Environmental Pollution 120 (2002) 771–778

www.elsevier.com/locate/envpol

Escherichia coli and total coliforms in water and sediments at

lake marinas
Youn-Joo Ana,*, Donald H. Kampbellb, G. Peter Breidenbachc
a
Oak Ridge Research Associateship Program, US Environmental Protection Agency, Office of Research and Development, National Risk Management
Research Laboratory, PO Box 1198, Ada, OK 74820, USA
b
US Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory, PO Box 1198,
Ada, OK 74820, USA
c
ManTech Environmental Research Services Corp., Robert S. Kerr Environmental Research Center, 919 Kerr Research Drive, Ada, OK 74820, USA

Received 2 November 2001; accepted 28 February 2002

‘‘Capsule’’: Escherichia coli levels in surface water increase due to resuspension of sediment-bound bacteria by recrea-
tional boating activities at lake marinas.

Abstract
Escherichia coli, a fecal coliform, and total coliforms were monitored between September 1999 and October 2001 in five marinas
at Lake Texoma, located on the Oklahoma and Texas border. The general trend was that densities of E. coli were lower in the
summer season due to the lower loading of fecal material into Lake Texoma and the ecological conditions of the lake, such as more
vigorous grazing by protozoa and less viability of E. coli at an elevated temperature. The densities of total coliforms greatly
increased in the summer. E. coli levels increased with depth, and the bottom water samples had higher densities of E. coli mainly
due to their association with particles. There was a direct relationship between amount of gasoline sold, which was related to
recreational boating activity, and the resuspension of E. coli. This indicated that recreational boating activity in lake marinas may
have resuspended bottom sediments with bound E. coli, and the presence of E. coli in marinas was not an indication of recent fecal
contamination. E. coli were detected in the largest densities at the boat dock points, followed by the gasoline filling station, and
marina entrance. In addition, enumeration of bacteria in bottom sediment showed that the densities of E. coli and total coliforms in
sediment were much higher compared to those in lake water. # 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Escherichia coli; Fecal coliform; Total coliform; Marina; Resuspension

