Rishindra M. Reddy, MD, William B

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Increased Variance in Oral and Gastric Microbiome Correlates With Esophagectomy Anastomotic Leak Rishindra M. Reddy, MD, William B. Weir, MD, Shari Barnett, RTT, BS, Brendan T. Heiden, BS, Mark B. Orringer, MD, Jules Lin, MD, Andrew C. Chang, MD, Philip W. Carrott, MD, William R. Lynch, MD, David G. Beer, PhD, J. Christopher Fenno, PhD, Yvonne Kapila, DDS, PhD The Annals of Thoracic Surgery Volume 105, Issue 3, Pages 865-870 (March 2018) DOI: 10.1016/j.athoracsur.2017.08.061 Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Schema of sampling during perioperative period for oral, wound, esophageal (Eso), and gastric specimens. The Annals of Thoracic Surgery 2018 105, 865-870DOI: (10.1016/j.athoracsur.2017.08.061) Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 (A–C) Community composition data. Shown is each sample (x-axis), with the number representing the specific patient, and the letter corresponding to the type of sample (A = preoperative saliva sample; B = esophageal sample; C = gastric sample; D = saliva sample taken 1 to 3 days after the operation; E = swab of cervical anastomotic leak; F = induced sputum sample). The legend shows the genus of the different microbiota identified through their 16S ribosomal RNA. The bars represent the percentage of each bacteria genus in each specimen. The Annals of Thoracic Surgery 2018 105, 865-870DOI: (10.1016/j.athoracsur.2017.08.061) Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Graph shows the microbiome diversity in each sample from principal coordinate analysis (PCA). The greater distance between two points reflects greater diversity in the microbiota in each sample. All specimens are reflected in this graph, with different colors representing different sources. Saliva samples are red preoperatively and orange postoperatively. Mucosal samples are green (esophageal) and purple (gastric). The arrows reflect the primary genus/operational taxonomic unit (OTU) at that specific area on the PCA. One patient had 2 preoperative saliva samples, with the secondary sample labeled in dark green. The axes represent the eigenvalues, which reflect the percentage that each axis contributes to the variability between data points. Axis 1 and axis 2 are the two largest eigenvalues for this analysis. (intraop = intraoperative; postop = postoperative; preop = preoperative.) The Annals of Thoracic Surgery 2018 105, 865-870DOI: (10.1016/j.athoracsur.2017.08.061) Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Patients with and without leaks were separated, and the data points from the preoperative saliva samples and intraoperative gastric samples are shown. Each red dot is paired to a blue dot from the same patient. Increased variance between oral and gastric microbiomes in patients with leak was noted when compared with the variance seen in patients without leaks (p < 0.015). The yellow dots denote the microbiome found on the neck swabs from patients with leaks. Not all leak swab specimens had measurable 16S ribosomal RNA. The axes represent the eigenvalues, which reflect the percentage that each axis contributes to the variability between data points. The Annals of Thoracic Surgery 2018 105, 865-870DOI: (10.1016/j.athoracsur.2017.08.061) Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions