PII: S0043-1354(01)00011-2

Wat. Res. Vol. 35, No. 13, pp. 31683178, 2001

# 2001 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
0043-1354/01/$ - see front matter

IN SITU MEASUREMENT AND STATISTICAL MODELLING

OF ESCHERICHIA COLI DECAY IN SMALL RIVERS
PASCAL BEAUDEAU1*, NICOLAS TOUSSET1, FRANCK BRUCHON2,
AMELIE LEFE`VRE1 and HUW D. TAYLOR3
1

Laboratoire dEtudes et dAnalyses de la Ville du Havre 70 quai Frissard, 76600 Le Havre, France;
Agence de lEau Seine-Normandie-Direction des Rivages Normands 21 rue de lHomme de Bois,
14600 Honeur, France and 3 The School of the Environment, Brighton University Cockcroft Building,
Lewes Road, Brighton BN2 4GJ, UK
2

(First received 17 July 2000; accepted in revised form 18 December 2000)

Abstract}Numerous studies have been carried out on the environmental factors associated with the decay
of faecal bacteria in open (fresh or marine) waters. The present study aimed at understanding the fate of
bacteria in small streams (ow 520 m3 s
1) for which there is a lack of knowledge. An original in situ
protocol was developed for measuring the die-o of Escherichia coli (E. coli) from wastewater treatment
plants. Based upon 80 values of the decay rst-order parameter (K or its inverse T90), collected from ve
rivers in Normandy (France), a median T90 of 10 h and a minimal T90 of 1.3 h were obtained. K was then
modelled as a linear function of variables made up from ow, water temperature and suspended
particulate matter (SPM). The set of signicant co-variables did not include light indicators. E. coli decay
is inversely related to the river ow and it becomes highly signicant below 0.3 m3 s
1. The positive eect
of small ows on die-o is increased by water temperature over 158C, whereas it could be reduced by SPM.
The major co-variable of the model ( p510
9 ) is an empiric composite variable integrating the eect of
ow and temperature that explains more than 40% of the variance of K. We interpreted this as an
expression of predation by benthic micro-grazers which could be the main cause of E. coli die-o in small
streams in temperate countries. # 2001 Elsevier Science Ltd. All rights reserved
Key words}Escherichia coli, rivers, decay model, ow, temperature, SPM

INTRODUCTION

Microbial contamination from faecal origin in

natural waters causes a sanitary risk through
consumption of drinking water, consumption of
shellsh and bathing. However, as it disappears
spontaneously, it is important to quantify its kinetics
of decay, for assessing the level of sewage treatment
required for the protection of sensitive areas.
Our study focused on bacterial indicators, since
European sanitary regulations are based on the
monitoring of this kind of indicator. Escherichia coli
(E. coli) was chosen because of its broad use as a
tracer for faecal contamination.
Usually, population dynamics for allochtone
bacteria are modelled by rst-order kinetics:
dNt=dt
kNt with k > 0, i.e. an exponential
decay of N with time t. This model is both simple and
ecient in some practical situations. Its single
parameter, i.e. the decay coecient k, is often
replaced by K k=2:3, which corresponds to the

*Author to whom all correspondence should be addressed.

Tel.:+33-141-796822; fax:+33-141-796768; e-mail:
p.beaudeau@invs.sante.fr

use of decimal logarithms for bacterial counts:

log Nt
log Nt0
Kt
t0 . K, usually expressed in h
1, and is thus the inverse of the period
of time T90h necessary for reducing the bacterial
population by 90%.
The decay coecient varies with the nature
bacteria and their physiological state, but also with
environmental factors. Numerous studies have focused on the environmental parameters inuencing
K, both in coastal waters and in some large rivers (see
Barcina et al., 1997 for a review), but no specic
work has been done on small streams. This study
aimed at evaluating the distribution of K in small
streams (ow within the range 0.120 m3 s
1), and at
pointing out the main environmental decay factors in
this type of environment. Use of laboratory experimental set-ups or microcosms set up in sea or river
locations was not acceptable here, as the goal was
to study the combined expression of hydrological
and ecological factors specic to small streams. This
experimental in situ approach required the development of original tools for measuring the decay
coecient.
This paper details both the eld experiment
used for data collection and the statistical model

3168

Escherichia coli decay in small rivers

expressing variations of K as a function of meaningful environmental variables.

