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Supplementary Information: Free Chlorine and Monochloramine Inactivation Kinetics of Aspergillus and Penicillium in Drinking Water
Xiao Ma1, Kyle Bibby1, 2*
1Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, PA 15261, 2Department of Computational and Systems Biology, University of Pittsburgh Medical School, Pittsburgh, PA 15261, USA *Corresponding author
*Corresponding Author: Kyle Bibby, 709 Benedum Hall, Pittsburgh, PA 15261 [email protected], 412-624-9207
Keywords:Disinfection, fungi, Aspergillus, Penicillium, opportunistic pathogen, premise plumbing
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WinBUGS code for delayed Chick-Watson model fitting
This code was derived from previous study (Sivaganesan et al. 2003)
Model
{
for (i in 1:N) {
Y[i] ~ dnorm(mu[i], tau)
mu[i] <- beta * (CT[i] - CTlag) * step(CT[i] - CTlag)
}
#non_informative priors (variance sigma2 = 1/tau, CTobs is the CT value corresponding to 10% of lowest Ln(N/N0))
tau ~ dgamma(0.001, 0.001)
beta ~ dnorm(0.0, 1.0E-6)
CTlag ~ dunif(0, CTobs)
sigma2 <- 1/tau
}
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0.8
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1.2
0 20 40 60 80
Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus type strain, 1 mg/L experiment
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0 10 20 30 40
Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus type strain, 4 mg/L experiment
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0.4
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0.8
1
0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus clinical strain, 1 mg/L exeperiment
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0.2
0.4
0.6
0.8
1
0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. versicolor, 1 mg/L exeperiment
0
0.2
0.4
0.6
0.8
1
0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
P. purpurogenum, 1 mg/L exeperiment
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11.5
22.5
33.5
4
0 10 20 30 40Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus clinical strain, 4 mg/L experiment
SI-Figure 1. Measured free chlorine residual concentration (mg/L)
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0.2
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0.6
0.8
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1.2
0 20 40 60 80
Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus type strain, 1 mg/L experiment
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4
0 10 20 30 40
Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus type strain, 4 mg/L experiment
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0.2
0.4
0.6
0.8
1
1.2
0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus clinical strain, 1 mg/L exeperiment
00.5
11.5
22.5
33.5
4
0 10 20 30 40Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. fumigatus clinical strain, 4 mg/L experiment
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1
0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
A. versicolor, 1 mg/L exeperiment
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0 20 40 60 80Free
chl
orin
e co
ncen
trat
ion
(mg/
L)
Time (min)
P. purpurogenum, 1 mg/L exeperiment
SI-Figure 2. Measured monochloramine residual concentration (mg/L)
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-1
-0.5
0
0.5
1
0 20 40 60
Ln(N
/N0)
Time (min)
Free chlorine control experiment
A. fumgiatus typestrain
A. fumigatus clinicalstrain
Aspergillus spp.
Penicillium spp.
-1
-0.5
0
0.5
1
0 20 40 60
Ln(N
/N0)
Time (min)
Monochloramine control experiment
A. fumigatus typestrain
A. fumigatus clinicalstrain
A. versicolor tapwater isolate
P. purpurogenumtap water isolate
A. fumigatus type strain
A. fumigatus clinical strain
A. versicolor
P. purpurogenum
SI-Figure 3. Measured mean Ln(N/N0) for free chlorine and monochloramine control experiments, which no disinfectant was used;
Further F-test results (conducted using Prism, GraphPad Software) suggested that the measured Ln(N/N0) of all tested strains did not significantly deviated from 0
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y = 0.97xR² = 0.95
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-6
-4
-2
0-8 -6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.87xR² = 0.75
-12
-10
-8
-6
-4
-2
0-12 -10 -8 -6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.93xR² = 0.87
-3
-2
-1
0-3 -2 -1 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.96xR² = 0.93
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-4
-2
0-6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.91xR² = 0.80
-8
-6
-4
-2
0-8 -6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.81xR² = 0.56
-12
-10
-8
-6
-4
-2
0-12 -10 -8 -6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.93xR² = 0.86
-3
-2
-1
0-3 -2 -1 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.96xR² = 0.91
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-3
-2
-1
0-6 -4 -2 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
Classical Chick-Watson Model
Delayed Chick-Watson Model
A. fumigatus type strain
A. fumigatus clinical strain
A. versicolortap water isolate
P. purpurogenumtap water isolate
y = 0.97xR² = 0.95
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-6
-4
-2
0-10 -5 0
Pred
icte
d …
Measured …
1 mg/L assay
4 mg/L assay
