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326

Prevalence of Escherichia coli O157:H7 and


Salmonella spp. in surface waters of southern
Alberta and its relation to manure sources
J.Y.M. Johnson, J.E. Thomas, T.A. Graham, I. Townshend, J. Byrne, L.B. Selinger,
and V.P.J. Gannon

Abstract: The Oldman River watershed in southern Alberta, Canada, is an extensively irrigated region in which intensive
agricultural practices have flourished. Concern over water quality in the basin has been expressed because of high levels
of enteric disease indigenous to the region. To address these concerns, we conducted a 2-year study to estimate the
prevalence of Escherichia coli O157:H7 and Salmonella spp. in surface water within the basin. This study is the first
of its kind to identify E. coli O157:H7 repeatedly in surface water collected from a Canadian watershed. Prevalence of
E. coli O157:H7 and Salmonella spp. in water samples was 0.9% (n = 1483) and 6.2% (n = 1429), respectively. While
data examined at a regional level show a relationship between high livestock density and high pathogen levels in southern
Alberta, statistical analysis of point source data indicates that predicted manure output from bovine, swine, and poultry
feeding operations was not directly associated with either Salmonella spp. or E. coli O157:H7 prevalence. However,
geography and weather variables, which are likely to influence bacterial runoff, were not considered in this model. We
also postulate that variations in time, amount, and frequency of manure application onto agricultural lands may have
influenced levels of surface-water contamination with these bacterial pathogens.
Key words: water, Salmonella, E. coli O157:H7, manure output, agriculture.
Rsum : Le bassin versant de la rivire Oldman, dans le sud de lAlberta, Canada, est une rgion fortement irrigue
o des pratiques dagriculture intensive se sont dveloppes. Des inquitude quant la qualit de leau dans le bassin
ont t mises en raison des taux levs de maladies entriques qui svissent dans la rgion. Pour rpondre ces inquitudes,
une tude dune dure de deux ans a t accomplie afin dvaluer la prvalence de Escherichia coli O157:H7 et de
Salmonella spp. dans les eaux de surface du bassin. Cette tude est la premire de ce type identifier du E. coli
O157:H7 rptition dans de leau de surface dun bassin versant canadien. La prvalence de E. coli O157:H7 et de
Salmonella spp. dans les chantillons deau tait de 0,9 % (n = 1483) et de 6,2 % (n = 1429), respectivement. Bien
que les donnes au niveau rgional qui ont t examines rvlent un rapport entre la densit leve du btail et les niveaux
levs de pathognes dans le sud de lAlberta, lanalyse statistique des donnes des sources ponctuelles indique que la
production projete de fumier provenant de lalimentation dlevages bovins, porcins ou de volaille ntait pas directe-
ment associe la prvalence de Salmonella spp. ou de E. coli O157:H7. Toutefois, les variables gographiques et cli-
matiques, qui pourraient influencer le ruissellement de bactries, nont pas t retenues dans ce modle. Nous postulons
galement que les variations de temps, de quantit et de frquence dpandage de fumier sur les terres agricoles ont pu avoir
un effet sur la contamination de leau de surface par ces bactries pathognes.

Mots cls : eau, Salmonella, E. coli O157:H7, production de fumier, agriculture.


[Traduit par la Rdaction] Johnson et al. 335

Introduction waters used by rural and urban centres. Approximately 10%


of the Oldman River watershed is irrigated agricultural land.
Southern Alberta is a semi-arid region in which the Irrigation waters are used for the production of feed needed
Oldman River drainage basin is the principal source of water to maintain intensive livestock operations, for livestock wa-
for agriculture, industry, and both drinking and recreational tering, as well as for recreation and residential purposes.

Received 23 January 2003. Revision received 9 June 2003. Accepted 10 June 2003. Published on the NRC Research Press Web site
at http://cjm.nrc.ca on 10 July 2003.
J.Y.M. Johnson,1 J.E. Thomas,2 and L.B. Selinger. Department of Biological Sciences, University of Lethbridge, Lethbridge, AB
T1K 3M4, Canada.
T.A. Graham and V.P.J. Gannon. Health Canada, P.O. Box 640, Lethbridge, AB T1J 3Z4, Canada.
I. Townshend and J. Byrne. Department of Geography, University of Lethbridge, Lethbridge, AB T1K 3M4, Canada.
1
Present address: Department of Public Health Sciences, University of Alberta, Edmonton, AB T6G 2G3, Canada.
2
Corresponding author (e-mail: thomas@uleth.ca).

Can. J. Microbiol. 49: 326335 (2003) doi: 10.1139/W03-046 2003 NRC Canada

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Johnson et al. 327

Since 1991, the livestock industry in Alberta has ex- River; and west of the Little Bow River (Figs. 1, 2, and 3),
panded significantly. Between 1991 and 2001, the number of so that water samples were representative of the region. Al-
cattle and calves in Alberta increased from approximately though livestock feeding facilities were situated throughout
4.7 to 6.6 million (Alberta Agriculture 1991; Alberta Agri- the region with a nonhomogenous distribution, sampling
culture Food and Rural Development 1996; Statistics Can- sites were selected to reflect surface-water flow and not dis-
ada Online 2002). From 1996 to 2001, the total number of tribution of livestock density. In 1999, 522 samples were
pigs on Alberta farms grew from 1.7 million to more than collected from 69 sites. During the following year, 84 sites
2.0 million and the total number of poultry from 10.6 to were sampled and 961 samples were collected, including 38
13.3 million (Statistics Canada Online 2002). Much of this samples from the City of Lethbridge storm drain system and
activity has been centred in southern Alberta, a region that six samples from six municipal sewage treatment plants
also has one of the highest levels of enteric disease in the within the Oldman River basin. Twenty-three sites were
province (Khakhria et al. 1996). sampled regularly in both years. Sampling sites were se-
Waterborne outbreaks of enteric disease have resulted either lected to assess surface water at locations of interest
when public drinking water supplies were not adequately (nonrandomized sampling).
treated after contamination with surface water (Barwick et
al. 2000; Jones and Roworth 1996; Kondro 2000; Swerdlow Sample collection
et al. 1992) or when surface waters contaminated with en- Samples were taken either upstream from foothold or
teric pathogens have been used for recreational purposes from water at approximately 30 cm below the surface, using
(Keene et al. 1994; Kramer et al. 1996). While protozoa, a plastic container. Samples were collected in sterile, plastic
such as Giardia and Cryptosporidium (Roach et al. 1993; bottles; placed on ice; and transported to the laboratory for
Wallis et al. 1996), have been the focus of much concern in testing.
water contamination, enteric bacteria, such as Escherichia
coli O157:H7 and Salmonella spp., which are leading causes Detection of E. coli O157:H7
of gastroenteritis in Canada and the United States (Baxter Water samples were dispensed in 90-mL volumes into
and Morton 1987; Kramer et al. 1996; Levy et al. 1998), can clean, sterile culture bottles. To each of these bottles, we
also be a significant source of water contamination. From added a 10-mL volume of 10 buffered peptone water
1993 to 1998, water contaminated by E. coli O157:H7 (BPW) (Becton Dickinson, Sparks, Md., U.S.A.) and incu-
resulted in 454 cases of illness in the United States, while bated the bottles at 37C for approximately 6 h. At this
water contaminated with Salmonella species, such as Salmonella point, 1 mL was withdrawn, added to 9 mL of trypticase soy
typhimurium, infected 625 individuals and caused seven deaths. broth supplemented with 20 g/mL novobiocin (Difco, De-
Regions with high cattle density are prone to enteric dis- troit, Mich., U.S.A.), and held at 42C for 6 2 h.
ease (Johnson et al. 1999). Michel et al. (1999) performed Immuno-magnetic separation using magnetic beads coated
epidemiological studies that showed the incidence of human with E. coli O157 antibody was carried out, as described by
infection with verotoxin-producing E. coli (VTEC) in On- the manufacturer (Dynal, Oslo, Norway). After the last wash
tario between 1990 and 1995 was related to cattle density. In step, the anti-E. coli O157 magnetic beads were resuspended
Canada, Alberta has the greatest population of cattle and one in 50 L of wash buffer and streaked onto sorbitol MacConkey
of the highest levels of gastroenteritis resulting from infec- agar containing cefixime (ctSMAC) (0.05 mg/L) and tellurite
tion by Salmonella spp. and E. coli O157:H7 (Khakhria et (2.5 mg/L) (Sanderson et al. 1995). The ctSMAC plates
al. 1996). were incubated for 24 h at 42C and translucent colonies
In this study, we examined the prevalence of E. coli O157:H7 tested by slide agglutination, using E. coli O157-specific an-
and Salmonella spp. in surface waters within the Oldman tibody (Difco). Positive bacteria were inoculated into 5 mL
River basin in southern Alberta and explored relationships brain heart infusion broth (Difco) and grown at 37C over-
between the prevalence of these pathogens and both animal night. Each of these bacterial cultures was tested by PCR us-
agriculture and human population within the region. ing oligonucleotide primers that detect vt1, vt2, fliC, and
eaeAO157 genes (Gannon et al. 1997). Putative E. coli O157:H7
Materials and methods cultures were sent to the Health Canada Verocytotoxigenic
E. coli Reference Laboratory in Guelph, Ontario, for inde-
Study sites pendent confirmation.
The Oldman River basin is a semi-arid zone located in
southern Alberta, Canada. Total precipitation in Lethbridge, Detection of Salmonella spp.
the principal urban centre (1999 population 68 712), was Water samples cultured in BPW, from the pre-enrichment
231.6 mm from May to October 1999 and 130.5 mm from medium described above, were incubated for 1624 h at
May to October 2000 (information from the Agriculture and 37C. The pre-enriched culture was inoculated into two sep-
Agri-Food Canada Lethbridge Research Centre). Water sam- arate enrichment broths, Rappaport Vassiliadis (RV) broth
ples were collected from surface-water drainage from irri- and tetrathionate brilliant green (TTBG) broth. For the RV
gated agricultural regions, the Little Bow River, the Oldman broth, we inoculated 9.9 mL of the broth with 100 L of the
River, sewage treatment plants, and the City of Lethbridge BPW culture. For the TTBG broth, we inoculated a 9.0-mL
storm drains. Sampling sites were located east from the volume with 1.0 mL of the BPW culture. Both of the inocu-
Rocky Mountains, along the Oldman River; north, along the lated enrichment broths were incubated at 42C for 24 h.
Little Bow River and at inflow and outflow locations of the Following enrichment, both the RV and the TTBG broth cul-
Lethbridge Northern Irrigation District; north of the Oldman tures were plated onto two different selective culture plates,

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328 Can. J. Microbiol. Vol. 49, 2003

Fig. 1. Prevalence of Escherichia coli O157:H7 in surface water and the animal manure output (AMU) in southern Alberta from 1999
to 2000; Universal Transverse Mercator (UTM) coordinates Zone 11, Datum, North American Datum (NAD) 27.

modified semi-solid Rappaport Vassiliadis (MSRV) agar (Difco). Putative Salmonella-positive colonies were further
(DeSmedt and Bolderdijk 1987) and brilliant green sulpha tested using standard (Christensens) urea agar, triple sugar
(BGS) agar (Becton Dickinson, Mississauga, Ont., Canada). iron agar, lysine iron agar slants (DAoust and Purvis 1998),
For inoculation onto MSRV plates, a 20-L aliquot of each and sulphide indole motility (Difco) agar (Carter 1979).
culture was dispensed onto the outside edge of each individ- Those colonies exhibiting biochemical responses typical of
ual plate. The plates were incubated at 42C for 24 h. Motile Salmonella spp. were further tested using a PCR specific for
bacteria producing halos greater than 20 mm in diameter in the detection of the Salmonella invA gene (Chiu and Ou
the MSRV medium were subcultured onto MacConkey 1996). Salmonella culture isolates were confirmed at the
(Difco) agar by taking an inoculation loop of cells from the Health Canada Salmonella Reference Laboratory in Guelph,
leading edge of the halo and streaking onto MacConkey Ontario.
plates. Following inoculation, the MacConkey plates were
then incubated at 37C for 1624 h. For testing on the BGS Prevalence
agar plates, both the RV and TTBG broths were gently The prevalence of both E. coli O157:H7 and Salmonella
mixed, and an inoculation loop of each of the cultures was spp. in water samples was calculated as
plated onto BGS agar plates. The BGS agar plates were then
incubated at 37C for 1624 h. Following incubation, three [1] (No. samples from which pathogens were detected
red colonies from each of the BGS plates were plated onto total no. samples tested) 100
MacConkey agar plates. These MacConkey plates were then
grown at 37C for 1624 h. For both the MSRV and BGS Mapping
agar plating methods, suspect translucent colonies were Prairie Farm Rehabilitation Administration (PFRA),
picked from each of the MacConkey plates and tested for Lethbridge, provided data of mapped manure output from
agglutination, using Salmonella O antiserum Poly A-I & Vi confined animal agriculture operations in southern Alberta.

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Johnson et al. 329

Fig. 2. Prevalence of Salmonella spp. in surface water and the animal manure output (AMU) in southern Alberta from 1999 to 2000;
Universal Transverse Mercator (UTM) coordinates Zone 11, Datum, North American Datum (NAD) 27.

The numbers of animals for each confined livestock feeding swine operations included farrow-to-finish, weaning, and
operation in the region were estimated from information feeder hog farms; poultry operations included broiler, layer,
provided in field surveys, air surveillance photos, or expan- turkey, and unknown poultry farms. Sampling sites from
sion permits. Animal manure units (AMU) were estimated which more than two samples were collected and for which
as the product of the number of animals by type for each lo- there were bovine, swine, and poultry operations within
cation, the annual excreted nitrogen mass for animal types 15 km were used for AMUpathogen prevalence analysis.
(Natural Resources Conservation Service 1999), and the in- AMU estimates at these sampling sites were calculated as
verse of the mass of nitrogen required to fertilize 1 acre of the mean of triplicate measurements. To classify sampling
corn for 1 year in Alberta, 73 kg/year (PFRA). To obtain sites outside the City of Lethbridge limits and provincial
AMU estimates that correspond to water sampling sites, a park boundaries by livestock agriculture activity, we made a
150-m2 grid of AMU values over the study area was interpo- 5-km2 grid of mean AMU density over the study area. The
lated from the AMU point data for confined livestock opera- AMU density of all grid cells were divided into four equal
tions (ArcView 3.0, ESRI, Redland, Fla., U.S.A.). intervals and classified as low, medium, high, and very high
Interpolated values for each cell of the surface were calcu- AMU density.
lated as the sum of the products of the AMU operations
within a 15-km radius and the inverse of the distance to Statistical methods
those operations squared. In this study, we assume that The apparent prevalence of E. coli O157:H7 and Salmo-
transport of manure beyond a 15-km radius of an animal nella spp. associated with sampling sites was estimated as
rearing facility would be prohibitively expensive and, conse- the percentage of pathogen-positive samples of all samples
quently, unlikely. AMU surfaces were generated for bovine, tested for both pathogens. The change in prevalence among
poultry, swine, and combined animal feeding operations. sampling sites tested in both years was assessed using a
Bovine operations included confined beef and dairy farms; paired sample t test. The association between the prevalence

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330 Can. J. Microbiol. Vol. 49, 2003

Fig. 3. Prevalence of Salmonella spp. and Escherichia coli O157:H7 in surface water in southern Alberta and bovine manure output
(AMU) surface estimated using inverse distance weighting from confined beef and dairy feeding operations; Universal Transverse
Mercator (UTM) coordinates Zone 11, Datum, North American Datum (NAD) 27.

of Salmonella spp. and E. coli O157:H7 and the AMU of [3] logit(ECOLI) logit(ECOLITEST)
sampling sites was assessed using generalized linear models,
where inputs included AMU output from confined bovine, = + 1 BOVAMU + 2 SWNAMU
swine, and poultry livestock operations. The prevalence of
Salmonella spp. among sampling sites followed a Poisson + 3 PLTAMU
distribution; thus, a Poisson regression model was used
(SAS, SAS Institute Inc., Cary, N.C., U.S.A.). The model where ECOLI is the number of samples from a site positive
was for E. coli O157:H7, ECOLITEST is the number of samples
tested from that site, and other variables are as defined pre-
[2] ln(SALM) ln(SALMTEST) = + 1 BOVAMU viously. Type III likelihood ratio statistics were used to as-
+ 2 SWNAMU + 3 PLTAMU sess the strength of association for each animal type, and
residual deviance statistics were used to assess good-
where SALM is the number of samples in which Salmonella ness-of-fit. Logistic regression models were developed to de-
spp. was detected from a given sampling site; SALMTEST scribe the relationship between pathogen presence in water
is the number of samples tested from that site; is the inter- samples and AMU associated with the site from which the
cept; BOVAMU, SWNAMU, and PLTAMU are independent samples were collected, assuming the AMU at sites re-
variables; and 1, 2, and 3 are slopes (parameter estimates mained stable over the period of time in which samples were
for bovine, swine, and poultry AMU, respectively). The collected (SPSS 10.0.5, SPSS Inc., Chicago, Ill., U.S.A.).
prevalence of E. coli O157:H7 followed a binomial distribu- Model fit was assessed using the Hosmer and Lemeshow
tion, thus, the following logit regression model was used: goodness-of-fit test.

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Johnson et al. 331

Table 1. Percentage bacterial prevalence standard error (SE) at 10 different sampling


sites within the Oldman River Basin in 1999 and 2000, containing zero, low, medium, and
high animal manure units (AMU); in storm drains; and in urban settings.
Escherichia coli
Salmonella spp. O157:H7
Site Site classificationa n % SE % SE
1 Zero AMU density 21 19.0 8.8 4.8 4.8
2 Zero AMU density 24 13.0 6.9 8.3 5.8
3 Zero AMU density 17 18.0 9.5 0
4 Low AMU density 25 20.0 8.2 0
5 Medium AMU density 26 7.7 5.3 3.9 3.9
6 Medium AMU density 27 15.0 7.0 3.7 3.7
7 High AMU density 6 33.0 21.0 0
8 SD 4 25.0 25.0 0
9 SD 5 80.0 20.0 0
10 CL 22 4.6 4.6 9.1 6.3
Note: SD, storm drain samples; CL, sites within the City of Lethbridge that were not storm drain
samples.
a
AMU density zero, no confined livestock operations within a 5-km grid; low, mean AMU density
is between 17 and 166 within a 5-km grid; medium, AMU density between 166 and 315 within a 5-km
grid; and high, AMU density between 315 and 464 within a 5-km grid.