STUDY RIVERS

The hydrographical surface network in Eastern

Normandy is poor (Fig. 1), as the underground
karstic network partially drains the chalk aquifer.
Five streams were selected for their typological
diversity and their conformity level to the ideal study
site as dened in the protocol section.
Table 1 shows the variety in sizes and features of
the selected rivers. Flows vary from 0.2 (Morelle) to
20 m3 s
1 (Risle). Current velocities are variable
between the rivers but also within each river, since
rapid current segments alternate with slower zones.
Because of numerous bends and obstacles lying in the
bed of the rivers, fast ows are typically turbulent
and thus ensure a good mixing of water throughout
the river section. Slower reaches however enable
SPM settling. The Risle area is distinguished by its
estuarine character. All the measurements in this area
were done around low tide in free-ow conditions.

3169

The watershed of the ve rivers studied are fairly

urbanised but mainly dedicated to animal husbandry.
The oxygenation level is always over 90% saturation.
Very low levels of organic pollutants are found in the
Morelle and the Ye`res, whereas moderate levels are
found in the Bethune and the Touques. By the
oxygenation it creates, the water movement generated by the tide seems to counteract the risk caused
by the high organic matter content found in the Risle.
Selection of river sections with an important source
of urban pollution upstream leads to high-average
concentrations of faecal bacteria.

PROTOCOL

The principle of the eld protocol is based on the

follow-up of a water mass transiting along the river
and on the measurement of bacterial ows
(and meaningful co-variables) at dierent points of
the transit sector. The practical application of the
principle is conditioned by certain characteristics of
the site: (i) presence of a regular and abundant pointsource of faecal pollution upstream, e.g. the outlet of

Fig. 1. Sites locating map.

Table 1. Hydrological characteristics of the studied sectors. Averages calculated on validated data (N 80)

Uspstream WWTP
Sector length (km)
River ow speed (m s
1)
Slope
Discharge (m3 s
1)
Maximal depth (m)
Width (m)
Turbidity (NTU)
SPM (mg L
1)
TOC (mg L
1)
O2 (% saturation)
[E.coli] (MPN/100 mL)

Morelle

Ye`res

Bethune

Touques

Risle

Beuzeville
3.8
0.21
1 : 130
0.22
0.20.5
1.63.2

Foucarmont
10.3
0.30
1 : 360
0.78
0.20.6
48

Neufchatel
7.6
0.18
1 : 550
1.4
0.81.7
511

Lisieux
18.0
0.41
1 : 700
5.4
1.12.5
1116

Pont-Audemer
8.0
0.52
1 : 10 000
18
1.72.6
1522

4.1
10.9
1.3
101
10 000

4.0
9.2
1.0
92
3 000

3.1
4.3
2.6
107
8 000

7.5
14.5
2.2
90
4 000

19.6
44.3
3.8
109
4 000

3170

Pascal Beaudeau et al.

an urban waste water treatment plant (WWTP). By

using this type of outlet, we take into account the
resistance acquired by the residual bacterial population during its transit through the WWTP (Dupray
and Derrien, 1995); (ii) absence of lateral sources of
faecal pollution along the studied area; (iii) a simple
hydrographic network, free from dead arms, divisions and losses, in order to ensure the conservation
of the tracked water mass. On the other hand, the
dilution of the contaminated main stream by some
unpolluted tributaries does not aect the decay
coecients assessment, because E. coli ows rather
than concentrations were measured; and (iv) a long
enough transit section, for the transit time to be
compatible with a measurable and signicant decay
of E. coli.
The sampling protocol controls the gaps between
the observed situation in the ve studied rivers and
the theoretical feature described above. The main gap
concerns the contamination of the studied water
mass during its transit by identied (e.g. contaminated tributaries, outlets of domestic waste water) or
unidentied sources (e.g. submerged outlets, diuse
pollution). In order to minimise this problem, it was
necessary to carefully select adapted sites but also to
choose favourable hydrological conditions. Temporary contamination generated by run-o or resuspension of contaminated sediments deposited in the river
bed were avoided by undertaking all the experiments
during the hydrological recession phase and in the
absence of precipitation.
Data acquisition
Assessing the decay coecient K D log F=Dt
requires measuring (i) the transit time Dt of a water
mass between two reference points, and (ii) at these
points and on passage times, the bacterial concentration (C) and the river ow (Q), the product of which
gives the bacterial ow F CQ. Dt and Q were
measured by tracer studies.
Tracer studies. The measurements of ow and
transit time were carried out by chemical tracer
studies. Two tracers were used according to the ow:
a brackish solution of 100300 kg of sodium chloride
(NaCl) for small ows (53 m3/s) or a solution of 1 kg
of lithium chloride (LiCl) for larger ows (>3 m3/s).
Concentrations of tracer after dilution in the river
ow were not toxic to fauna nor to E. coli. The
transit time was assessed using the passage of the
concentrations peak.
NaCl concentrations were estimated from in situ
conductivity measurement according to the Dienert
formula, knowing that the ratio of the two parameters is linear in the range of encountered
concentrations (Rodier, 1996). Measurements were
performed with a WTW LF196 conductimeter, with
a resolution of 1 mS cm
1, corresponding to 0.53 mg
NaCl L
1. Real-time sampling for evaluation of

