZ>@QQqŊS|Fͺ˗/HӧO Ǐ:u3::9..bXċ2###ϟvZߦ`@;𸱱1$$6**Þ̈@A@{f^^^R掣(~z=D>///_ ⒀JNN?ƍsrrwyPx]"x}#G8pl6;>>,..M h YYYYY+V+**^v}}}555zzzZZZ۷o>yd___nn.D9RRRYϟo``P^^F***6l ^''''''u>SRR8_{n0b,,,z{{333@_A455.]e˖3f̟?SKKHGH"jjjQRWW ;y$؀H$蘘/ .: HaaӔ)S@$@*,\5RT b_~o={6 O>mnng6FFFSNݷo߆ tuuwy!+WLIIڲe #_zgϞAl?RwwV"HP|}}߿?t/_:;;1d29""BNN ,AOO䜜 .D"999vuu?~ r\t v3gzzzX#e6vǺ_ŋ/_ nJhVVVvvv`` ,|e`````19997ol6BP(>TWW^UR;99xRFGGJ6@ P(***N277ӫ~k׆noiiyZ,ÿ3gmܸ155D<{oݺ-#3g>>"޽{{SԨBA&N'322<*~]vxD) kkǏ3 |pF'~l6;,,l޼yWϟP( lt]]Ś8qbmm[[['O|++\h6mrrr} [A GH$Gl ޔ?GqT뫯 F!>ollIz+Ě{&)x:o $L!>}# .\+A룁cAИ6z\r%]ߟrW\9mڴhuuuh+HF_~qFEDD.]bٻvbիEEEK,quu}&M233>}AkYYYfrb>?o<VPxu77q\(=ztǎ&&&ؓUUUEEE'O633#Ht:A[YYq8b@pBmmm\.755AwwVi4Ç+++On``JbgA---[[7M9 nnnѺx<ޣG^~ ɰKKK!s!m۶ԓTUU-Y䭇 A#,))ٴid֭"L&K,0CCCeee+Wh>>>&&&8Cs9}X,>U'N(++ [ʐ[P(AH$fjjiӦUUڤ$VP,gggw ڕ 46PTTt ///SSSP'^tICC4## `"9sP544pݻwAh+i4aA|=@EEfGEEYYY%$$\ -ɓ\.q_MMMMMMGXO yyy _skmmmz>Fkkk|h^ @HI&uvvرcĉ7; " |ݾ}ԩSkK$UUUx<ǫŇnt#_VVJ۾}W@lOCKoii|ŋRr|@͠*))ϯh(p܈P(G&KKKsssLfII 9vZlb t钃CXXY,V@@@ooƍjkk?^TThRRRUUUJJJPPPOOO@@'@iɶBƍ999999 PSS(F)..rvvvuuu?9sftt4H%effj;;;s$qgo$y|w;vHu177///7nÇ糲x<^tt4~nP}vZb^ãG qRR WA___[[{V🜸3g;7`2//oܸq:::˖-444\LQQQTTTqq TTTO ;ՓD"01ZyC~B 522z6d2B۷oxb|̗2pwnԩ  h( b%b{A˗ e׮]X<r-,,9fff~~~CM5Gֶt$@ee ٓk1#$@пgkjj|g$@ ȼ)D:sP(711E"ׯAΛ7 VBwwEutt@ବ,)en]=x𠥥AsR(7$>|M 66(&Jcr0~[+s)P}}"//|Cqe˖uttH9EC/ XYz͉Db@@@III``۪}}}O?D $!aEܖOx<l2A:t N> R[[f322 X,|իWt:t:BYYYXXX,\D"YYY&%%:99%$$Ā/@N>⒝f777A -Zdnn/ Fsssss3ŢSNp8t:|߭_t:&:::((hpp0777$$NTVV\A6Uiiy@喗Gi4@ /###[ZZAbbIlv[[(AEE{ѢE/_h?d2W(VVVr8Ǐéd2(bFgggyy#Gtuu`K#ccccrJ6mjj K\Abbbbbb?A¦ǏT\AAA|R5:t:@ؿDڿ?X+ KKK}||݇HV\ :D'08z"RYY<GVWWbۙLs˗/9 "<DMMMK---/^|)888xbWE"QCCC?o{{;xގ=~V)7+dppիWwNmooǎNBkkkWWׯVBAQ+dϭU~-%%e$5/((hjjjjjjhhaҍBMW}W[p3YouT#Ie'''ddd;NHΉЕW@:=v2/222!!a_%ѩ KE,)5OIIpppihhaoz sEDD@4B+Ĵi>uE:\ttt\~D"i|:::;ozM 9s| 466/ZHVa=?!A1eʔO}b57oU >)S|L>! 3_>AA `CAi! 4BAcl!1 6AИB hL !A4 `CAi! 4BAcl!1 6AИB hL !A4 `CAi! 4BAcl!1 6AИB hL !A4 `CAi! 4BAcl!1 6AИB hL !A4 `CAi! 4BAc٣^ endstream endobj 238 0 obj << /Type /Page /Parent 3 0 R /Annots [ 240 0 R 242 0 R 244 0 R 246 0 R 248 0 R 250 0 R 252 0 R 254 0 R 256 0 R 258 0 R 260 0 R 262 0 R 264 0 R 266 0 R 268 0 R 270 0 R 272 0 R 274 0 R 276 0 R 278 0 R 280 0 R ] /Contents 239 0 R >> endobj 239 0 obj << /Length 28120 >> stream q 15.000 22.564 577.500 754.436 re W n 0.965 0.965 0.965 rg 26.250 744.514 555.000 32.486 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 744.514 m 581.250 744.514 l 581.250 745.264 l 26.250 745.264 l f q 35.250 755.764 537.000 21.236 re W n 0.271 0.267 0.267 rg BT 35.250 766.011 Td /F6 9.8 Tf [(Table 2.)] TJ ET BT 71.559 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 119.782 766.011 Td /F1 9.8 Tf [( Pathogenicity Islands in outbreak-related )] TJ ET BT 301.327 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 349.551 766.011 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET Q BT 26.250 727.490 Td /F1 9.8 Tf [(All isolates possessed phage elements homologous to )] TJ ET BT 263.614 727.490 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 311.837 727.490 Td /F1 9.8 Tf [( phage Fels-1 \(NC_010391\) and partial )] TJ ET BT 483.623 727.490 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 547.027 727.490 Td /F1 9.8 Tf [( phage )] TJ ET BT 26.250 715.585 Td /F1 9.8 Tf [(P4 \(NC_001609\). Four isolates harbored the genes homologous to an additional bacteriophage of either of the following types: )] TJ ET BT 26.250 703.680 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 94.529 703.680 Td /F1 9.8 Tf [( phage S-SKS1 \(NC_020851\), )] TJ ET BT 228.387 703.680 Td /F5 9.8 Tf [(Ectocarpus )] TJ ET BT 279.867 703.680 Td /F5 9.8 Tf [(siliculosus)] TJ ET BT 324.288 703.680 Td /F1 9.8 Tf [( virus 1 \(NC_002687\), )] TJ ET BT 422.363 703.680 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 485.767 703.680 Td /F1 9.8 Tf [( phage ES18 )] TJ ET BT 26.250 691.776 Td /F1 9.8 Tf [(\(NC_006949\), or )] TJ ET BT 101.569 691.776 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 164.973 691.776 Td /F1 9.8 Tf [( phage P1 \(NC_005856\). Interestingly, phage genes found in one of the isolates were )] TJ ET BT 26.250 679.871 Td /F1 9.8 Tf [(homologous to S-SKS1 bacteriophage, which was described as a virus infecting )] TJ ET BT 373.613 679.871 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 441.892 679.871 Td /F1 9.8 Tf [(, a genus of Cyanobacteria )] TJ ET BT 26.250 667.966 Td /F1 9.8 Tf [(known to be one of the predominant prokaryotic components of the oceans picophytoplankton)] TJ ET 0.267 0.267 0.267 rg BT 432.718 669.473 Td /F6 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 442.355 667.966 Td /F1 9.8 Tf [(. Another isolate possessed )] TJ ET BT 26.250 656.061 Td /F1 9.8 Tf [(genes homologous to EsV-1 bacteriophage, characterized previously as a bacteriophage of the marine filamentous brown algae )] TJ ET BT 26.250 644.157 Td /F5 9.8 Tf [(Ectocarpus siliculosus)] TJ ET 0.267 0.267 0.267 rg BT 122.151 645.664 Td /F6 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 131.788 644.157 Td /F1 9.8 Tf [(. The sequences identified as incomplete S-SKS1 or EsV-1 phages were integrated into the larger )] TJ ET BT 26.250 632.252 Td /F1 9.8 Tf [(contigs with clear )] TJ ET BT 103.733 632.252 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 151.957 632.252 Td /F1 9.8 Tf [( identity according to BLAST search. The marine phage-like sequences were also common in )] TJ ET BT 26.250 620.347 Td /F1 9.8 Tf [(other )] TJ ET BT 51.181 620.347 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 136.796 620.347 Td /F1 9.8 Tf [( serotypes found in NCBI database and seem to represent a part of the )] TJ ET BT 445.714 620.347 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 493.938 620.347 Td /F1 9.8 Tf [( accessory genome.)] TJ ET BT 26.250 600.942 Td /F1 9.8 Tf [(WGS has proven to be a powerful tool for the investigation of outbreaks caused by )] TJ ET BT 384.475 600.942 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 470.089 600.942 Td /F1 9.8 Tf [(, providing great )] TJ ET BT 26.250 589.038 Td /F1 9.8 Tf [(epidemiological concordance and higher resolution than PFGE, the traditional method for )] TJ ET BT 412.642 589.038 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 460.866 589.038 Td /F1 9.8 Tf [( strain subtyping)] TJ ET 0.267 0.267 0.267 rg BT 531.856 590.545 Td /F6 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 541.493 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 543.902 590.545 Td /F6 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 553.540 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 555.949 590.545 Td /F6 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 565.586 589.038 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 577.133 Td /F1 9.8 Tf [(In our study, WGS confirmed the genetic relatedness of strains with similar PFGE patterns. Outbreak-related isolates exhibited )] TJ ET BT 26.250 565.228 Td /F1 9.8 Tf [(3 PFGE patterns which complicated assignment of the isolates to a single outbreak source. Particularly, isolate M15X04477 )] TJ ET BT 