SI-Figure 4. Comparison of model predicted Ln(N/N0) against measured Ln(N/N0) for both delayed Chick-Watson model and classical Chick-Watson model, free chlorine disinfection
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y = 0.99xR² = 0.97
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-3
-1
-6 -4 -2 0
Mea
sure
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.94xR² = 0.87
-8
-6
-4
-2
0-8 -6 -4 -2 0
Mea
sure
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.98xR² = 0.97
-3
-2
-1
0-3 -2 -1 0
Mea
sure
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.87xR² = 0.75
-1
-0.5
0-1 -0.5 0
Pred
icte
d Ln
(N/N
0)
Measured Ln(N/N0)
y = 0.98xR² = 0.95
-6
-4
-2
0-6 -4 -2 0
Estim
ated
Ln(
N/N
0)
Measured Ln(N/N0)
y = 0.94xR² = 0.88
-6
-4
-2
0-6 -4 -2 0
Estim
ated
Ln(
N/N
0)
Measured Ln(N/N0)
y = 0.93xR² = 0.86
-3
-2
-1
0-3 -2 -1 0
Estim
ated
Ln(
N/N
0)
Measured Ln(N/N0)
y = 0.88xR² = 0.75
-1
-0.5
0-1 -0.5 0
Estim
ated
Ln(
N/N
0)
Measured Ln(N/N0)
Classical Chick-Watson Model
Delayed Chick-Watson Model
A. fumigatus type strain
A. versicolortap water
isolate
P. purpurogenumtap water isolate
y = 0.99xR² = 0.97
-5
-3
-1-6 -4 -2 0
Mea
sure
d …
Measured …
1 mg/L assay
4 mg/L assay
SI-Figure 5. Comparison of model predicted Ln(N/N0) against measured Ln(N/N0) for both delayed Chick-Watson model and classical Chick-Watson model, monochloramine disinfection
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Table SI-1. Comparison of estimated 3-log inactivation Ct values of the tested fungal strains to previously tested virus and bacteria, by free chlorine
Ct (mg min/L) References(Previously tested virus and bacteria, 3-log inactivation)Escherichia coli (pH 7.4) 0.032 ±0.009 (Helbling and
VanBriesen 2007)Staphylococcus epidermidis (pH 7.4) 0.221 ±0.08 (Helbling and
VanBriesen 2007)Klebsiella pneumoniae (22.5 ºC, pH 7.2) 13.06 ±0.91 (Goel and Bouwer
2004)Mycobacterium avium strain 5502 (23 ºC, pH 7) 51 ±10 (Taylor et al. 2000)Mycobacterium avium strain 1508 (23 ºC, pH 7) 164 ±28 (Taylor et al. 2000)
Mycobacterium aurum (pH 7.4) 42.9 ±2.71 (Helbling and VanBriesen 2007)
Legionella pneumophila (25 ºC, pH 7) 365.3 (Jacangelo 2002)
EPA required Ct for 3-log inactivation of Giardia (20 - 25 ºC, pH 7)
37 (Crittenden et al. 2005)
(Fungal strains in the current study, 3-log inactivation)Aspergillus fumigatus type strain (22.5 ºC, pH 7) 61.42 ±4.74Aspergillus fumigatus clinical strain (22.5 ºC, pH 7) 48.99 ±11.36Aspergillus versicolor (22.5 ºC, pH 7) 84.72 ±6.85Penicillium purpurogenum (22.5 ºC, pH 7) 194.7 ±21.6
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Table SI-2. Comparison of estimated 3-log inactivation Ct values of the tested fungal strains to previously tested virus and bacteria, by monochloramine
Ct (mg min/L)
References
(Previously tested virus and bacteria, 3-log inactivation)Adenovirus (19 ºC, pH 7) 1.24 (Jacangelo 2002)E. coli (25 - 35 ºC, pH 7.5 to 8.5) 10.3 ±1.67 (Jakubek et al. 2013)Corynebacterium glutamicum (25 - 35 ºC, pH 7.5 to 8.5)
16.84 ±1.18 (Jakubek et al. 2013)
Pseudomonas fluorescens (25 - 35 ºC, pH 7.5 to 8.5) 22.19 ±3.04 (Jakubek et al. 2013)Klebsiella pneumoniae (22.5 ºC, pH 7.2 to 7.5) 39.57 ±1.27 (Goel and Bouwer
2004)Mycobacterium avium strain 5502 (23 ºC, pH 7) 91 ±34 (Taylor et al. 2000)Mycobacterium avium strain 5002 (23 ºC, pH 7) 1710 ±814 (Taylor et al. 2000)Legionella pneumophila environmental strain 1 (25 - 35 ºC, pH 7.5 to 8.5)
16.14 ±3.07 (Jakubek et al. 2013)
Legionella pneumophila environmental strain 2 (25 - 35 ºC, pH 7.5 to 8.5)
64.88 ±19.07 (Jakubek et al. 2013)
EPA required Ct for 3-log inactivation of Giardia (20 ºC, pH 6-9)
1100 (Crittenden et al. 2005)
(Fungal strains in the current study, 3-log inactivation)Aspergillus fumigatus type strain (22.5 ºC, pH 8) 103.9 ±6.64Aspergillus fumigatus clinical strain (22.5 ºC, pH 8) 90.33 ±7.17Aspergillus versicolor (22.5 ºC, pH 8) 531.3 ±87.11Penicillium purpurogenum (22.5 ºC, pH 8) 153.2 ±9.07
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Reference:Crittenden, J.C., Trussell, R.R., Hand, D.W., Howe, K.J. and Tchobanoglous, G. (2005) MWH's Water Treatment: Principles and Design, Second Edition, pp. 1914-1917, John Wiley & Sons, Inc.Goel, S. and Bouwer, E.J. (2004) Factors influencing inactivation of Klebsiella pneumoniae by chlorine and chloramine. Water Research 38(2), 301-308.Helbling, D.E. and VanBriesen, J.M. (2007) Free chlorine demand and cell survival of microbial suspensions. Water Research 41(19), 4424-4434.Jacangelo, J.G. (2002) Inactivation of waterborne emerging pathogens by selected disinfectants, American Water Works Association.Jakubek, D., Guillaume, C., Binet, M., Leblon, G., DuBow, M. and Le Brun, M. (2013) Susceptibility of Legionella Strains to the Chlorinated Biocide, Monochloramine. Microbes and Environments 28(3), 336-345.Sivaganesan, M., Rice, E.W. and Mariñas, B.J. (2003) A Bayesian method of estimating kinetic parameters for the inactivation of Cryptosporidium parvum oocysts with chlorine dioxide and ozone. Water Research 37(18), 4533-4543.Taylor, R.H., Falkinham, J.O., Norton, C.D. and LeChevallier, M.W. (2000) Chlorine, chloramine, chlorine dioxide, and ozone susceptibility of Mycobacterium avium. Applied and Environmental Microbiology 66(4), 1702-1705.
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