bacterial ows was then possible when using NaCl

tracing.
Li was dosed in the laboratory by ame atomic
absorption spectrometry (VarianTM spectrometer,
model SpectrAA 300) which enabled a 1 mg L
1
detection threshold to be met. When LiCl was used,
bacterial samplings were performed one day after the
tracing.
Flow measurements required recovering the whole
mass of tracer, and the conversion of conductivity
into NaCl concentration. Applying the R mass
conservation law for the tracer mass Mtrac tr Q 
tCtrac t dt enabled the calculation of the ow Q,
supposed to be constant during the transit time
interval t. Discrete measurements collected with the
sampling lap
P time Dti lead to the approximation
Q # Mtrac = i Ci
C0 Dti , where Ci is the measured concentration and C0 the background level for
the tracer. The precision of Q assessment (about 10
20%) remained high in comparison to precision of
bacterial concentrations, and thus was not responsible for a signicant part of the uncertainty in the E.
coli ow estimate.

Sampling and analyses

Sampling. Sectors on the rivers were divided into

2 or 4 (Morelle and Ye`res) consecutive sections, on
which decay coecients were calculated. Each daytime experiment was repeated during the following
night. Night sampling was based on transit times
estimated during the day.
For one ow measurement, eight 500 mL-samples
(12 for the most upstream point) were realised with
1 min delay between each. Samples were transported
and stored at 48C. Two average samples (3 for the
upstream point) were prepared with four consecutive
point samples each. Analyses were performed less
than 24 h after sampling. Former experiments (not
related here) showed the absence of any shift in
numbers within this delay. As E. coli concentrations
are assumed to be log-normally distributed, geometric averages of the 2 or 3 analytical results were
used for ow calculation. Moreover, repeating the
analyses enabled us to assess the instant variability of
E. coli concentrations for each sampling point and so
to test the homogeneity of the measurements
involved in one ow calculation. We thus checked
the good cross mixing of the euent with the river
water and the steadiness of E. coli ow from the
WWTP in the upper end of the sector. In the case of
signicant dierences among measurements, the
experiment was invalidated.
During each experiment, lateral E. coli ows from
the tributaries were submitted to measurements
synchronised with the water mass passage in order
to correct the main ow for their inuence (see the
following section).

Escherichia coli decay in small rivers

E. coli concentration measurement. Concentrations in E. coli were most-probable-number (MPN)

estimates drawn from microplate method achieved
by Maul and Block (1983) and by Hernandez et al.
(1993) in its normalized E. coli version (ISO 9308.3).
This method is based upon the uorogenic ability of
E. coli grown in a A-1 broth modied by adding
4-methylumbelliferyl b-d-glucuronide.
Each well is lled with 200 mL of diluted sample
water. The 96 wells are split into four groups of 24
wells corresponding to dilutions of 1/2, 1/20, 1/200
and 1/2000, as recommended for surface fresh waters
(ISO 9308.3). This gives bacterial counts in a range
between 40 and 3.2  106 MPN/100 mL.
Hernandez et al. (1991a,b) showed that this
method was as sensitive as usual methods (membrane
ltration, pour plate count, and 3 or 5 tubes mostprobable-number test), but more specic than these.
Moreover, the method is more precise and reproducible than 3 and 5 tubes most-probable-number test,
because MPN estimates are drawn from 96 wells.

Physico-chemical
measurements. Temperature,
conductivity and turbidity were measured in the eld
(hand-carried turbidimeter HACHTM 2100P). SPM
and total organic carbon (TOC) were analysed in
the laboratory according to normalised methods
(EN 872:1996 and EN 1484:1997, respectively). The
discharge of the tributaries was recorded by a Silex
InternationalTM 2.100 current meter according to
ISO/R 748 recommendation. Particular attention was
focused on the estimation of light intensity irradiating bacteria. This estimation was made in three steps.
The gross light power was recorded in the open eld
during each bacterial tracking with an integrative

