## 5 Conclusions  
In summary, the present study provides new insights into the role of miRNAs as biomarkers of sepsis and resuscitation. We report, for the first time, changes in the serum expression of several miRNAs in a large animal model and their relationship with the intensity of fluid resuscitation. More studies comparing sepsis with non-infectious SIRS are needed to confirm our findings.  
The following are available online at [[https://www.mdpi.com/article/10.3390/app112311549/s1](https://www.mdpi.com/article/10.3390/app112311549/s1)]([https://www.mdpi.com/article/10.3390/app112311549/s1](https://www.mdpi.com/article/10.3390/app112311549/s1)). Table S1: Effect of sepsis and resuscitation on hemodynamic variables. Table S2: Effect of sepsis and resuscitation on biochemical variables. Table S3: Sample size for cytokine and miRNA determinations in the different groups. Table S4: Effect of sepsis and resuscitation on cytokine concentration. Table S5: Effect of sepsis and resuscitation on serum miRNA expression. Table S6: miRNAs differentially expressed in control versus septic animals: miRNA146-5p and miRNA34a-5p. Table S7: Pathways targeted by both miRNA34a-5p and miR-146a-5p (KEGG enrichment analysis) (adjusted \(p<0.01\)). Table S8: Proteins in our network participating in the Toll-like receptor and in the toxoplasmosis pathways. In red are proteins common to both pathways. Table S9: Enrichment analysis (KEGG) of proteins in the Toll-like receptor pathway (STRING) (adjusted \(p\) value \(<0.0000001\)). Table S10: Enrichment analysis (Gene Ontology biological process) of proteins in the Toll-like receptor pathway (STRING) (adjusted \(p\) value \(<0.0000001\)). Table S11: Enrichment analysis (reactome) of proteins in the Toll-like receptor pathway (STRING) (adjusted \(p\) value \(<0.001\)). Table S12: Enrichment analysis (KEGG) of proteins in the toxoplasmosis pathway (STRING) (adjusted \(p\) value \(<0.0000001\)). Table S13: Enrichment analysis (Gene Ontology biological process) of proteins in the toxoplasmosis pathway (STRING) (adjusted \(p\) value \(<0.0000001\)). Table S14: Enrichment analysis (reactome) of proteins in the toxoplasmosis pathway (STRING) (adjusted \(p\) value \(<0.0000001\)). Table S14: Enrichment analysis (reactome) of proteins in the toxoplasmosis pathway (STRING) (adjusted \(p\) value \(<0.001\)). Figure S1: Mortality of the different experimental groups (control, \(n=15\); sepsis 4 mL/kg/h, \(n=8\); sepsis 10 mL/kg/h, sepsis 17 mL/kg/h). Figure S2: (A) Genes targeted by both miRNAs of interest (miRNA146a-5p and miRNA34a-5p) (mirNet), where 1386 genes were identified as regulated byboth miRNAs. (B) Toxoplasma as the most highly significant pathway regulated by the miRNAs of interest (KEGG enrichment analysis) with 24 hits (adjusted \(p\) value = 3.77 \(\times\) 10\({}^{-5}\)), of which 10 were common with the Toll-like receptor signaling pathway. (C) Protein-protein interaction of the 24 proteins (STRING, confidence 0.9, in 4 clusters). PPI enrichment \(p\) value = 1.0 \(\times\) 10\({}^{-16}\). Purple line = experimental evidence; light blue line = database evidence; dotted lines = interactions appearing only after clustering.  
Conceptualization, L.O., A.F., N.N., R.H. and J.A.L.; methodology, A.F., R.H., R.P., M.A., M.d.P., L.M. and F.P.-V.; software, A.F., R.P. and P.G.-R.; validation, R.H., O.P., F.P.-V. and J.A.L.; formal analysis, N.N., L.M., R.H. and J.A.L.; investigation, L.O., A.F., N.N., M.A., F.P.-V. and J.A.L.; resources, F.P.-V. and J.A.L. All authors have read and agreed to the published version of the manuscript.  
The APC were waived by the editorial office.  
The Institutional Review Board StatementThe Institutional Review Board of Hospital Universitario de Getafe, approved on 24 September 2015 with the protocol number PI15/1942.  
Not applicable.  
Original data are available upon request.  
Not applicable.  
The authors declare no conflict of interest.