26.250 553.323 Td /F1 9.8 Tf [(with the pattern JL6X01.0778 differed by 3 bands from the predominant outbreak PFGE-type, thus introducing a higher degree )] TJ ET BT 26.250 541.419 Td /F1 9.8 Tf [(of uncertainty regarding the inclusion of this isolate in the outbreak cluster even though the epidemiologic information supported )] TJ ET BT 26.250 529.514 Td /F1 9.8 Tf [(a possible link to the outbreak. WGS, on the other hand, showed unambiguous clustering of all epidemiologically-related )] TJ ET BT 26.250 517.609 Td /F1 9.8 Tf [(isolates together and clarified genetic relatedness of M15X04477 to the other outbreak isolates. Two case-patients denying )] TJ ET BT 26.250 505.704 Td /F1 9.8 Tf [(cucumber consumption also clustered with the outbreak, which could be explained by possible secondary transmission, cross-)] TJ ET BT 26.250 493.800 Td /F1 9.8 Tf [(contamination of other foods, poor food history recall, or case-patients not noticing cucumbers as part of a multi-ingredient meal )] TJ ET BT 26.250 481.895 Td /F1 9.8 Tf [(such as a salad or a sandwich. In support of the high reproducibility of the method, the results of WGS performed by two )] TJ ET BT 26.250 469.990 Td /F1 9.8 Tf [(different laboratories were identical. Additionally, WGS provided information about the MLST sequence type and acquired )] TJ ET BT 26.250 458.085 Td /F1 9.8 Tf [(components of the genome, allowing for more comprehensive characterization of the isolates. Though MLST usually provides )] TJ ET BT 26.250 446.181 Td /F1 9.8 Tf [(poor resolution between )] TJ ET BT 133.012 446.181 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 181.236 446.181 Td /F1 9.8 Tf [( strains, it is helpful for understanding the global epidemiology of infectious clones)] TJ ET 0.267 0.267 0.267 rg BT 533.484 447.688 Td /F6 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 543.121 446.181 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 434.276 Td /F1 9.8 Tf [(Diversity in accessory genes of the studied isolates demonstrates the plasticity of the )] TJ ET BT 394.741 434.276 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 442.965 434.276 Td /F1 9.8 Tf [( Poona genome which is )] TJ ET BT 26.250 422.371 Td /F1 9.8 Tf [(potentially responsible for variations in PFGE patterns that do not necessarily reflect the phylogenetic distance between the )] TJ ET BT 26.250 410.466 Td /F1 9.8 Tf [(isolates)] TJ ET 0.267 0.267 0.267 rg BT 59.303 411.974 Td /F6 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 68.940 410.466 Td /F1 9.8 Tf [(. Since the loss/acquisition of mobile elements does not affect genome-wide SNPs calling, it explains higher )] TJ ET BT 26.250 398.562 Td /F1 9.8 Tf [(evolutionary congruence of WGS-inferred phylogeny compared with PFGE. Noteworthy, WGS results were acquired in 7 )] TJ ET BT 26.250 386.657 Td /F1 9.8 Tf [(working days from the time of pure culture reception by the laboratory. Our experience suggests that WGS can be used for )] TJ ET BT 26.250 374.752 Td /F1 9.8 Tf [(routine epidemiological subtyping.)] TJ ET BT 26.250 338.150 Td /F6 12.0 Tf [(Conclusions)] TJ ET BT 26.250 318.195 Td /F1 9.8 Tf [(Real-time PFGE and WGS genotyping results from this )] TJ ET BT 266.831 318.195 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 315.055 318.195 Td /F1 9.8 Tf [( Poona outbreak provided concordant assignment of isolates )] TJ ET BT 26.250 306.291 Td /F1 9.8 Tf [(to the outbreak cluster by both methods. However, the WGS allowed for a more unequivocal determination of the genetic )] TJ ET BT 26.250 294.386 Td /F1 9.8 Tf [(relatedness of the isolates than PFGE. WGS provided additional information regarding MLST type, pathogenicity genes, and )] TJ ET BT 26.250 282.481 Td /F1 9.8 Tf [(bacteriophage content of the isolates. WGS data obtained independently at two laboratories showed complete agreement.)] TJ ET BT 26.250 245.879 Td /F6 12.0 Tf [(Data Availability Statement)] TJ ET BT 26.250 225.924 Td /F1 9.8 Tf [(The raw reads for sequenced here samples were submitted to NCBI SRA archive under accession numbers SRR2566826- )] TJ ET BT 26.250 214.020 Td /F1 9.8 Tf [(SRR2566828, SRR2566830, SRR3086799- SRR3086806, SRR3086823, SRR3086828, SRR3086831, SRR3086834, )] TJ ET BT 26.250 202.115 Td /F1 9.8 Tf [(SRR3095255 \(see Table 1 for details\).)] TJ ET BT 26.250 165.512 Td /F6 12.0 Tf [(Competing Interest Statement)] TJ ET BT 26.250 145.558 Td /F1 9.8 Tf [(The authors have declared that no competing interests exist.)] TJ ET BT 26.250 108.956 Td /F6 12.0 Tf [(Corresponding Authors)] TJ ET BT 26.250 89.001 Td /F1 9.8 Tf [(Vishnu Chaturvedi: Vishnu.Chaturvedi@cdph.ca.gov)] TJ ET BT 26.250 69.597 Td /F1 9.8 Tf [(Varvara K. Kozyreva: Varvara.Kozyreva@cdph.ca.gov)] TJ ET Q q 15.000 22.564 577.500 754.436 re W n 0.965 0.965 0.965 rg 26.250 744.514 555.000 32.486 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 744.514 m 581.250 744.514 l 581.250 745.264 l 26.250 745.264 l f q 35.250 755.764 537.000 21.236 re W n 0.271 0.267 0.267 rg BT 35.250 766.011 Td /F6 9.8 Tf [(Table 2.)] TJ ET BT 71.559 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 119.782 766.011 Td /F1 9.8 Tf [( Pathogenicity Islands in outbreak-related )] TJ ET BT 301.327 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 349.551 766.011 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET Q BT 26.250 727.490 Td /F1 9.8 Tf [(All isolates possessed phage elements homologous to )] TJ ET BT 263.614 727.490 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 311.837 727.490 Td /F1 9.8 Tf [( phage Fels-1 \(NC_010391\) and partial )] TJ ET BT 483.623 727.490 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 547.027 727.490 Td /F1 9.8 Tf [( phage )] TJ ET BT 26.250 715.585 Td /F1 9.8 Tf [(P4 \(NC_001609\). Four isolates harbored the genes homologous to an additional bacteriophage of either of the following types: )] TJ ET BT 26.250 703.680 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 94.529 703.680 Td /F1 9.8 Tf [( phage S-SKS1 \(NC_020851\), )] TJ ET BT 228.387 703.680 Td /F5 9.8 Tf [(Ectocarpus )] TJ ET BT 279.867 703.680 Td /F5 9.8 Tf [(siliculosus)] TJ ET BT 324.288 703.680 Td /F1 9.8 Tf [( virus 1 \(NC_002687\), )] TJ ET BT 422.363 703.680 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 485.767 703.680 Td /F1 9.8 Tf [( phage ES18 )] TJ ET BT 26.250 691.776 Td /F1 9.8 Tf [(\(NC_006949\), or )] TJ ET BT 101.569 691.776 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 164.973 691.776 Td /F1 9.8 Tf [( phage P1 \(NC_005856\). Interestingly, phage genes found in one of the isolates were )] TJ ET BT 26.250 679.871 Td /F1 9.8 Tf [(homologous to S-SKS1 bacteriophage, which was described as a virus infecting )] TJ ET BT 373.613 679.871 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 441.892 679.871 Td /F1 9.8 Tf [(, a genus of Cyanobacteria )] TJ ET BT 26.250 667.966 Td /F1 9.8 Tf [(known to be one of the predominant prokaryotic components of the oceans picophytoplankton)] TJ ET 0.267 0.267 0.267 rg BT 432.718 669.473 Td /F6 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 442.355 667.966 Td /F1 9.8 Tf [(. Another isolate possessed )] TJ ET BT 26.250 656.061 Td /F1 9.8 Tf [(genes homologous to EsV-1 bacteriophage, characterized previously as a bacteriophage of the marine filamentous brown algae )] TJ ET BT 26.250 644.157 Td /F5 9.8 Tf [(Ectocarpus siliculosus)] TJ ET 0.267 0.267 0.267 rg BT 122.151 645.664 Td /F6 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 131.788 644.157 Td /F1 9.8 Tf [(. The sequences identified as incomplete S-SKS1 or EsV-1 phages were integrated into the larger )] TJ ET BT 26.250 632.252 Td /F1 9.8 Tf [(contigs with clear )] TJ ET BT 103.733 632.252 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 151.957 632.252 Td /F1 9.8 Tf [( identity according to BLAST search. The marine phage-like sequences were also common in )] TJ ET BT 26.250 620.347 Td /F1 9.8 Tf [(other )] TJ ET BT 51.181 620.347 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 136.796 620.347 Td /F1 9.8 Tf [( serotypes found in NCBI database and seem to represent a part of the )] TJ ET BT 445.714 620.347 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 493.938 620.347 Td /F1 9.8 Tf [( accessory genome.)] TJ ET BT 26.250 600.942 Td /F1 9.8 Tf [(WGS has proven to be a powerful tool for the investigation of outbreaks caused by )] TJ ET BT 384.475 600.942 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 470.089 600.942 Td /F1 9.8 Tf [(, providing great )] TJ ET BT 26.250 589.038 Td /F1 9.8 Tf [(epidemiological concordance and higher resolution than PFGE, the traditional method for )] TJ ET BT 412.642 589.038 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 460.866 589.038 Td /F1 9.8 Tf [( strain subtyping)] TJ ET 0.267 0.267 0.267 rg BT 531.856 590.545 Td /F6 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 541.493 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 543.902 590.545 Td /F6 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 553.540 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 555.949 590.545 Td /F6 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 565.586 589.038 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 577.133 Td /F1 9.8 Tf [(In our study, WGS confirmed the genetic relatedness of strains with similar PFGE patterns. Outbreak-related isolates exhibited )] TJ ET BT 26.250 565.228 Td /F1 9.8 Tf [(3 PFGE patterns which complicated assignment of the isolates to a single outbreak source. Particularly, isolate M15X04477 )] TJ ET BT 26.250 553.323 Td /F1 9.8 Tf [(with the pattern JL6X01.0778 differed by 3 bands from the predominant outbreak PFGE-type, thus introducing a higher degree )] TJ ET BT 26.250 541.419 Td /F1 9.8 Tf [(of uncertainty regarding the inclusion of this isolate in the outbreak cluster even though the epidemiologic information supported )] TJ ET BT 26.250 529.514 Td /F1 9.8 Tf [(a possible link to the outbreak. WGS, on the other hand, showed unambiguous clustering of all epidemiologically-related )] TJ ET BT 26.250 517.609 Td /F1 9.8 Tf [(isolates together and clarified genetic relatedness of M15X04477 to the other outbreak isolates. Two case-patients denying )] TJ ET BT 26.250 505.704 Td /F1 9.8 Tf [(cucumber consumption also clustered with the outbreak, which could be explained by possible secondary transmission, cross-)] TJ ET BT 26.250 493.800 Td /F1 9.8 Tf [(contamination of other foods, poor food history recall, or case-patients not noticing cucumbers as part of a multi-ingredient meal )] TJ ET BT 26.250 481.895 Td /F1 9.8 Tf [(such as a salad or a sandwich. In support of the high reproducibility of the method, the results of WGS performed by two )] TJ ET BT 26.250 469.990 Td /F1 9.8 Tf [(different laboratories were identical. Additionally, WGS provided information about the MLST sequence type and acquired )] TJ ET BT 26.250 458.085 Td /F1 9.8 Tf [(components of the genome, allowing for more comprehensive characterization of the isolates. Though MLST usually provides )] TJ ET BT 26.250 446.181 Td /F1 9.8 Tf [(poor resolution between )] TJ ET BT 133.012 446.181 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 181.236 446.181 Td /F1 9.8 Tf [( strains, it is helpful for understanding the global epidemiology of infectious clones)] TJ ET 0.267 0.267 0.267 rg BT 533.484 447.688 Td /F6 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 543.121 446.181 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 434.276 Td /F1 9.8 Tf [(Diversity in accessory genes of the studied isolates demonstrates the plasticity of the )] TJ ET BT 394.741 434.276 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 442.965 434.276 Td /F1 9.8 Tf [( Poona genome which is )] TJ ET BT 26.250 422.371 Td /F1 9.8 Tf [(potentially responsible for variations in PFGE patterns that do not necessarily reflect the phylogenetic distance between the )] TJ ET BT 26.250 410.466 Td /F1 9.8 Tf [(isolates)] TJ ET 0.267 0.267 0.267 rg BT 59.303 411.974 Td /F6 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 68.940 410.466 Td /F1 9.8 Tf [(. Since the loss/acquisition of mobile elements does not affect genome-wide SNPs calling, it explains higher )] TJ ET BT 26.250 398.562 Td /F1 9.8 Tf [(evolutionary congruence of WGS-inferred phylogeny compared with PFGE. Noteworthy, WGS results were acquired in 7 )] TJ ET BT 26.250 386.657 Td /F1 9.8 Tf [(working days from the time of pure culture reception by the laboratory. Our experience suggests that WGS can be used for )] TJ ET BT 26.250 374.752 Td /F1 9.8 Tf [(routine epidemiological subtyping.)] TJ ET BT 26.250 338.150 Td /F6 12.0 Tf [(Conclusions)] TJ ET BT 26.250 318.195 Td /F1 9.8 Tf [(Real-time PFGE and WGS genotyping results from this )] TJ ET BT 266.831 318.195 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 315.055 318.195 Td /F1 9.8 Tf [( Poona outbreak provided concordant assignment of isolates )] TJ ET BT 26.250 306.291 Td /F1 9.8 Tf [(to the outbreak cluster by both methods. However, the WGS allowed for a more unequivocal determination of the genetic )] TJ ET BT 26.250 294.386 Td /F1 9.8 Tf [(relatedness of the isolates than PFGE. WGS provided additional information regarding MLST type, pathogenicity genes, and )] TJ ET BT 26.250 282.481 Td /F1 9.8 Tf [(bacteriophage content of the isolates. WGS data obtained independently at two laboratories showed complete agreement.)] TJ ET BT 26.250 245.879 Td /F6 12.0 Tf [(Data Availability Statement)] TJ ET BT 26.250 225.924 Td /F1 9.8 Tf [(The raw reads for sequenced here samples were submitted to NCBI SRA archive under accession numbers SRR2566826- )] TJ ET BT 26.250 214.020 Td /F1 9.8 Tf [(SRR2566828, SRR2566830, SRR3086799- SRR3086806, SRR3086823, SRR3086828, SRR3086831, SRR3086834, )] TJ ET BT 26.250 202.115 Td /F1 9.8 Tf [(SRR3095255 \(see Table 1 for details\).)] TJ ET BT 26.250 165.512 Td /F6 12.0 Tf [(Competing Interest Statement)] TJ ET BT 26.250 145.558 Td /F1 9.8 Tf [(The authors have declared that no competing interests exist.)] TJ ET BT 26.250 108.956 Td /F6 12.0 Tf [(Corresponding Authors)] TJ ET BT 