3171

pyranometer (CM3 type cell, radiation indicator type

CC20 Kipp & ZonenTM ). Then, the reduction of light
intensity by bank vegetation and the average light
intensity available for bacteria inside the water
column were estimated.
Control of lateral contamination. An iterative
algorithm was programmed for the correction of K
values from the inuence of lateral pollution sources.
As F1 and F2 were ows of E. coli measured at both
ends of the considered section at passing times t1 and
t2 , initial value
K0

log F1
log F2
t2
t1

corresponded to the slope of the linear segment

linking the points (log F1 ; t1 ) and (log F2 ; t2 ). The
parameter K0 (K value at step 0) and the average
tracer speed on the section were applied to the
J lateral discharges fj j : 1; J for estimating residual
ows at the downstream point (denoted f2; j0 at
step
P 0). Replacing F2 by corrected ow F2;0 F2

J f2; j0 in Eq. (1) then enabled the recalculation of
the decay coecient. This updated and more
accurate value K1 , is then used at step 1 to assess
again residual ows of lateral discharges f2;j1 , etc. The
procedure is performed as many times as needed to
converge towards the end-point value K of the decay
factor. If lateral contamination was properly taken
into account, the successive values of K did not vary
very much between contiguous sectors during one
experiment along one river sector. On the contrary,
an unidentied lateral contamination induced a shift
towards lower values of K downstream. Sections
where such a bias was observed were abandoned
(Fig. 2).

Fig. 2. Evolution of the E. coli ow in the river Morelle (July 67 1995). Flows values results from
lagrangian tracking of a marked water mass. Bacteria are discharged by an urban waste water treatment
plant located 1 km upstream from the head of the study sector. No signicant additional pollution occur in
the rst two sections, so that the decay coecient K is apportioned to slope of both corresponding
segments of the line. On the contrary, the upgoing trend of the line on the two last sections indicates
parasitic pollution inputs. These sections were afterwards excluded from the sampling.

3172

Pascal Beaudeau et al.

Statistical analysis
In the multifactorial modelling of decay coecient
Mancini (1978), Auer and Niehaus (1993), and
Wilkinson et al. (1995) express this inclusive coecient as the sum of the specic coecients for each
decay factor. Thus, we adjusted the observed decay
coecients by multiple linear regression to variables
corresponding to decay factors, using the leastsquare criterion. A stepwise procedure was carried
out for selecting signicant variables. Because there
were noticeable dierences in the degree of condence about K measurement, K values were
weighted according to their degree of condence,
i.e. by the transit time, before tting the model.
The variance of K could be regarded as the sum of
(i) the variance introduced by measurement process
(varm) and (ii) the variance of the decay itself
attributable to endogenous (bacterial physiology,
etc.) and exogenous factors (environment), as it
could be assumed that both sources of variance do
not interfere. Raw measurements used for the
calculation of K were bacterial concentrations,
stream ows and transit times. Errors made on times
and ow measurements are weak compared with the
error made on bacterial concentrations. Making
approximations that (i) ows and transit times are
perfectly known and (ii) ows do not vary throughout the section, yields:


D logQC
varm K varm
Dt
 Dt
2 varm D log C

Repeated measurements of E. coli concentrations

enabled us to assess the variance of instant log C
measurement at each sampling point, resulting from
both analysis and sampling (small scale heterogeneity
of water). Fischers test showed there was no
signicant dierence of log C variance attributable
to measurement between sampling points nor rivers.
So varm D log C could be taken as a constant
among the whole data set and varm K only
depended on the transit time as indicated in Eq. (2).
Thus, weighting K values by the associated transit
time resulted in equalising the measurement-related
contribution of each point to the total inertia of the
set of observed values of K, and prevented lowcondence points (likely to be thrown o the centre
because of measurement errors) from driving the
tting of the model.
The statistical validity of the linear model is based
on the compliance with hypotheses concerning the
distribution of rK residuals of K. The Kolmogorov
Smirnov test was employed to check if rK residuals
tted a gaussian distribution. The Fisher test enabled
us to estimate the constancy of rK variance for any
level of K. If both hypotheses were satised, the covariables selection procedure, based on Students Ttest, was a posteriori justied. Introduction of

quadratic terms of the co-variables into the model

was attempted in order to point out non-linearity.
To complete the statistical approach, the sensitivity
of the results (i) to mis-tted individuals or atypical
rivers (e.g. small-sized Morelle and estuarine Risle)
and (ii) to alternative tting options (uniform
weighting) was studied.
The statistical software SPSS/PC+TM was used for
statistical modelling.
RESULTS

Forty-four eld experiments were carried out

between June 95 and June 96, except during winter
(DecemberFebruary), each one including one tracer
study, one day-time and one night-time sampling.
This eld study resulted in a total of 80 validated
values of K. Lateral E. coli ows brought by
tributaries did not disrupt measurements, since only
9% of K values corresponded to cases for which
cumulated lateral ows were above 10% of main
ow. The observed concentrations of E. coli ranged
from 6  104 MPN/100 mL in the upstream sections
to 2  102 MPN/100 mL in the downstream sections
(Table 2). The distribution of K values (Fig. 3 and
Table 2) showed a median of 0.10 h
1 (T90 10 h)
and a maximum of 0.76 h
1 (T90 1:3 h), that
clearly indicated an overdispersion of the highest
values. K values from small-size rivers (Ye`res and
Morelle) were spread over a large range and included
all K values above 0.25 h
1 (T9054 h).
The overall estimate of the standard error on
log C averaged values used for ow assessment
(see
the section
Sampling and Analysis) was

q
varm log C 0:095. Large dierences in the

transit times were observed, with a 13 h range for

the Morelle, Ye`res and Risle sections, and a 38 h
range for the Bethune and Touques sections. In this
respect, condence of K (equation (2)) is better for
the Touques and the Bethune than for the other
rivers, which bears out the use of weighting before
tting the model.
The model with K as dependent variable
The only signicant co-variables gathered in the
linear model were the ow Q, the temperature T and
the SPM content, the observed distribution of which
are given in Table 2. Among the other variables
tested, neither the organic pollution level, nor the
light intensity, were integrated to the model.
Raw variables Q, T and SPM however needed
special design in order to work eciently for K
forecasting. We suggest an interpretation of
the transformed variables in the discussion.
The power function of the ow Q
0:50 , the
exponent of which is empirically adjusted, is the
main determinant of K (R2=0.36; p510
7 ).
No signicant correlation was observed between K
and the temperature, taken as a true variable.

Escherichia coli decay in small rivers

3173

Fig. 3. Count histograms of observed values of the decay coecient K.

Table 2. Distribution of E. coli concentration and indicators used in models 1 and 2 (N 80)

Minimum
Perc. 5%
Median
Perc. 95%
Maximum

[E. coli] (MPN/100 mL)

K (h
1)

Q (m3 s
1)

T (8C)

SPMr

2.4  102
6.9  102
5.2  103
4.0  104
5.5  104

0.004
0.020
0.106
0.415
0.744

0.16
0.16
1.05
19
23

6.9
7.1
11.9
15.5
18.3

0.12
0.36
1.02
1.70
2.26

However, temperature signicantly acted on K

through a composite variable made up both
from ow and temperature. The optimised shape
of the composite variable, i.e. maximising the
correlation with K, resulted from the product of
the former Q variable by the temperature term used
by Menon (1993) for modelling the level of protozoa
grazing; in the river Seine (France): QT Q
0:50 exp

25
T2 =202 with T expressed in 8C and Q in
Ls
1. The correlation between QT and K ( p510
9 )
signicantly exceeded that obtained without
the temperature term. A search algorithm, performed in order to determine the 3 QT parameters
(i.e. the 2 T parameters and the Q exponent) values
optimising the correlation with K, showed that the
values selected by Menon (1993) for the two
temperature parameters were almost optimal
(  5%).
QT eect on K seemed inferior at night than during
day-time, as shown by breaking down QT into 2
variables according to
*

QTd=QT for a day-time measurement and

QTd=0 elsewhere

QTn=QT for a night-time measurement and

QTn=0 elsewhere

The variable SPMr, ratio between SPM content

observed in the section during the experiment and the
average calculated on the whole data set available for
that section, also contributed signicantly ( p50:05)
to the variation of K.
Finally, we suggested the following equation where
rK stands for the model residual:
Model 1: N 78, R2 0:45
K = 0.069 + 5.4QTd+4.0QTn
0.050 SPMr+rK
[0.02; 0.12] [3.9; 6.9]

[2.5; 5.5]

[
0.09;
0.01]

CI(95%)

We pointed out 2 outlying points (p510
5 ),

corresponding to the maximum values of K
(T90 1:3 and 1.4 h), and eliminated them in the
nal formulation of the model.
No signicant correlation was observed between
the co-variables QT and SPMr.
Simulations of model 1 (Fig. 4) showed that values
for the variable QT , and consequent expected values
of K, evolved moderately when the ow remained

3174

Pascal Beaudeau et al.

Fig. 4. Predictions Ke of the decay coecient K given by models 1 (top) and 2 (bottom) for SPMr=1 and
by day-time.

higher than 0.3 m3 s
1 and temperatures below 158C,

then increased rapidly towards small ows and high
temperatures.
The model was robust to all the variations
operated on the weighting system or on the data
set used for its adjustment, i.e. the major variables
and the strength of association were conserved
(Table 3). Introduction of quadratic terms of
signicant co-variables did not improve the quality
of the tting, which conrms the linearity of the
relation between K and its expected values Ke
provided by model 1, as suggested by Fig. 5. The
KolmogorovSmirnov test showed that the distribution of the model residual rK did not dier
signicantly from gaussian, even for a rejection
threshold of 10%. To test the constancy of the
residual rK distribution along the range of Ke values,
we divided the variation domain of Ke into 3

equirepresented classes (Table 4). We noticed a very

signicant increase of the variance as a function of
Ke: between the rst and the last third of the domain,
the ratio of variances for rK was F 10:1 (p510
6 ).
Change in variance thus prevented conclusions on
the models validity by using classical tests. However,
Spearman distribution-free test of ranks conrmed
the very signicant monotonous dependence between
K and Ke (p510
7 ). Even if not statistically fully
validated, model 1 clearly keeps a real descriptive
potential.
The model with log-K as dependent variable
In model 1, the approximate proportionality
between the standard deviation of rK and the level
of previsions Ke suggested the log-transformation of
KlK log K, which yields model 2

Escherichia coli decay in small rivers

3175

Table 3. Sensitivity study of the linear model expressing K (h
1) as a function of co-variablesa
Data weighting

N
R2
Variables:
QTd
QTn
SPMr
Constant
a

Data removal

8i, wi Dti

8i, wi 1

78
0.45

78
0.44

5.35
(10
9)
4.00
(10
7)

0.050
(0.05)
0.069
(0.01)

4.60
(10
9)
2.81
(10
3)

0.112
(NS)

Risle

Morelle

KI j pKi 510
3

58
0.33

76
0.52

65
0.47

linear coecients and (P-values)

5.65
5.21
(10
9)
(10
5)
4.25
2.32
(10
6)
(0.05)

0.054

0.057
(0.05)
(0.05)
0.065
0.095
(0.05)
(10
3)

5.47
(10
9)
3.55
(10
5)

0.055
(0.05)

0.062
(NS)

Column 1 corresponds to the reference model 1. In column 2, individuals are given the same weight (8i, wi 1). Columns 35 correspond to
the removal of specic data categories: rivers showing extreme ows (estuarine Risle and Morelle); values with a probability lower than
10
3.

Fig. 5. Observed values of K vs. expected values Ke provided by models 1 and 2 and the regression lines
(N 78 and 79, respectively).

Model 2: N 79, R2 0:38

Table 4. Evolution of the variance of the residual rK of model 1 as a
function of the expected decay coecient Ke
Domain of Ke

Variance of rK

Ke
0:078
0:0785Ke
0:147
Ke > 0:147

26
26
26

1.4  10
3
4.0  10
3
14.2  10
3

lK=
1.12 + 18.3QTd+13.8QTn
0.29SPMr+rlK
[
1.34;
0.90] [12.1; 24.5]

[7.4; 20.2]

[
0.47;
0.10]

CI(95%)

Model 2 used the same independent variables as

model 1. The role of the ow remained very
signicant (p510
4 ) whereas the role of the tem-

3176

Pascal Beaudeau et al.

Table 5. Part of the variance of K (model 1) or lK (model 2)

explained by the model according to the selected co-variables

Full model
Substitution of QTd QTn by . . .
QT
Q
0:50
Withdrawal of SPMr

Model 1

Model 2

(N 78)

(N 79)

45.4%

37.5%

42.8%
41.0%
41.5%

35.8%
33.5%
29.7%

Table 6. Evolution of the variance of the residual rlK of model 2 as a

function of the expected log-transformed decay coecient lKe
Domain of lKe

Variance of rlK

lKe5
1:21

1.21
lKe5
0.89
lKe

0:89

26
27
26

0.095
0.123
0.091

perature and SPM markedly reinforced (Table 5).

The signicance of SPMr increased to p50:005.
The log transformation made a new outlier appear
within the domain of low values of K, whereas the
two exceptionally high values of K (T90
1:4 h)
were re-integrated into the active data set. The global
correlation coecient was weakened relatively to
model 1, but the distribution of rlK residues fully
satised the conditions of use for classical tests,
particularly the constancy of the variance (Table 6
and Fig. 5).
Model 2 forecast a much higher gradient of K than
model 1 in the domain T > 158C and Q50:3 m3 s
1
(Fig. 4), thus following the exponential relationship
between K and the two independent variables:
K a expQbT exp
SPMr. This re-writing of
model 2 as a product also suggested the existence
of a synergy between QT and SPMr.
As model 1, model 2 is robust in the way it always
included the same pool of candidate variables with
steady-parameter values, whatever the variations
operated on the data set or on the weighting options.

DISCUSSION

Various decay coecients values were observed in

the studied river sectors, which may reach high values
in small streams. Reported maximum K values
measured for E. coli in freshwaters vary between
0.03 and 0.06 h
1 (30 h5T90515 h) according to
Barcina et al. (1986), Auer and Niehaus (1993),
Menon (1993), Mezrioui et al. (1995), but maximum
values as far from this interval as 0.001
(T90 40 days) and 0.25 (T90 4 h) were also
reported by Davies and Evison (1991) and by Evison
(1989), respectively. Reported maximum K values
usually remain much lower than those measured in

this study, except for the biggest rivers studied here,

i.e. Risle and Touques.
Precautions must be taken in the analysis of this
dierence. Analytical techniques used in this work,
based on bacterial ability to grow on culture media,
may lead to a noticeable overestimation of bacterial
decay due to loss of culturability (Kapuscinski and
Mitchell, 1981; Barcina et al., 1989). However, the
dependence between the result and the numeration
technique used (cultivation based technique vs. direct
counting) mainly concerns abiotic factors, such as
light, which cause stressed states between culturability and death. Decay factors suggested from this
study (see below) were only factors causing the direct
destruction of the cell, i.e. predation, or matrix
change, i.e. sedimentation. Thus, no major interaction is expected between the chosen analytical
method and the values of K obtained.
Ecological factors specic to small streams could
more likely explain the fast-decay kinetics observed
in this study.
Despite a specic eort made in the assessment of
light available for bacteria inside the water column,
its role in E. coli die-o did not emerge from this
study. The positive association between K and the
bacterial exposure to light showed a poor signicance
(p50:05), but partial correlation dropped to a nonsignicant level, if the main factor (resulting in
variables QTd and QTn ) had been properly controlled.
In contrast, as much as half of the variance of K
was explained by the ow, and a specic design was
needed to maximise its correlation with K. The
question of the underlying process is thus asked.
Assuming a perfect vertical mixing of water due to
turbulent ow, the probability of contact between
bacteria and bed surface varies commensurately with
the exchange surface between the water mass and the
river bed (specic contact surface). This probability is
inversely proportional to the wetted perimeter. The
power function of the ow Q
0:50 , used in the
variable QT , was empirically linked to the river size,
expressed by averaged width (R2 0:89), depth or
wetted perimeter. Q
0:50 is thus a reliable indicator
of the specic contact surface, more regular and
generally better-known than any size measurement.
The contact between bacteria and bed surface,
expressed by Q
0:50 , could result in capture by
benthic biolm and/or predation. McCambridge
and McMeekin (1980) showed the eect of temperature on the activity of predators of E. coli. As the
multiplication of Q
0:50 by a temperature term
expressing protozoal activity level in local fresh
waters (Menon, 1993) and resulting in QT , improved
the correlation with K ( p510
9 vs. p510
7 ), we
assumed that underlying phenomena involved in that
correlation were not purely physical but included a
biological component. The additional increase in the
correlation between K and QT obtained through
night/day distinction may also be interpreted as a

Escherichia coli decay in small rivers

biotic inuence, e.g. an increase in E. coli vulnerability to predation at light (Mitchell and Chamberlin, 1978; Fujioka et al., 1981; McCambridge and
McMeekin, 1981) and/or a lower predators activity
during night than day. Finally, adhesion to biolm
and benthic predation, expressed by the couple of
variables (QTd ; QTn ), could be the main determinant
of E. coli decay in small streams.
Auer and Niehaus (1993), Wilkinson et al. (1995)
consider co-sedimentation, i.e. the joint deposition of
particles and bacteria, as the only way for bacteria to
sediment in rivers. The variable SPMr was computed
as the ratio between SPM content observed in the
section during the experiment and its average
calculated during the whole study for that section.
For steady sources of SPM, SPMr could indicate the
depositing potential of ow (the higher SPMr, the
lower the depositing potential), as did the dierence
between actual ow and average ow in Wilkinson
et al.s study (1995). Co-sedimentation may thus
occur in slowering reaches of studied rivers. The logtransformation of K (model 2) reinforced very
perceptibly the role of SPM (Table 5) and suggested
a synergy between co-sedimentation and capture by
benthos.
Both models 1 and 2 met conceptual predation
models. First-order decay kinetics, with a coecient
apportioned (model 1) or quasi-apportioned (model
2) to the predators grazing level QT can be regarded
as a simplication (i.e. term expressing growing
removed) of LotkaVolterra preypredator models
(Wangersky, 1978).
The design and our interpretation of meaningful
variables for K assessment are summarized in Table
7. If benthic predation emerges as the more likely
cause of E. coli decay in small rivers, its feeding may
dier according to the ow conditions. Within fast
current reaches, bacteria would meet benthic predators only by the mixing ensured by turbulences,
whereas in slowered ow reaches, co-sedimentation
may partly supply benthic predators with bacteria
from the water column.
Though the main role of predation in the decay of
allochtone bacteria in surface waters is admitted by

