Loïc Decrey a and Tamar Kohn a *

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1 Electronic Supplementry Mteril (ESI) for Environmentl Science: Wter Reserch & Technology. This journl is The Royl Society of Chemistry 2017 Supplementry mteril for Virus inctivtion in stored humn urine, sludge nd niml mnure under typicl conditions of storge or mesophilic nerobic digestion Loïc Decrey nd Tmr Kohn * Lbortory of Environmentl Chemistry, School of Architecture, Civil nd Environmentl Engineering (ENAC), École Polytechnique Fédérle de Lusnne (EPFL), CH-1015 Lusnne, Switzerlnd *Corresponding uthor phone: +41 (0) ; fx: +41 (0) ; e-mil: tmr.kohn@epfl.ch

2 Determintion of pk (s function of T) For ll species (except phosphte) (ccording to PHREEQC empiricl pk determintion) log 10 K = A 1 + A 2 T + A 3 + A T 4log 10 T + A 5 T 2 (S1) With: A1 A2 A3 A4 A5 CO HCO NH Kw And for OH - (K = [OH][H]/[H 2O] = K w/55.5): pk (T) = log 10 ( K w (T) 55.5 ) For phosphte (Vn t Hoff eqution): pk 2 = log 10 (10 pk 1 exp ( H R ( 1 T 2 1 T 1 ))) (S2) With: H r (stndrd enthlpy of rection) t 25 C: PO4/HPO kj/mol 1 HPO4/H2PO4-4.2 kj/mol And H r = H f (product)- H f (rectnt) (from online hndbook of chemistry nd physics): PO4 HPO4 H2PO kj/mol kj/mol kj/mol 1 Confirmed by PHREEQC dtbse (phreeqc.dt) kcl/mol = kj/mol

3 Scheme S1: Illustrtion of experimentl protocol

4 Tble S1: Composition of mtrixes used in this study. For those mtrices nlyzed t the beginning nd end of n experiment, the verge vlues re listed. 95% CI indictes the 95% confidence intervl of three (sludge nd mnure) or four (urine) replicte mesurements. SCOD = soluble chemicl oxygen demnd; TAN = totl mmonium nitrogen; TIC = totl inorgnic crbon. ID Smple description C Cl SCOD K Mg N TAN PO 4 TIC SO 4 [mm] 95% CI [mm] 95% CI [mgo 2 /L]95% CI [mm] 95% CI [mm] 95% CI [mm] 95% CI [mm] 95% CI [mm] 95% CI [mm] 95% CI [mm] 95% CI U1 Urine CH mle (2012) U U U U U U U U U U U12 CH mle (2013) < U13 CH mle (2014) < U14 CH femle U15 SA mix S1 Sludge CH Synthetic S2 CH Septic tnk S3 CH Synthetic M1 Mnure CH Pig M2 CH M3 CH Cow M4 CH Only one mesurement

5 k obs k pred T sensistivity ph sensistivity Ions mesured Tble S2: Mesured (k obs) nd predicted (k pred) inctivtion rte constnts. -1 C +1 C only TIC & NH4 Only NH4 Virus [dy -1 ] IC 95% R 2 [dy -1 ] [dy -1 ] [dy -1 ] [dy -1 ] [dy -1 ] [dy -1 ] [dy -1 ] MS2 U U U U U U U U U U U U U U U S S S M M M M HAdV U U S M T4 U S M (fst) (slow) PhiX174 U S M (fst) (slow) kobs determined with two dt points only

6 Clculted with PHREEQC Mesured Tble S3: Solution composition nd predicted inctivtion rte constnts for sludge nd mnure t the beginning nd end of the kinetic experiments. Sludge Mnure Smple ID S2 S3 M1 M2 M3 initil finl initil finl initil finl initil finl initil finl 0 dy 14 dys 0 dy 21 dys 0 dy 10 dys 0 dy 4 dys 0 dy 2 dys Avg. std Avg. std Avg. std Avg. std Avg. std Avg. std Avg. std Avg. std Avg. std Avg. std TIC [mm] TAN [mm] Cl [mm] SO4 [mm] PO4 [mm] C [mm] < Mg [mm] < N [mm] K [mm] SCOD [mgo2/l] ph [-] NH3 [mm] CO3 [mm] HCO3 [mm] OH [mm] PO4 [mm] 1.78E E E E E E E E E E-07 HPO4 [mm] k pred b Only one mesurement [dy -1 ] b Clculted with the model described in section 3.2

7 Figure S1: Bronsted plot of the pk of vrious bses versus their second-order rte constnt for inctivtion of MS2. kj ws determined, ccording to eq. 4, in queous solutions of the pure bse solution t 35 C (circle) nd in phosphtecrbonte (PC) nd mmonium-crbonte (AmC) buffer solutions for bses OH nd NH 3 respectively t 35 C (tringle) nd 20 C (squre). All dt re derived from Decrey et l. 1. pk vlues were corrected for temperture (see bove, determintion of pk).

8 Figure S2: kinetics of MS2 inctivtion in stored urine (U1-U15), sludge (S1-S3) nd mnure (M1-M4).

9 Figure S3: kinetics of ΦX174, T4 nd HAdV inctivtion in stored urine (U4, U12), sludge (S2) nd mnure (M4).

10 Figure S4: A. Accurcy of the predicted MS2 inctivtion rte constnt in stored urine (yellow tringles), sludge (brown squres) nd mnure (green dimonds). B. Sensitivity of the MS2 model prediction to temperture chnge (red), ph chnge (green) nd TIC nd TAN concentrtions (blue) in stored urine, sludge nd mnure. The prediction corresponding to the mesured properties of stored urine, sludge nd mnure (Tbles 1 nd S1) is represented in grey. Dt re shown in boxplot formt, where the thick line indictes the medin, the box is bounded by the first nd third qurtiles (50% of the dt) nd the whiskers indictes the minimum nd mximum of the rtio k pred/k obs (in percentge) determined for ll 22 smples tested.

11 Figure S5: Comprison of mesured nd predicted inctivtion rte constnts for MS2 t 35 C in 1:9 diluted urine without NH 4Cl ddition, nd with 35 nd 96 mmol L -1 NH 4Cl. NOH (9.8 nd 22.1 mmol L -1 ) ws dded with NH 4Cl to prtly compenste for ph drop. Finl ph vlues re: 8.13, 8.52 nd 8.46 from lowest to highest mmoni content. The solid line represents the 1:1 rtio between mesurement nd prediction.

12 Figure S6: detiled kinetics of ΦX174 nd T4 inctivtion in buffer solution (dt from Decrey et l. 2 ), stored urine (U12), sludge (S2) nd mnure (M4). The insets summrize the inctivtion rte constnts for the initil fst nd lte slow phse of the inctivtion curves in mnure. ph nd NH 3 ctivities of the different solutions re described in the legend.

13 References 1 L. Decrey, S. Kzm, K. M. Udert nd T. Kohn, Environ. Sci. Technol., 2015, 49, L. Decrey, S. Kzm nd T. Kohn, Appl. Environ. Microbiol., 2016, 82,

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