26.250 89.001 Td /F1 9.8 Tf [(Vishnu Chaturvedi: Vishnu.Chaturvedi@cdph.ca.gov)] TJ ET BT 26.250 69.597 Td /F1 9.8 Tf [(Varvara K. Kozyreva: Varvara.Kozyreva@cdph.ca.gov)] TJ ET Q q 15.000 22.564 577.500 754.436 re W n 0.965 0.965 0.965 rg 26.250 744.514 555.000 32.486 re f 0.267 0.267 0.267 rg 0.267 0.267 0.267 RG 26.250 744.514 m 581.250 744.514 l 581.250 745.264 l 26.250 745.264 l f q 35.250 755.764 537.000 21.236 re W n 0.271 0.267 0.267 rg BT 35.250 766.011 Td /F6 9.8 Tf [(Table 2.)] TJ ET BT 71.559 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 119.782 766.011 Td /F1 9.8 Tf [( Pathogenicity Islands in outbreak-related )] TJ ET BT 301.327 766.011 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 349.551 766.011 Td /F1 9.8 Tf [( Poona isolates.)] TJ ET Q BT 26.250 727.490 Td /F1 9.8 Tf [(All isolates possessed phage elements homologous to )] TJ ET BT 263.614 727.490 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 311.837 727.490 Td /F1 9.8 Tf [( phage Fels-1 \(NC_010391\) and partial )] TJ ET BT 483.623 727.490 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 547.027 727.490 Td /F1 9.8 Tf [( phage )] TJ ET BT 26.250 715.585 Td /F1 9.8 Tf [(P4 \(NC_001609\). Four isolates harbored the genes homologous to an additional bacteriophage of either of the following types: )] TJ ET BT 26.250 703.680 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 94.529 703.680 Td /F1 9.8 Tf [( phage S-SKS1 \(NC_020851\), )] TJ ET BT 228.387 703.680 Td /F5 9.8 Tf [(Ectocarpus )] TJ ET BT 279.867 703.680 Td /F5 9.8 Tf [(siliculosus)] TJ ET BT 324.288 703.680 Td /F1 9.8 Tf [( virus 1 \(NC_002687\), )] TJ ET BT 422.363 703.680 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 485.767 703.680 Td /F1 9.8 Tf [( phage ES18 )] TJ ET BT 26.250 691.776 Td /F1 9.8 Tf [(\(NC_006949\), or )] TJ ET BT 101.569 691.776 Td /F5 9.8 Tf [(Enterobacteria)] TJ ET BT 164.973 691.776 Td /F1 9.8 Tf [( phage P1 \(NC_005856\). Interestingly, phage genes found in one of the isolates were )] TJ ET BT 26.250 679.871 Td /F1 9.8 Tf [(homologous to S-SKS1 bacteriophage, which was described as a virus infecting )] TJ ET BT 373.613 679.871 Td /F5 9.8 Tf [(Synechococcus)] TJ ET BT 441.892 679.871 Td /F1 9.8 Tf [(, a genus of Cyanobacteria )] TJ ET BT 26.250 667.966 Td /F1 9.8 Tf [(known to be one of the predominant prokaryotic components of the oceans picophytoplankton)] TJ ET 0.267 0.267 0.267 rg BT 432.718 669.473 Td /F6 8.7 Tf [(12)] TJ ET 0.271 0.267 0.267 rg BT 442.355 667.966 Td /F1 9.8 Tf [(. Another isolate possessed )] TJ ET BT 26.250 656.061 Td /F1 9.8 Tf [(genes homologous to EsV-1 bacteriophage, characterized previously as a bacteriophage of the marine filamentous brown algae )] TJ ET BT 26.250 644.157 Td /F5 9.8 Tf [(Ectocarpus siliculosus)] TJ ET 0.267 0.267 0.267 rg BT 122.151 645.664 Td /F6 8.7 Tf [(13)] TJ ET 0.271 0.267 0.267 rg BT 131.788 644.157 Td /F1 9.8 Tf [(. The sequences identified as incomplete S-SKS1 or EsV-1 phages were integrated into the larger )] TJ ET BT 26.250 632.252 Td /F1 9.8 Tf [(contigs with clear )] TJ ET BT 103.733 632.252 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 151.957 632.252 Td /F1 9.8 Tf [( identity according to BLAST search. The marine phage-like sequences were also common in )] TJ ET BT 26.250 620.347 Td /F1 9.8 Tf [(other )] TJ ET BT 51.181 620.347 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 136.796 620.347 Td /F1 9.8 Tf [( serotypes found in NCBI database and seem to represent a part of the )] TJ ET BT 445.714 620.347 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 493.938 620.347 Td /F1 9.8 Tf [( accessory genome.)] TJ ET BT 26.250 600.942 Td /F1 9.8 Tf [(WGS has proven to be a powerful tool for the investigation of outbreaks caused by )] TJ ET BT 384.475 600.942 Td /F5 9.8 Tf [(Salmonella enterica)] TJ ET BT 470.089 600.942 Td /F1 9.8 Tf [(, providing great )] TJ ET BT 26.250 589.038 Td /F1 9.8 Tf [(epidemiological concordance and higher resolution than PFGE, the traditional method for )] TJ ET BT 412.642 589.038 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 460.866 589.038 Td /F1 9.8 Tf [( strain subtyping)] TJ ET 0.267 0.267 0.267 rg BT 531.856 590.545 Td /F6 8.7 Tf [(14)] TJ ET 0.271 0.267 0.267 rg BT 541.493 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 543.902 590.545 Td /F6 8.7 Tf [(15)] TJ ET 0.271 0.267 0.267 rg BT 553.540 592.926 Td /F1 8.7 Tf [(,)] TJ ET 0.267 0.267 0.267 rg BT 555.949 590.545 Td /F6 8.7 Tf [(16)] TJ ET 0.271 0.267 0.267 rg BT 565.586 589.038 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 577.133 Td /F1 9.8 Tf [(In our study, WGS confirmed the genetic relatedness of strains with similar PFGE patterns. Outbreak-related isolates exhibited )] TJ ET BT 26.250 565.228 Td /F1 9.8 Tf [(3 PFGE patterns which complicated assignment of the isolates to a single outbreak source. Particularly, isolate M15X04477 )] TJ ET BT 26.250 553.323 Td /F1 9.8 Tf [(with the pattern JL6X01.0778 differed by 3 bands from the predominant outbreak PFGE-type, thus introducing a higher degree )] TJ ET BT 26.250 541.419 Td /F1 9.8 Tf [(of uncertainty regarding the inclusion of this isolate in the outbreak cluster even though the epidemiologic information supported )] TJ ET BT 26.250 529.514 Td /F1 9.8 Tf [(a possible link to the outbreak. WGS, on the other hand, showed unambiguous clustering of all epidemiologically-related )] TJ ET BT 26.250 517.609 Td /F1 9.8 Tf [(isolates together and clarified genetic relatedness of M15X04477 to the other outbreak isolates. Two case-patients denying )] TJ ET BT 26.250 505.704 Td /F1 9.8 Tf [(cucumber consumption also clustered with the outbreak, which could be explained by possible secondary transmission, cross-)] TJ ET BT 26.250 493.800 Td /F1 9.8 Tf [(contamination of other foods, poor food history recall, or case-patients not noticing cucumbers as part of a multi-ingredient meal )] TJ ET BT 26.250 481.895 Td /F1 9.8 Tf [(such as a salad or a sandwich. In support of the high reproducibility of the method, the results of WGS performed by two )] TJ ET BT 26.250 469.990 Td /F1 9.8 Tf [(different laboratories were identical. Additionally, WGS provided information about the MLST sequence type and acquired )] TJ ET BT 26.250 458.085 Td /F1 9.8 Tf [(components of the genome, allowing for more comprehensive characterization of the isolates. Though MLST usually provides )] TJ ET BT 26.250 446.181 Td /F1 9.8 Tf [(poor resolution between )] TJ ET BT 133.012 446.181 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 181.236 446.181 Td /F1 9.8 Tf [( strains, it is helpful for understanding the global epidemiology of infectious clones)] TJ ET 0.267 0.267 0.267 rg BT 533.484 447.688 Td /F6 8.7 Tf [(17)] TJ ET 0.271 0.267 0.267 rg BT 543.121 446.181 Td /F1 9.8 Tf [(. )] TJ ET BT 26.250 434.276 Td /F1 9.8 Tf [(Diversity in accessory genes of the studied isolates demonstrates the plasticity of the )] TJ ET BT 394.741 434.276 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 442.965 434.276 Td /F1 9.8 Tf [( Poona genome