Results (Table 1) or (ii) downstream of high-density livestock opera-


tions. In contrast, water obtained from regions with high
Prevalence of Salmonella spp. and E. coli O157:H7 AMU were, generally, not contaminated with E. coli
In 1999, water was collected every 2 weeks throughout O157:H7, although prevalence of Salmonella spp. was high
the Oldman River basin from May through October. Salmo- in some cases (Table 1); animals in these regions were nor-
nella spp. were isolated from 14 of 468 water samples, for a mally confined and did not have direct access to surface wa-
prevalence of 3.0%. In 2000, water was collected every ter. No samples positive for E. coli O157:H7 were found
week from May through September. Salmonella spp. were within urban storm drain sites.
isolated from 74 of 961 samples, for a prevalence of 7.7%.
This represents a 2.6-fold increase in the prevalence of Sal- Manure production
monella spp. within the Oldman River basin from 1999 to Mean AMU values at sampling sites were 1655, 240, and
2000. During this period, the basin received less than aver- 394 for bovine, swine, and poultry operations, respectively.
age rainfall for the region and was suffering from drought Figures 1, 2, and 3, respectively, are maps detailing
conditions. Consequently, because of lack of water, not all E. coli O157:H7 prevalence, Salmonella spp. prevalence,
sites could be sampled consistently both in 1999 and in and interpolated surface of AMU for bovine operations, us-
2000; therefore, only bacterial prevalence at sites that were ing inverse distance weighting. Of 84 sampling sites, 10
consistently sampled in both 1999 and 2000 was compared. were not within 15 km of a confined feeding operation and
No significant annual differences were observed in the prev- had no interpolated AMU value; i.e., sites without bovine,
alence of Salmonella spp. (t = 1.15; one-tailed p = 0.1319) swine, or poultry AMU values were not included in the
at these sites (data not shown). On average, for 1999 and AMUpathogen prevalence analysis, while the remaining 74
2000, Salmonella spp. were detected in 88 of 1429 samples, sites were included. Locations of high bacterial prevalence
with a prevalence ( SE) of 6.2% 0.6%. A high proportion did not appear to be situated near large manure output
(26.3%) of the 38 samples from municipal storm drain sites sources (Figs. 1 and 2). Levels of aggregated manure output
contained Salmonella spp., and one of these had the greatest from local bovine (Fig. 3), swine, and poultry feeding opera-
Salmonella spp. prevalence (80.0% 20.0%; Table 1) ob- tions were not high at sampling sites that had a high bacte-
served in this study. rial prevalence (interpolated swine and poultry AMU maps
In 1999 and 2000, E. coli O157:H7 was isolated from 13 not shown).
of 1483 water samples for a prevalence (mean SE) of Consequently, no direct correlation between bacterial
0.9% 0.2%. As with the Salmonella spp. data, no signifi- prevalence and manure production from confined livestock
cant annual differences existed in the prevalence of E. coli feeding operations was identified. Bovine, poultry, and
O157:H7 (t = 1.451; one-tailed p = 0.0811) at sites that swine manure output associated with sampling sites was not
were consistently sampled both years (data not shown). On a significant predictor of Salmonella spp. or E. coli O157:H7
more than one occasion, E. coli O157:H7 was detected at an prevalence at sampling sites (Table 2) or among individual
irrigation outflow within the City of Lethbridge (Table 1). samples (Table 3).
However, most sites contaminated with E. coli O157:H7
were located near some form of agricultural activity (Fig. 1). Discussion
Further examination of these sites showed that they were lo-
cated (i) in areas with low to moderate livestock density This study is the first of its kind to isolate E. coli O157:H7

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332 Can. J. Microbiol. Vol. 49, 2003

Table 2. Associations between bacterial prevalence in surface water from sampling sites in
southern Alberta and manure output.
Type III likelihood
ratio test
Model Variable Estimate SE 2 p Error deviance/df
SALM Intercept 2.7698 0.5373 1.55
BOVAMU 0.0001 0.0001 0.2491 0.6177
SWNAMU 0.0005 0.0011 0.2267 0.6340
PLTAMU 0.0001 0.0008 0.0064 0.9365
ECOLI Intercept 4.8072 1.7548 0.6412
BOVAMU 0.0002 0.0003 0.2126 0.6448
SWNAMU 0.0036 0.0048 0.7043 0.4013
PLTAMU 0.0014 0.0022 0.4125 0.5207
Note: Poisson and binomial distribution models of Salmonella spp. and Escherichia coli O157:H7
prevalence, respectively, as functions of inverse distance weighting of regional manure output from
confined livestock feeding operations. SALM and ECOLI are the models of prevalence of Salmonella
spp. and E. coli O157:H7, respectively. BOVAMU, PLTAMU, SWNAMU are independent variables. is
the slope or parameter estimate for bovine, poultry, and swine animal manure units, respectively. df,
degrees of freedom; SE, standard error.

Table 3. Associations between bacterial presence and manure output (animal manure units) associated with surface-water samples of
southern Alberta.
Model likelihood Hosmer and Lemeshow
ratio test goodness-of-fit test
Bacteria Manure type B Wald p (df = 1) 2 p (df = 3) 2 p (df = 8)
Salmonella spp. Bovine 0.000 0.270 0.603 0.724 0.868 29.589 <0.001
Swine 0.001 0.231 0.630
Poultry 0.000 0.007 0.934
Escherichia coli O157:H7 Bovine 0.000 0.008 0.928 2.358 0.501 8.085 0.425
Swine 0.005 1.333 0.248
Poultry 0.000 0.002 0.922
Salmonella spp. All types 0.000 0.169 0.681 0.172 0.678 14.165 0.078
Escherichia coli O157:H7 All types 0.000 0.627 0.428 0.581 0.446 17.030 0.030
Note: df, degrees of freedom; B, logistic regression coefficient; Wald, statistic to test significance of B.