1. Introduction and also indicates the potential for the presence of

Escherichia coli (E. coli) is the most common member
of fecal coliform bacteria, indigenous to the intestinal
tract of humans or other warm-blooded animals
(Dufor, 1977). The US Environmental Protection
Agency (EPA) recommended that E. coli is a better
indicator of fecal pollution than fecal coliform for pur-
poses of evaluating ambient fresh water quality
(USEPA, 1986). The presence of E. coli in lake water
indicates that the water was contaminated by fecal
material of humans or other warm-blooded animals,
* Corresponding author. Present address: Department of Environ-
mental Science and Engineering, Ewha Womans University, Seoul,
South Korea. Tel.: +82-2-3277-4238; fax: +82-2-3277-3275.
E-mail address: anyjoo@ewha.ac.kr (Y.-J. An).
0269-7491/02/$ - see front matter # 2002 Elsevier Science Ltd. All rights reserved.
PII: S0269-7491(02)00173-2
pathogenic organisms. The source of E. coli con-
tamination in surface water includes municipal waste-
water discharges, septic leachate, agricultural or storm
runoff, wildlife populations, or nonpoint sources of
human and animal waste.
The decay of fecal coliform bacteria is influenced by
several environmental factors. The rate of die-off
increases with increasing temperature (Flint, 1987), ele-
vated pH, and higher dissolved oxygen levels (Curtis et
al., 1992). Other factors include solar radiation (Game-
son and Saxon, 1967; Gannon et al., 1983; McCam-
bridge and McMeekin, 1984), predacious
microorganisms such as protozoa (McCambridge and
McMeekin, 1984; Sinclair et al., 1993), lack of nutrients
(Van der Steen et al., 2000) and salinity (Davies and
Evison, 1991).
772 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778
Lake Texoma is located on the border of southern
Oklahoma and northern Texas. It has 93,000 surface
acres, and is a focus of the recreation, and farming
industries in the region. There are potential stressors
around the Lake Texoma watershed that may cause
fecal pollution in the lake. Examples are septic tanks,
chicken farms, wildlife refuges, etc. Fecal material can
flow into the lakes through runoff. Fecal coliforms in
lake water are predominantly removed by adsorption
onto particles and subsequent sedimentation; thus,
viable fecal coliform bacteria usually accumulate with
sediment (Gannon et al., 1983). Higher densities of fecal
coliform in sediment than in water are well documented
elsewhere (Crabill et al., 1999; Goyal et al., 1977; Irvine
and Pettibone, 1993). At marinas, there is a potential
for resuspension of bottom sediments with bound fecal
coliform bacteria through boating activities, which
churn the water, resulting in increased fecal coliform
suspension in lake water.
In this paper, we report the seasonal occurrence of E.
coli and total coliforms in lake marinas from a 2-year
monitoring study of Lake Texoma. We also evaluated
the influence of recreational boating activity on the E.
coli densities in lake marinas, and their spatial distribu-
tion between lake water and sediments. Bacteria levels
in lake water were monitored from September 1999
through October 200. Their enumeration in sediments
was measured in the fall of 2001.
2. Materials and methods
2.1. Field sampling
Lake water samples were obtained at five different
marinas from September 1999 to October 2001. Sam-
pling locations are shown in Fig. 1. Samples were col-
lected at 1-foot depths at locations identified as marina
entrance, gasoline filling station, and boat dock. In
addition, samples were collected at 8 feet from the sur-
face and at 1 foot from the bottom at the gasoline filling
stations. Water samples were collected through a 14 inch
diameter polyethylene tube connected to a peristaltic
pump and placed in sterile 100-ml vessels with sodium
thiosulfate (0.1 ml of a 1.8% solution per 100 ml capacity).
Paired water and sediment samples were collected
from eight stations at five marinas in September and
October of 2001. Sediment samples were taken follow-
ing water sample collections at the locations. Sediment
samples were collected using a small modified Peterson
dredge sampler (LaMotte, Code 1097) that was rinsed
with lake water. Samples were collected from surface
sediment (approximated 0–1 cm depth) and placed into
sterile plastic bottles. All samples collected were placed
in a cooler with blue ice and delivered the same day to
the analytical laboratory.
Ten percent or more of the samples were collected in
duplicate. Quality assurance measures performed on the
set of samples included duplicates, known analytical
quality controls (AQCs), and blanks.
2.2. Laboratory experiments
Total coliform and E. coli in water samples were
determined using the Colilert systems that can simulta-
neously detect and enumerate both organisms (Eaton et
al., 1995; Edberg et al., 1990). The density units are
expressed with most probable number (MPN) per 100
ml. Reverse osmosis (RO) purified water was used as a
negative control and RO water with one drop of a log
phase culture of E. coli ATTC strain #15597 was used as
a positive control. The limit of quantification for this
method is 1 MPN/100 ml.
Total coliform and E. coli in sediment samples were
enumerated by spreading suitable dilutions of the sedi-
ments on eosin methylene blue (EMB) plates that select
for Gram negative bacteria. An aliquot of sediment (the
equivalent of 1.5 g of dry wt.) was placed in a 40-ml
glass vial and 15-ml of sterile RO water was then added
to the vial. The vials were closed and rolled on a low
profile roller (Stovall, Life Science, Inc.) at 8 rpm for 1 h
to suspend the sediment; this is a 1:10 dilution. Further,
10-fold dilutions were made from 100:1 through 1000:1.
Half (0.5) ml aliquots of these dilutions were placed on
the surface of EMB plates in duplicate and the aliquot
spread over the surface of the agar with a sterile L-
shaped glass rod. The inoculated EMB plates were
incubated for 2 days at 36  C and the number of colo-
nies was counted on plates with between 30 and 300
colonies present, utilizing a colony counter. The number
of colonies with a green-metallic sheen were counted
and recorded as the colony forming units (CFU) of E.
coli per gram soil of dry weight and the total number of
colonies present recorded as the CFU per g of dry
weight. When the number of CFU on duplicate plates
varied by greater than 33%, the dilution was replated.
3. Results and discussion
3.1. Seasonal occurrence of E. coli and total coliforms in
lake water
Seasonal differences of E. coli densities were found in
Lake Texoma. Fig. 2 shows the densities of detectable
E. coli during September 1999 to October 2001. E. coli
was present in marina water during all seasons in Lake
Texoma indicating that the water was contaminated by
fecal material of humans or other warm-blooded ani-
mals. Occurrence of E. coli in most of the samples was
greater than about 70% as shown in Fig. 2 (see circles).
The mean densities varied from a minimum of < 1
 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778 773

Fig. 1. Lake Texoma showing the sampling locations. Stations 1–3, 4–6, 7–9, 10–12, and 13–15 are located in A, B, C, D, and E marinas,
respectively.