which is )] TJ ET BT 26.250 422.371 Td /F1 9.8 Tf [(potentially responsible for variations in PFGE patterns that do not necessarily reflect the phylogenetic distance between the )] TJ ET BT 26.250 410.466 Td /F1 9.8 Tf [(isolates)] TJ ET 0.267 0.267 0.267 rg BT 59.303 411.974 Td /F6 8.7 Tf [(18)] TJ ET 0.271 0.267 0.267 rg BT 68.940 410.466 Td /F1 9.8 Tf [(. Since the loss/acquisition of mobile elements does not affect genome-wide SNPs calling, it explains higher )] TJ ET BT 26.250 398.562 Td /F1 9.8 Tf [(evolutionary congruence of WGS-inferred phylogeny compared with PFGE. Noteworthy, WGS results were acquired in 7 )] TJ ET BT 26.250 386.657 Td /F1 9.8 Tf [(working days from the time of pure culture reception by the laboratory. Our experience suggests that WGS can be used for )] TJ ET BT 26.250 374.752 Td /F1 9.8 Tf [(routine epidemiological subtyping.)] TJ ET BT 26.250 338.150 Td /F6 12.0 Tf [(Conclusions)] TJ ET BT 26.250 318.195 Td /F1 9.8 Tf [(Real-time PFGE and WGS genotyping results from this )] TJ ET BT 266.831 318.195 Td /F5 9.8 Tf [(Salmonella)] TJ ET BT 315.055 318.195 Td /F1 9.8 Tf [( Poona outbreak provided concordant assignment of isolates )] TJ ET BT 26.250 306.291 Td /F1 9.8 Tf [(to the outbreak cluster by both methods. However, the WGS allowed for a more unequivocal determination of the genetic )] TJ ET BT 26.250 294.386 Td /F1 9.8 Tf [(relatedness of the isolates than PFGE. WGS provided additional information regarding MLST type, pathogenicity genes, and )] TJ ET BT 26.250 282.481 Td /F1 9.8 Tf [(bacteriophage content of the isolates. WGS data obtained independently at two laboratories showed complete agreement.)] TJ ET BT 26.250 245.879 Td /F6 12.0 Tf [(Data Availability Statement)] TJ ET BT 26.250 225.924 Td /F1 9.8 Tf [(The raw reads for sequenced here samples were submitted to NCBI SRA archive under accession numbers SRR2566826- )] TJ ET BT 26.250 214.020 Td /F1 9.8 Tf [(SRR2566828, SRR2566830, SRR3086799- SRR3086806, SRR3086823, SRR3086828, SRR3086831, SRR3086834, )] TJ ET BT 26.250 202.115 Td /F1 9.8 Tf [(SRR3095255 \(see Table 1 for details\).)] TJ ET BT 26.250 165.512 Td /F6 12.0 Tf [(Competing Interest Statement)] TJ ET BT 26.250 145.558 Td /F1 9.8 Tf [(The authors have declared that no competing interests exist.)] TJ ET BT 26.250 108.956 Td /F6 12.0 Tf [(Corresponding Authors)] TJ ET BT 26.250 89.001 Td /F1 9.8 Tf [(Vishnu Chaturvedi: Vishnu.Chaturvedi@cdph.ca.gov)] TJ ET BT 26.250 69.597 Td /F1 9.8 Tf [(Varvara K. Kozyreva: Varvara.Kozyreva@cdph.ca.gov)] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(5)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 240 0 obj << /Type /Annot /Subtype /Link /A 241 0 R /Border [0 0 0] /H /I /Rect [ 432.7178 668.6717 442.3551 677.4900 ] >> endobj 241 0 obj << /Type /Action >> endobj 242 0 obj << /Type /Annot /Subtype /Link /A 243 0 R /Border [0 0 0] /H /I /Rect [ 122.1510 644.8622 131.7883 653.6805 ] >> endobj 243 0 obj << /Type /Action >> endobj 244 0 obj << /Type /Annot /Subtype /Link /A 245 0 R /Border [0 0 0] /H /I /Rect [ 531.8557 589.7432 541.4931 598.5615 ] >> endobj 245 0 obj << /Type /Action >> endobj 246 0 obj << /Type /Annot /Subtype /Link /A 247 0 R /Border [0 0 0] /H /I /Rect [ 543.9024 589.7432 553.5398 598.5615 ] >> endobj 247 0 obj << /Type /Action >> endobj 248 0 obj << /Type /Annot /Subtype /Link /A 249 0 R /Border [0 0 0] /H /I /Rect [ 555.9491 589.7432 565.5864 598.5615 ] >> endobj 249 0 obj << /Type /Action >> endobj 250 0 obj << /Type /Annot /Subtype /Link /A 251 0 R /Border [0 0 0] /H /I /Rect [ 533.4840 446.8862 543.1213 455.7045 ] >> endobj 251 0 obj << /Type /Action >> endobj 252 0 obj << /Type /Annot /Subtype /Link /A 253 0 R /Border [0 0 0] /H /I /Rect [ 59.3025 411.1719 68.9398 419.9902 ] >> endobj 253 0 obj << /Type /Action >> endobj 254 0 obj << /Type /Annot /Subtype /Link /A 255 0 R /Border [0 0 0] /H /I /Rect [ 432.7178 668.6717 442.3551 677.4900 ] >> endobj 255 0 obj << /Type /Action >> endobj 256 0 obj << /Type /Annot /Subtype /Link /A 257 0 R /Border [0 0 0] /H /I /Rect [ 122.1510 644.8622 131.7883 653.6805 ] >> endobj 257 0 obj << /Type /Action >> endobj 258 0 obj << /Type /Annot /Subtype /Link /A 259 0 R /Border [0 0 0] /H /I /Rect [ 531.8557 589.7432 541.4931 598.5615 ] >> endobj 259 0 obj << /Type /Action >> endobj 260 0 obj << /Type /Annot /Subtype /Link /A 261 0 R /Border [0 0 0] /H /I /Rect [ 543.9024 589.7432 553.5398 598.5615 ] >> endobj 261 0 obj << /Type /Action >> endobj 262 0 obj << /Type /Annot /Subtype /Link /A 263 0 R /Border [0 0 0] /H /I /Rect [ 555.9491 589.7432 565.5864 598.5615 ] >> endobj 263 0 obj << /Type /Action >> endobj 264 0 obj << /Type /Annot /Subtype /Link /A 265 0 R /Border [0 0 0] /H /I /Rect [ 533.4840 446.8862 543.1213 455.7045 ] >> endobj 265 0 obj << /Type /Action >> endobj 266 0 obj << /Type /Annot /Subtype /Link /A 267 0 R /Border [0 0 0] /H /I /Rect [ 59.3025 411.1719 68.9398 419.9902 ] >> endobj 267 0 obj << /Type /Action >> endobj 268 0 obj << /Type /Annot /Subtype /Link /A 269 0 R /Border [0 0 0] /H /I /Rect [ 432.7178 668.6717 442.3551 677.4900 ] >> endobj 269 0 obj << /Type /Action >> endobj 270 0 obj << /Type /Annot /Subtype /Link /A 271 0 R /Border [0 0 0] /H /I /Rect [ 122.1510 644.8622 131.7883 653.6805 ] >> endobj 271 0 obj << /Type /Action >> endobj 272 0 obj << /Type /Annot /Subtype /Link /A 273 0 R /Border [0 0 0] /H /I /Rect [ 531.8557 589.7432 541.4931 598.5615 ] >> endobj 273 0 obj << /Type /Action >> endobj 274 0 obj << /Type /Annot /Subtype /Link /A 275 0 R /Border [0 0 0] /H /I /Rect [ 543.9024 589.7432 553.5398 598.5615 ] >> endobj 275 0 obj << /Type /Action >> endobj 276 0 obj << /Type /Annot /Subtype /Link /A 277 0 R /Border [0 0 0] /H /I /Rect [ 555.9491 589.7432 565.5864 598.5615 ] >> endobj 277 0 obj << /Type /Action >> endobj 278 0 obj << /Type /Annot /Subtype /Link /A 279 0 R /Border [0 0 0] /H /I /Rect [ 533.4840 446.8862 543.1213 455.7045 ] >> endobj 279 0 obj << /Type /Action >> endobj 280 0 obj << /Type /Annot /Subtype /Link /A 281 0 R /Border [0 0 0] /H /I /Rect [ 59.3025 411.1719 68.9398 419.9902 ] >> endobj 281 0 obj << /Type /Action >> endobj 282 0 obj << /Type /Page /Parent 3 0 R /Contents 283 0 R >> endobj 283 0 obj << /Length 3901 >> stream 0.271 0.267 0.267 rg q 15.000 28.887 577.500 748.113 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(Acknowledgements)] TJ ET BT 26.250 730.324 Td /F1 9.8 Tf [(We acknowledge colleagues in Food & Waterborne Diseases and Immunodiagnostics & Parasitic Sections of CDPH Microbial )] TJ ET BT 26.250 718.419 Td /F1 9.8 Tf [(Disease Laboratory Branch, particularly Francine Arroyo, Katherine Trinh, Frank Ni, and Greg Inami for processing and )] TJ ET BT 26.250 706.515 Td /F1 9.8 Tf [(identification of bacterial cultures. We thank the CDPH Infectious Diseases Branch, and particularly Dr. Duc Vugia for the critical )] TJ ET BT 26.250 694.610 Td /F1 9.8 Tf [(review of this work. We acknowledge valuable assistance from the CDPH Food and Drug Branch \(Michael Needham, Amber )] TJ ET BT 26.250 682.705 Td /F1 9.8 Tf [(Barnes, Samantha Mauldin, Brandon Adcock\), and the California local health departments, particularly County of San Diego )] TJ ET BT 26.250 670.800 Td /F1 9.8 Tf [(Health and Human Services Agency \(Azarnoush Maroufi, Eric C. McDonald, Jessica Healy, Brian Murphy, Loreto N. Negado\) )] TJ ET BT 26.250 658.896 Td /F1 9.8 Tf [(and County of San Diego Department of Environmental Health \(Heather Buonomo, Cory Neiss\).)] TJ ET Q q 15.000 28.887 577.500 748.113 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(Acknowledgements)] TJ ET BT 26.250 730.324 Td /F1 9.8 Tf [(We acknowledge colleagues in Food & Waterborne Diseases and Immunodiagnostics & Parasitic Sections of CDPH Microbial )] TJ ET BT 26.250 718.419 Td /F1 9.8 Tf [(Disease Laboratory Branch, particularly Francine Arroyo, Katherine Trinh, Frank Ni, and Greg Inami for processing and )] TJ ET BT 26.250 706.515 Td /F1 9.8 Tf [(identification of bacterial cultures. We thank the CDPH Infectious Diseases Branch, and particularly Dr. Duc Vugia for the critical )] TJ ET BT 26.250 694.610 Td /F1 9.8 Tf [(review of this work. We acknowledge valuable assistance from the CDPH Food and Drug Branch \(Michael Needham, Amber )] TJ ET BT 26.250 682.705 Td /F1 9.8 Tf [(Barnes, Samantha Mauldin, Brandon Adcock\), and the California local health departments, particularly County of San Diego )] TJ ET BT 26.250 670.800 Td /F1 9.8 Tf [(Health and Human Services Agency \(Azarnoush Maroufi, Eric C. McDonald, Jessica Healy, Brian Murphy, Loreto N. Negado\) )] TJ ET BT 26.250 658.896 Td /F1 9.8 Tf [(and County of San Diego Department of Environmental Health \(Heather Buonomo, Cory Neiss\).)] TJ ET Q q 15.000 28.887 577.500 748.113 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(Acknowledgements)] TJ ET BT 26.250 730.324 Td /F1 9.8 Tf [(We acknowledge colleagues in Food & Waterborne Diseases and Immunodiagnostics & Parasitic Sections of CDPH Microbial )] TJ ET BT 26.250 718.419 Td /F1 9.8 Tf [(Disease Laboratory Branch, particularly Francine Arroyo, Katherine Trinh, Frank Ni, and Greg Inami for processing and )] TJ ET BT 26.250 706.515 Td /F1 9.8 Tf [(identification of bacterial cultures. We thank the CDPH Infectious Diseases Branch, and particularly Dr. Duc Vugia for the critical )] TJ ET BT 26.250 694.610 Td /F1 9.8 Tf [(review of this work. We acknowledge valuable assistance from the CDPH Food and Drug Branch \(Michael Needham, Amber )] TJ ET BT 26.250 682.705 Td /F1 9.8 Tf [(Barnes, Samantha Mauldin, Brandon Adcock\), and the California local health departments, particularly County of San Diego )] TJ ET BT 26.250 670.800 Td /F1 9.8 Tf [(Health and Human Services Agency \(Azarnoush Maroufi, Eric C. McDonald, Jessica Healy, Brian Murphy, Loreto N. Negado\) )] TJ ET BT 26.250 658.896 Td /F1 9.8 Tf [(and County of San Diego Department of Environmental Health \(Heather Buonomo, Cory Neiss\).)] TJ ET Q q 0.000 0.000 0.000 rg BT 291.710 19.825 Td /F1 11.0 Tf [(6)] TJ ET BT 25.000 19.825 Td /F1 11.0 Tf [(PLOS Currents Outbreaks)] TJ ET Q endstream endobj 284 0 obj << /Type /Page /Parent 3 0 R /Contents 285 0 R >> endobj 285 0 obj << /Length 18031 >> stream 0.271 0.267 0.267 rg q 15.000 149.372 577.500 627.628 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(References)] TJ ET BT 26.250 722.824 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 722.824 Td /F1 9.8 Tf [(Centers for Disease Control and Prevention. Multistate outbreaks of Salmonella serotype Poona infections associated with )] TJ ET BT 26.250 710.919 Td /F1 9.8 Tf [(eating cantaloupe from MexicoUnited States and Canada, 20002002. MMWR Morb Mortal Wkly Rep. 2002;51:10447.)] TJ ET BT 26.250 691.515 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 691.515 Td /F1 9.8 Tf [(Jackson BR, Griffin PM, Cole D, Walsh KA, Chai SJ: Outbreak-associated Salmonella enterica serotypes and food )] TJ ET BT 26.250 679.610 Td /F1 9.8 Tf [(Commodities, United States, 1998-2008. Emerging infectious diseases 2013, 19\(8\):1239-1244.)] TJ ET BT 26.250 660.205 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 660.205 Td /F1 9.8 Tf [(Walters MS, Simmons L, Anderson TC, DeMent J, Van Zile K, Matthias LP, Etheridge S, Baker R, Healan C, Bagby R et al: )] TJ ET BT 26.250 648.300 Td /F1 9.8 Tf [(Outbreaks of Salmonellosis From Small Turtles. Pediatrics 2016, 137\(1\).)] TJ ET BT 26.250 628.896 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 628.896 Td /F1 9.8 Tf [(Multistate Outbreak of Salmonella Poona Infections Linked to Imported Cucumbers \(Final Update\) )] TJ ET BT 26.250 616.991 Td /F1 9.8 Tf [([http://www.cdc.gov/salmonella/poona-09-15/index.html])] TJ ET BT 26.250 597.586 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 597.586 Td /F1 9.8 Tf [(Ribot EM, Fair MA, Gautom R, Cameron DN, Hunter SB, Swaminathan B, Barrett TJ: Standardization of pulsed-field gel )] TJ ET BT 26.250 585.681 Td /F1 9.8 Tf [(electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne )] TJ ET BT 26.250 573.777 Td /F1 9.8 Tf [(pathogens and disease 2006, 3\(1\):59-67.)] TJ ET BT 26.250 554.372 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 554.372 Td /F1 9.8 Tf [(Seemann T \(2014\) Prokka: rapid prokaryotic genome annotation. Bioinformatics 30: 2068-2069.)] TJ ET BT 26.250 534.967 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 534.967 Td /F1 9.8 Tf [(Joensen KG, Scheutz F, Lund O, Hasman H, Kaas RS, et al. \(2014\) Real-time whole-genome sequencing for routine typing, )] TJ ET BT 26.250 523.062 Td /F1 9.8 Tf [(surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol 52: 1501-1510.)] TJ ET BT 26.250 503.658 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 503.658 Td /F1 9.8 Tf [(Zhou Y, Liang Y, Lynch KH, Dennis JJ, Wishart DS \(2011\) PHAST: a fast phage search tool. Nucleic Acids Res 39: W347-)] TJ ET BT 26.250 491.753 Td /F1 9.8 Tf [(352.)] TJ ET BT 26.250 472.348 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 472.348 Td /F1 9.8 Tf [(Larsen MV, Cosentino S, Rasmussen S, Friis C, Hasman H, et al. \(2012\) Multilocus sequence typing of total-genome-)] TJ ET BT 26.250 460.443 Td /F1 9.8 Tf [(sequenced bacteria. J Clin Microbiol 50: 1355-1361.)] TJ ET BT 26.250 441.039 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 441.039 Td /F1 9.8 Tf [(Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, et al. \(2009\) The Sequence Alignment/Map format and SAMtools. )] TJ ET BT 26.250 429.134 Td /F1 9.8 Tf [(Bioinformatics 25: 2078-2079.)] TJ ET BT 26.250 409.729 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 409.729 Td /F1 9.8 Tf [(Danecek P, Auton A, Abecasis G, Albers CA, Banks E, et al. \(2011\) The variant call format and VCFtools. Bioinformatics )] TJ ET BT 26.250 397.824 Td /F1 9.8 Tf [(27: 2156-2158.)] TJ ET BT 26.250 378.420 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 378.420 Td /F1 9.8 Tf [(Mann NH \(2003\) Phages of the marine cyanobacterial picophytoplankton. FEMS Microbiol Rev 27: 17-34.)] TJ ET BT 26.250 359.