in surface waters collected from a Canadian watershed. Sys- than 15 years of age from Alberta that were diagnosed with
tematic sampling of surface water within the Oldman River hemolytic uremic syndrome (HUS) were likely infected with
basin in southern Alberta shows that it is often contaminated E. coli O157:H7 (Rowe et al. 1998). From 1987 to 1991, the
with E. coli O157:H7 and Salmonella spp. mean annual rate of VTEC infection for the province was
The presence of E. coli O157:H7 and Salmonella spp. in 12.1 cases per 100 000 compared with 2.0 per 100 000 in
water presents a risk to human health when the untreated wa- Scotland during the same period (Waters et al. 1994). Some
ter is consumed directly or indirectly. Our results identify of these cases might be the result of water contamination. In
E. coli O157:H7 and Salmonella spp. contamination in water our study, we repeatedly isolated E. coli O157:H7 from sur-
from surface drainage channels, some of which flows directly face waters within the Oldman River basin. Individuals who
into the Oldman and Little Bow Rivers and from storm drains consumed this untreated water or used it for recreational
and sewage treatment plants within the Oldman River basin. purposes would be expected to be at risk of infection (Pay-
While much of the public concern over these pathogens is re- ment et al. 1997; OConnor 2002).
lated to the consumption of contaminated food, concern has Khaleel et al. (1980) have suggested that a high density of
also been expressed over the contamination of water used for intensive animal rearing facilities could facilitate the con-
both drinking and recreational activities (Arvanitidou et al. tamination of surface water, resulting in large reservoirs for
1997; Polo et al. 1998; Sargeant et al. 2000). pathogenic bacteria. In 1992 in Swaziland, E. coli O157:H7
Michel et al. (1999) have shown that the risk of human in- caused one of the largest outbreaks of bloody diarrhoea in
fection by VTEC can be related to cattle density. In Canada, the world (Effler et al. 2001). At the end of a 3-month
Alberta has the greatest population of cattle and one of the drought, more than 40 000 people distributed over several
highest levels of gastroenteritis resulting from infection by hundred kilometres downstream of an agricultural district
Salmonella spp. and E. coli O157:H7 (Khakhria et al. 1996). were reported ill after runoff from heavy rains contaminated
Between 1991 and 1994, up to 77% of 205 children less food and water.