3177

many authors as McCambridge and McMeekin

(1980, 1981), Servais et al. (1985), Barcina et al.
(1991), Menon (1993), Menon et al. (1996), Mezrioui
et al. (1995), Wilkinson et al. (1995), the specic role
of benthos has not been signicantly reported.
Menon (1993) only mentioned that stalked protozoa
of the genera Vorticella might contribute to half of
the predation of allochtone bacteria in the Seine
downstream of Paris. Lack of clear references
to benthos may have two reasons, one linked to
the choice of sites, the other to the choice of the
experimental protocol. The important size of the
rivers studied by McCambridge and McMeekin
(1981), Menon (1993), Wilkinson et al. (1995)
determines a predation dominated by plankton as
the role of benthos weakens very remarkably when
the ow exceeds 10 m3 s
1 (according to our results).
Because of its strong link to the ow, evidencing this
factor through statistical modelling of K requires
studying a large range of river sizes. Moreover,
widely used experimental protocols based on immersed diusion chambers, which physically prevent
benthic predation, seem to be fundamentally unsuitable for understanding faecal bacterial decay in
small-size rivers.

CONCLUSIONS

The main and highly signicant variable for a

prediction of decay coecient of E. coli in small
streams was the ow, designed in order to indicate the
intensity of the contact between water and river bed.
Including temperature, night/day distinction and SPM
improved the performances of models, especially if K
is previously log-transformed. The strength of the
associations, their robustness and their interpretability
as the expression of benthic predation supported the
consistency of the suggested models. However a
biological approach has to be completed in order to
test the hypothesis of benthic predation.
Models 1 and 2 kept close to one another in the
way they were based upon the same raw co-variables
and only diered in the shape of the relationship

Table 7. Co-variables of the decay coecient of E. coli in small streams

Gross variable

Charge Q (L s
1)

SPM concentration (mg L
1)

Temperature T (8C)
2

Variable shaped for

linear modelling

f Q Q
0:50

f T exp

25
T
202

!
SPMr

SPM a
SPM

Solar irradiation
Ecient light powerb

QT f Q f T
Signicance

NS

NS

Sedimentation

Light inactivation

#
Interpretation
a
b
c

Water/bed contact
Pelagic predation
Benthic predation

SPM is the average concentration observed during the study at the considered sampling point.
Estimation of the average light power received by bacteria.
NS: non signicant. *p50.05; #p510
5.

3178

Pascal Beaudeau et al.

between the decay coecient and these co-variables.

Statisticians would prefer model 2 which satised the
conditions for carrying out classical tests. Moreover,
model 2 foresaw correctly the highest values of K
which are outliers for model 1. In practice, the
assessment of SPMr requires (i) an historical record
of SPM measurements and (ii) standardised procedures for selecting data on which average SPM over
hydrological recessions may be computed. By
default, SPMr could be xed to 1, but that makes
more dicult the use of model 2 because of its
particular sensitivity to SPMr.
Insofar as we limited the study to E. coli and we
purposely avoided winter conditions, attention must
be paid to the conditions of use of the proposed
models. These models can help forecast the abatement of E. coli concentration in small rivers during
summer, and guide decisions in waste water treatment towards the protection of bathing areas.
The study reported herein contributes to the
knowledge of the survival patterns of E. coli within
temperate fresh water climates. It is, however,
appreciated by the authors that our conclusions
may lead to an over-extension of public health
protection level if they are improperly extended to
pathogens such as viruses, protozoa and even some
bacteria, (i) which better resist adverse conditions
than E. coli and (ii) the decay of which may be
controlled by specic environmental factors.
Our models are also of poor use for the protection
of seashell raising sectors, whose most serious
problems usually occur in winter. In winter, frequent
rainfalls induce huge variations in E. coli inputs
whereas spates cause massive resuspensions of
bacterial populations formerly settled with particulate matter. Consequently, analytical models based
on dierential equations describing not only predation but also transfers of water, sediments and faecal
bacteria, are required for a reliable forecast in these
specic conditions.
Acknowledgements}We thank the Direction des Rivages
Normands of the Seine-Normandie Water Agency who
nanced this work. We are also thankful to Paul Ferlin for
his contribution to selecting the sites, Jean-Francois
Lhuissier for his help in light measuring, Jean-Francois
Hernandez and Axel Hartke for their advice on writing the
paper.
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