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 359.015 Td /F1 9.8 Tf [(Delaroque N, Muller DG, Bothe G, Pohl T, Knippers R, et al. \(2001\) The complete DNA sequence of the Ectocarpus )] TJ ET BT 26.250 347.110 Td /F1 9.8 Tf [(siliculosus Virus EsV-1 genome. Virology 287: 112-132.)] TJ ET BT 26.250 327.705 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 327.705 Td /F1 9.8 Tf [(Deng X, Shariat N, Driebe EM, Roe CC, Tolar B, et al. \(2015\) Comparative analysis of subtyping methods against a whole-)] TJ ET BT 26.250 315.801 Td /F1 9.8 Tf [(genome-sequencing standard for Salmonella enterica serotype Enteritidis. J Clin Microbiol 53: 212-218.)] TJ ET BT 26.250 296.396 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 296.396 Td /F1 9.8 Tf [(Scaltriti E, Sassera D, Comandatore F, Morganti M, Mandalari C, et al. \(2015\) Differential single nucleotide polymorphism-)] TJ ET BT 26.250 284.491 Td /F1 9.8 Tf [(based analysis of an outbreak caused by Salmonella enterica serovar Manhattan reveals epidemiological details missed by )] TJ ET BT 26.250 272.586 Td /F1 9.8 Tf [(standard pulsed-field gel electrophoresis. J Clin Microbiol 53: 1227-1238.)] TJ ET BT 26.250 253.182 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 253.182 Td /F1 9.8 Tf [(Taylor AJ, Lappi V, Wolfgang WJ, Lapierre P, Palumbo MJ, et al. \(2015\) Characterization of Foodborne Outbreaks of )] TJ ET BT 26.250 241.277 Td /F1 9.8 Tf [(Salmonella enterica Serovar Enteritidis with Whole-Genome Sequencing Single Nucleotide Polymorphism-Based Analysis for )] TJ ET BT 26.250 229.372 Td /F1 9.8 Tf [(Surveillance and Outbreak Detection. J Clin Microbiol 53: 3334-3340.)] TJ ET BT 26.250 209.967 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 209.967 Td /F1 9.8 Tf [(Sabat AJ, Budimir A, Nashev D, Sa-Leao R, van Dijl J, et al. \(2013\) Overview of molecular typing methods for outbreak )] TJ ET BT 26.250 198.063 Td /F1 9.8 Tf [(detection and epidemiological surveillance. Euro Surveill 18: 20380.)] TJ ET BT 26.250 178.658 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 178.658 Td /F1 9.8 Tf [(Gurtler V, Mayall BC \(2001\) Genomic approaches to typing, taxonomy and evolution of bacterial isolates. Int J Syst Evol )] TJ ET BT 26.250 166.753 Td /F1 9.8 Tf [(Microbiol 51: 3-16.)] TJ ET Q q 15.000 149.372 577.500 627.628 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(References)] TJ ET BT 26.250 722.824 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 722.824 Td /F1 9.8 Tf [(Centers for Disease Control and Prevention. Multistate outbreaks of Salmonella serotype Poona infections associated with )] TJ ET BT 26.250 710.919 Td /F1 9.8 Tf [(eating cantaloupe from MexicoUnited States and Canada, 20002002. MMWR Morb Mortal Wkly Rep. 2002;51:10447.)] TJ ET BT 26.250 691.515 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 691.515 Td /F1 9.8 Tf [(Jackson BR, Griffin PM, Cole D, Walsh KA, Chai SJ: Outbreak-associated Salmonella enterica serotypes and food )] TJ ET BT 26.250 679.610 Td /F1 9.8 Tf [(Commodities, United States, 1998-2008. Emerging infectious diseases 2013, 19\(8\):1239-1244.)] TJ ET BT 26.250 660.205 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 660.205 Td /F1 9.8 Tf [(Walters MS, Simmons L, Anderson TC, DeMent J, Van Zile K, Matthias LP, Etheridge S, Baker R, Healan C, Bagby R et al: )] TJ ET BT 26.250 648.300 Td /F1 9.8 Tf [(Outbreaks of Salmonellosis From Small Turtles. Pediatrics 2016, 137\(1\).)] TJ ET BT 26.250 628.896 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 628.896 Td /F1 9.8 Tf [(Multistate Outbreak of Salmonella Poona Infections Linked to Imported Cucumbers \(Final Update\) )] TJ ET BT 26.250 616.991 Td /F1 9.8 Tf [([http://www.cdc.gov/salmonella/poona-09-15/index.html])] TJ ET BT 26.250 597.586 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 597.586 Td /F1 9.8 Tf [(Ribot EM, Fair MA, Gautom R, Cameron DN, Hunter SB, Swaminathan B, Barrett TJ: Standardization of pulsed-field gel )] TJ ET BT 26.250 585.681 Td /F1 9.8 Tf [(electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne )] TJ ET BT 26.250 573.777 Td /F1 9.8 Tf [(pathogens and disease 2006, 3\(1\):59-67.)] TJ ET BT 26.250 554.372 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 554.372 Td /F1 9.8 Tf [(Seemann T \(2014\) Prokka: rapid prokaryotic genome annotation. Bioinformatics 30: 2068-2069.)] TJ ET BT 26.250 534.967 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 534.967 Td /F1 9.8 Tf [(Joensen KG, Scheutz F, Lund O, Hasman H, Kaas RS, et al. \(2014\) Real-time whole-genome sequencing for routine typing, )] TJ ET BT 26.250 523.062 Td /F1 9.8 Tf [(surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol 52: 1501-1510.)] TJ ET BT 26.250 503.658 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 503.658 Td /F1 9.8 Tf [(Zhou Y, Liang Y, Lynch KH, Dennis JJ, Wishart DS \(2011\) PHAST: a fast phage search tool. Nucleic Acids Res 39: W347-)] TJ ET BT 26.250 491.753 Td /F1 9.8 Tf [(352.)] TJ ET BT 26.250 472.348 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 472.348 Td /F1 9.8 Tf [(Larsen MV, Cosentino S, Rasmussen S, Friis C, Hasman H, et al. \(2012\) Multilocus sequence typing of total-genome-)] TJ ET BT 26.250 460.443 Td /F1 9.8 Tf [(sequenced bacteria. J Clin Microbiol 50: 1355-1361.)] TJ ET BT 26.250 441.039 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 441.039 Td /F1 9.8 Tf [(Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, et al. \(2009\) The Sequence Alignment/Map format and SAMtools. )] TJ ET BT 26.250 429.134 Td /F1 9.8 Tf [(Bioinformatics 25: 2078-2079.)] TJ ET BT 26.250 409.729 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 409.729 Td /F1 9.8 Tf [(Danecek P, Auton A, Abecasis G, Albers CA, Banks E, et al. \(2011\) The variant call format and VCFtools. Bioinformatics )] TJ ET BT 26.250 397.824 Td /F1 9.8 Tf [(27: 2156-2158.)] TJ ET BT 26.250 378.420 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 378.420 Td /F1 9.8 Tf [(Mann NH \(2003\) Phages of the marine cyanobacterial picophytoplankton. FEMS Microbiol Rev 27: 17-34.)] TJ ET BT 26.250 359.