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Johnson et al. 333

Michel et al. (1999) have demonstrated an association be- tory in 1999 were isolated from poultry (Health Canada
tween human VTEC infections and cattle density in Ontario, 2000).
Canada. They suggested that regional manure spreading and In our study, a high number of the City of Lethbridge
water contamination might link animal density, a group-level storm drains were contaminated with Salmonella spp.
risk factor, and human infection, an individual-level outcome (Table 1). We note that much of the industry and a part of
(Valcour et al. 2002). Livestock manure is a reservoir for the population within the City of Lethbridge are engaged in
both Salmonella spp. and E. coli O157:H7 and is an impor- agricultural or related activities. It is not clear what contribu-
tant source of surface-water contamination. Cattle are recog- tion agriculture surrounding the city made to the observed
nized as reservoirs for E. coli O157:H7 (Dean-Nystrom et al. contamination. Further studies are needed to identify sources
1999), and cattle manure has been implicated as the source of contamination and to determine how city management
of contamination in the Walkerton outbreak that occurred in practices within agricultural regions might differ from those
Ontario, Canada (Kondro 2000). in large urban centres.
The Lethbridge region of southern Alberta is noted for The regional prevalence of E. coli O157:H7 was similar in
high cattle density (Fig. 3) as well as for one of the highest our study to that reported by McGowan et al. (1989) in their
levels of gastroenteritis in Canada, resulting from infection study of surface water; however, the numbers of Salmonella
by Salmonella spp. and E. coli O157:H7 (Khakhria et al. spp. we detected were similar to that seen in some studies
1996). This observation is similar to that of Michel et al. but were lower than that seen in others (Anonymous 1992;
(1999), whose work was done using geographical scaling at Efstratiou et al. 1998; Polo et al. 1998). We note that the
the census consolidated subdivision level. In our study, both prevalence of E. coli O157:H7 and Salmonella spp. through-
Salmonella spp. and E. coli O157:H7 were identified in sur- out the Oldman River basin was not uniform (Figs. 1, 2, and
face-water samples throughout the Lethbridge region, and it 3). While this variation in prevalence may, in part, be the re-
is likely that cattle contributed to this contamination. How- sult of various land management practices, other factors,
ever, when data from our study were examined using a such as time of sampling, geography, and weather, could
smaller geographical scale capable of identifying potential also influence these findings and other factors in any model
point sources, we found that surface water obtained close to developed.
large-scale animal feeding operations was not necessarily We attempted to model the effect of local AMU on water
more contaminated with zoonotic bacteria than were sam- contamination. However, the predictive strength of our
ples obtained far away from these operations; i.e., pathogen model was limited. Incorporation of other variables, such as
contamination of surface water, not including groundwater, bacterial transport and survival, into the model might im-
was not necessarily related to animal density when potential prove it. The velocity and direction of surface-water drain-
point sources of contamination were considered. age, the irrigation water flow, and the surface wind at time
We note that producers in southern Alberta have been of sample collection were not acknowledged within the
working to implement best management practices that mini- model but could strengthen it by providing a link between
mize the risk of water contamination within the region. Ani- the effect of local confined livestock operations and local
mals often are confined and do not have direct access to water contamination (these data were not available for this
surface water. Our data suggest that these practices may study). Environmental conditions at time of sampling, such
have a positive effect on water quality; i.e., no direct associ- as water temperature, pH, and nutrient concentration, could
ation of concentrated livestock facilities with pathogen prev- also be used to assess the potential for bacterial survival in
alence was detected in this study, suggesting that good water. These variables could influence bacterial concentra-
agricultural management practices can mitigate surface-water tion in water and may have influenced the ability of our
contamination within agricultural regions. These observa- model to identify an association between local manure load
tions, however, do not preclude that the spreading of manure and water contamination. In this study, we also assumed that
without sufficient treatment (e.g., composting) on fields may transport of manure beyond a 15-km radius of an animal
have contributed to the water contamination observed. Cows rearing facility would be prohibitively expensive and conse-
on pasture, wild deer, reptiles, and birds are reservoirs of quently, unlikely; this however, may not be the case.
pathogens, such as Salmonella spp. and E. coli O157:H7,
and may also have contributed to water contamination
(Bradley et al. 2001; Cizek et al. 1994; Fischer et al. 2001; Acknowledgements
Sargeant et al. 2000).
In our study, Salmonella spp. were more prevalent than This study was funded by Health Canada, the University
E. coli O157:H7 in water samples. Because Salmonella of Lethbridge, and the Alberta Agriculture Research Insti-
spp. are present at a low level in beef and dairy herds from tute. Water sample collection was by staff of Alberta Envi-
western Canada (Sorenson 2000; Van Donkersgoed et al. ronment (Water Management Division), Lethbridge Northern
1999), it is likely that other sources also contributed to the Irrigation District, Alberta Agriculture (Irrigation Branch),
water contamination that we observed. Van Donsel and Chinook Health Region, and the City of Lethbridge. R.
Geldreich (1971) have identified pigs as reservoirs for Sal- Hardin, Prairie Farm Rehabilitation Administration, pro-
monella spp. and note that they may contribute to sur- vided the animal manure unit data. A. Senthilselvan, Univer-
face-water contamination when runoff occurs. Domestic and sity of Alberta, assisted in the statistical modelling. We also
wild birds may also have been the source of some of the thank the Health Canada Salmonella Reference Laboratory
bacterial isolates obtained, e.g., 42% of Salmonella spp. sub- (Guelph, Ont.) and the CFIA Lethbridge Laboratory (Ani-
mitted to the Health Canada Salmonella Reference Labora- mal Diseases Research Institute) for their support of the pro-

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334 Can. J. Microbiol. Vol. 49, 2003

ject. Members of the research team are part of the Canadian Health Canada. 2000. 1999 Annual report of the Health Canada
Water Network. Salmonella reference laboratory. Health Canada Salmonella Ref-
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Johnson, R.P., Wilson, J.B., Michel, P., Rahn, K., Renwick, S.A.,
Gyles, C.L., and Spika, J.S. 1999. Human infection with
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