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 359.015 Td /F1 9.8 Tf [(Delaroque N, Muller DG, Bothe G, Pohl T, Knippers R, et al. \(2001\) The complete DNA sequence of the Ectocarpus )] TJ ET BT 26.250 347.110 Td /F1 9.8 Tf [(siliculosus Virus EsV-1 genome. Virology 287: 112-132.)] TJ ET BT 26.250 327.705 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 327.705 Td /F1 9.8 Tf [(Deng X, Shariat N, Driebe EM, Roe CC, Tolar B, et al. \(2015\) Comparative analysis of subtyping methods against a whole-)] TJ ET BT 26.250 315.801 Td /F1 9.8 Tf [(genome-sequencing standard for Salmonella enterica serotype Enteritidis. J Clin Microbiol 53: 212-218.)] TJ ET BT 26.250 296.396 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 296.396 Td /F1 9.8 Tf [(Scaltriti E, Sassera D, Comandatore F, Morganti M, Mandalari C, et al. \(2015\) Differential single nucleotide polymorphism-)] TJ ET BT 26.250 284.491 Td /F1 9.8 Tf [(based analysis of an outbreak caused by Salmonella enterica serovar Manhattan reveals epidemiological details missed by )] TJ ET BT 26.250 272.586 Td /F1 9.8 Tf [(standard pulsed-field gel electrophoresis. J Clin Microbiol 53: 1227-1238.)] TJ ET BT 26.250 253.182 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 253.182 Td /F1 9.8 Tf [(Taylor AJ, Lappi V, Wolfgang WJ, Lapierre P, Palumbo MJ, et al. \(2015\) Characterization of Foodborne Outbreaks of )] TJ ET BT 26.250 241.277 Td /F1 9.8 Tf [(Salmonella enterica Serovar Enteritidis with Whole-Genome Sequencing Single Nucleotide Polymorphism-Based Analysis for )] TJ ET BT 26.250 229.372 Td /F1 9.8 Tf [(Surveillance and Outbreak Detection. J Clin Microbiol 53: 3334-3340.)] TJ ET BT 26.250 209.967 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 209.967 Td /F1 9.8 Tf [(Sabat AJ, Budimir A, Nashev D, Sa-Leao R, van Dijl J, et al. \(2013\) Overview of molecular typing methods for outbreak )] TJ ET BT 26.250 198.063 Td /F1 9.8 Tf [(detection and epidemiological surveillance. Euro Surveill 18: 20380.)] TJ ET BT 26.250 178.658 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 178.658 Td /F1 9.8 Tf [(Gurtler V, Mayall BC \(2001\) Genomic approaches to typing, taxonomy and evolution of bacterial isolates. Int J Syst Evol )] TJ ET BT 26.250 166.753 Td /F1 9.8 Tf [(Microbiol 51: 3-16.)] TJ ET Q q 15.000 149.372 577.500 627.628 re W n 0.271 0.267 0.267 rg BT 26.250 750.278 Td /F6 12.0 Tf [(References)] TJ ET BT 26.250 722.824 Td /F1 9.8 Tf [(1.)] TJ ET BT 38.132 722.824 Td /F1 9.8 Tf [(Centers for Disease Control and Prevention. Multistate outbreaks of Salmonella serotype Poona infections associated with )] TJ ET BT 26.250 710.919 Td /F1 9.8 Tf [(eating cantaloupe from MexicoUnited States and Canada, 20002002. MMWR Morb Mortal Wkly Rep. 2002;51:10447.)] TJ ET BT 26.250 691.515 Td /F1 9.8 Tf [(2.)] TJ ET BT 38.132 691.515 Td /F1 9.8 Tf [(Jackson BR, Griffin PM, Cole D, Walsh KA, Chai SJ: Outbreak-associated Salmonella enterica serotypes and food )] TJ ET BT 26.250 679.610 Td /F1 9.8 Tf [(Commodities, United States, 1998-2008. Emerging infectious diseases 2013, 19\(8\):1239-1244.)] TJ ET BT 26.250 660.205 Td /F1 9.8 Tf [(3.)] TJ ET BT 38.132 660.205 Td /F1 9.8 Tf [(Walters MS, Simmons L, Anderson TC, DeMent J, Van Zile K, Matthias LP, Etheridge S, Baker R, Healan C, Bagby R et al: )] TJ ET BT 26.250 648.300 Td /F1 9.8 Tf [(Outbreaks of Salmonellosis From Small Turtles. Pediatrics 2016, 137\(1\).)] TJ ET BT 26.250 628.896 Td /F1 9.8 Tf [(4.)] TJ ET BT 38.132 628.896 Td /F1 9.8 Tf [(Multistate Outbreak of Salmonella Poona Infections Linked to Imported Cucumbers \(Final Update\) )] TJ ET BT 26.250 616.991 Td /F1 9.8 Tf [([http://www.cdc.gov/salmonella/poona-09-15/index.html])] TJ ET BT 26.250 597.586 Td /F1 9.8 Tf [(5.)] TJ ET BT 38.132 597.586 Td /F1 9.8 Tf [(Ribot EM, Fair MA, Gautom R, Cameron DN, Hunter SB, Swaminathan B, Barrett TJ: Standardization of pulsed-field gel )] TJ ET BT 26.250 585.681 Td /F1 9.8 Tf [(electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne )] TJ ET BT 26.250 573.777 Td /F1 9.8 Tf [(pathogens and disease 2006, 3\(1\):59-67.)] TJ ET BT 26.250 554.372 Td /F1 9.8 Tf [(6.)] TJ ET BT 38.132 554.372 Td /F1 9.8 Tf [(Seemann T \(2014\) Prokka: rapid prokaryotic genome annotation. Bioinformatics 30: 2068-2069.)] TJ ET BT 26.250 534.967 Td /F1 9.8 Tf [(7.)] TJ ET BT 38.132 534.967 Td /F1 9.8 Tf [(Joensen KG, Scheutz F, Lund O, Hasman H, Kaas RS, et al. \(2014\) Real-time whole-genome sequencing for routine typing, )] TJ ET BT 26.250 523.062 Td /F1 9.8 Tf [(surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol 52: 1501-1510.)] TJ ET BT 26.250 503.658 Td /F1 9.8 Tf [(8.)] TJ ET BT 38.132 503.658 Td /F1 9.8 Tf [(Zhou Y, Liang Y, Lynch KH, Dennis JJ, Wishart DS \(2011\) PHAST: a fast phage search tool. Nucleic Acids Res 39: W347-)] TJ ET BT 26.250 491.753 Td /F1 9.8 Tf [(352.)] TJ ET BT 26.250 472.348 Td /F1 9.8 Tf [(9.)] TJ ET BT 38.132 472.348 Td /F1 9.8 Tf [(Larsen MV, Cosentino S, Rasmussen S, Friis C, Hasman H, et al. \(2012\) Multilocus sequence typing of total-genome-)] TJ ET BT 26.250 460.443 Td /F1 9.8 Tf [(sequenced bacteria. J Clin Microbiol 50: 1355-1361.)] TJ ET BT 26.250 441.039 Td /F1 9.8 Tf [(10.)] TJ ET BT 43.553 441.039 Td /F1 9.8 Tf [(Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, et al. \(2009\) The Sequence Alignment/Map format and SAMtools. )] TJ ET BT 26.250 429.134 Td /F1 9.8 Tf [(Bioinformatics 25: 2078-2079.)] TJ ET BT 26.250 409.729 Td /F1 9.8 Tf [(11.)] TJ ET BT 43.553 409.729 Td /F1 9.8 Tf [(Danecek P, Auton A, Abecasis G, Albers CA, Banks E, et al. \(2011\) The variant call format and VCFtools. Bioinformatics )] TJ ET BT 26.250 397.824 Td /F1 9.8 Tf [(27: 2156-2158.)] TJ ET BT 26.250 378.420 Td /F1 9.8 Tf [(12.)] TJ ET BT 43.553 378.420 Td /F1 9.8 Tf [(Mann NH \(2003\) Phages of the marine cyanobacterial picophytoplankton. FEMS Microbiol Rev 27: 17-34.)] TJ ET BT 26.250 359.015 Td /F1 9.8 Tf [(13.)] TJ ET BT 43.553 359.015 Td /F1 9.8 Tf [(Delaroque N, Muller DG, Bothe G, Pohl T, Knippers R, et al. \(2001\) The complete DNA sequence of the Ectocarpus )] TJ ET BT 26.250 347.110 Td /F1 9.8 Tf [(siliculosus Virus EsV-1 genome. Virology 287: 112-132.)] TJ ET BT 26.250 327.705 Td /F1 9.8 Tf [(14.)] TJ ET BT 43.553 327.705 Td /F1 9.8 Tf [(Deng X, Shariat N, Driebe EM, Roe CC, Tolar B, et al. \(2015\) Comparative analysis of subtyping methods against a whole-)] TJ ET BT 26.250 315.801 Td /F1 9.8 Tf [(genome-sequencing standard for Salmonella enterica serotype Enteritidis. J Clin Microbiol 53: 212-218.)] TJ ET BT 26.250 296.396 Td /F1 9.8 Tf [(15.)] TJ ET BT 43.553 296.396 Td /F1 9.8 Tf [(Scaltriti E, Sassera D, Comandatore F, Morganti M, Mandalari C, et al. \(2015\) Differential single nucleotide polymorphism-)] TJ ET BT 26.250 284.491 Td /F1 9.8 Tf [(based analysis of an outbreak caused by Salmonella enterica serovar Manhattan reveals epidemiological details missed by )] TJ ET BT 26.250 272.586 Td /F1 9.8 Tf [(standard pulsed-field gel electrophoresis. J Clin Microbiol 53: 1227-1238.)] TJ ET BT 26.250 253.182 Td /F1 9.8 Tf [(16.)] TJ ET BT 43.553 253.182 Td /F1 9.8 Tf [(Taylor AJ, Lappi V, Wolfgang WJ, Lapierre P, Palumbo MJ, et al. \(2015\) Characterization of Foodborne Outbreaks of )] TJ ET BT 26.250 241.277 Td /F1 9.8 Tf [(Salmonella enterica Serovar Enteritidis with Whole-Genome Sequencing Single Nucleotide Polymorphism-Based Analysis for )] TJ ET BT 26.250 229.372 Td /F1 9.8 Tf [(Surveillance and Outbreak Detection. J Clin Microbiol 53: 3334-3340.)] TJ ET BT 26.250 209.967 Td /F1 9.8 Tf [(17.)] TJ ET BT 43.553 209.967 Td /F1 9.8 Tf [(Sabat AJ, Budimir A, Nashev D, Sa-Leao R, van Dijl J, et al. \(2013\) Overview of molecular typing methods for outbreak )] TJ ET BT 26.250 198.063 Td /F1 9.8 Tf [(detection and epidemiological surveillance. Euro Surveill 18: 20380.)] TJ ET BT 26.250 178.658 Td /F1 9.8 Tf [(18.)] TJ ET BT 43.553 178.658 Td /F1 9.8 Tf [(Gurtler V, Mayall BC \(2001\) Genomic approaches to typing, taxonomy and evolution of bacterial isolates. 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