MPN/100 ml to a maximum of 179 MPN/100 ml in all
samples. Generally, monthly geometric mean (n510) of
E. coli was lower in the summer season at marinas. This
effect may be explained by two factors; lower loading of
fecal material, and the ecological conditions of the lake
(e.g. viability of E. coli, grazing by protozoa). First,
runoff during rainfall is a major factor in the loading of
fecal materials into the lake. The positive relationship of
fecal coliform densities in river or lake water with runoff
or discharge was previously reported (Barbe and Fran-
cis, 1995; McDonald et al., 1982). Fig. 3 compares the
monthly total rainfall at the Lake Texoma basin with E.
coli densities during the monitoring period, showing the
general positive relationship of E. coli densities in lake
water with rainfall data. As concluded from drought
conditions during summer 2000, as shown in Fig. 3, E.
coli densities were lower in this period. A wet season
after the drought resulted in the higher loading of E. coli
774 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778
Fig. 2. Escherichia coli densities at marina waters in Lake Texoma during September 1999 to October 2001. Bars and circles (o) represent the mean
densities, and percent occurrence of bacteria each sampling dates, respectively.
and subsequent high E. coli levels in lake water. In
addition, higher loading of nutrients with runoff could
help the growth of E. coli, if any growth occurs.
Second, microbial survival and rate of decay seems to
depend on the ecological conditions of the lake during
the season. Temperature may be the most important
single environmental factor with other factors including
protozoan predation, solar radiation, algae growth, etc.
Die-off of fecal coliform bacteria was apparently more
rapid at an elevated temperature (Flint, 1987), or per-
haps grazing by protozoa was enhanced so it appeared
they were more viable in cold water. The abundance of
protozoa would reduce the bacterial community (Sin-
clair et al., 1993). Decay of bacteria by solar radiation
has been well documented (Gameson and Saxon, 1967;
Gannon et al., 1983; McCambridge and McMeekin,
1984). Algal photosynthesis can affect bacteria counts
by increasing pH and DO (Pearson et al., 1987; Van der
Steen et al., 2000). Higher algal concentrations during
the summer were reported in Lake Texoma marinas (An
et al., 2000).
Fig. 4 shows the seasonal occurrence of total coli-
forms monitored from September 1999 to October 2001.
Total coliforms were present in all samples. The den-
sities of total coliforms greatly increased in the summer
since non-fecal coliforms can grow at an elevated tem-
perature (Baudisova, 1997). This trend agrees with the
previous observation in a river where total coliform
counts increased in the summer (Baudisova, 1997).
3.2. E. coli occurrence in depth and sampling site
Fig. 5 shows the ratios of E. coli density in surface
water (1 foot from the lake surface) to bottom water (1
foot from lake bottom). About 57% of all ratios in
Fig. 5 are less than 1, indicating that bottom water
samples have higher densities of E. coli, mainly due to
their association with particles. Fecal coliform bacteria
in lake water are usually found to be associated with
colloid sized small particles. About 90% of fecal coli-
forms were associated with particles that ranged from
0.45 to 10 mm (Auer and Niehaus, 1993).
Another factor may be the lower water temperature
of bottom water since fecal coliforms are reported to be
more viable in cold water (Flint, 1987). Pearson et al.
(1987) reported fecal coliform levels were lower at high
temperature, high dissolved oxygen and high pH. How-
ever, densities of fecal coliform in marina waters were
not always lower in the surface water where temperature
and DO are higher. This indicated that one of the fecal
coliform sources could be human recreational activity.
Boating activity in lake marinas can resuspend bottom
sediments with bound fecal coliform bacteria, which
may also be a causative factor. Pettibone et al. (1996)
 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778
Fig. 3. Comparison of monthly total rainfall with Escherichia coli levels in Lake Texoma during September 1999 to October 2001.
observed that ship traffic in Buffalo River, New York,
increased the levels of fecal coliform bacteria in the
water column by resuspension of sediments. Boating
activities were previously reported in Lake Texoma
during the summer months (An et al., 2000, 2001,
2002). Fig. 5 also shows the average gallons of gasoline
sold per month in 2001 at four marinas (A, C, D, and
E), which are related to the recreational boating activity
in Lake Texoma. There was a positive correlation
between the gasoline sold (proportional to boating
activity) and the bacteria resuspension, which showed
the frequent resuspension of E. coli by recreational
activity in summer months. This phenomenon also
suggests the presence of E. coli in marinas is not always
an indication of recent fecal contamination, although
there is a possibility of direct dumping of untreated
waste from boats into the lake during recreation
activities.
E. coli levels were compared at the surface for three
sampling locations. They were detected in larger den-
sities at boat dock points (38%), followed by the gaso-
line filling station (23%) and marina entrance (17%).
Higher density of E. coli in boat dock samples could be
caused by wildlife, such as ducks that lived around
boat-launching facilities, as well as resuspension of
sediment-bound bacteria by active boat traffic at these
areas. Extensive waterfowl populations were reported to
775
have a direct impact on concentrations of fecal coli-
forms in surface waters (Henderson, 1968).
3.3. E. coli levels in bottom sediments
Table 1 shows the total coliforms and E. coli densities
measured in sediments and overlying lake waters. The
E. coli densities ranged from 3.50104 to 5.00105
CFU per gram of sediment as dry weight, which showed
that the densities in sediment are greater than the den-
sities in Lake Texoma water like many other water
bodies. The mean ratio of total coliform densities to E.
coli densities in sediment was calculated to be 5.1  3.1
at Lake Texoma marinas in the fall months and there
was a positive correlation between both densities in
sediment (r=0.874).
A similar attempt to correlate E. coli densities in
water and in sediment was not successful. This indicated
that the E. coli densities in water are not a good indi-
cator for densities in sediments.
4. Conclusions
The 2-year monitoring study for E. coli and total
coliforms at marinas in Lake Texoma showed seasonal
occurrence in lake water. Densities of E. coli were
776 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778

Fig. 4. Total coliform bacteria at marina waters in Lake Texoma from September 1999 to October 2001.

Table 1
Escherichia coli and total coliform densities in sediments and lake water at Lake Texoma, September and October 2001

Marina Sampling station Date Sediment (CFU/g dry sediment) Lake water (MPN/100 ml)

Total coliform E. coli Total coliform E. coli

A 01 10/1/01 2.81106 5.00105 >2419.2 <1

A 03 10/1/01 1.69106 3.80105 >2419.2 <1
B 06 9/12/01 1.77105 3.50104 >2419.2 2
C 08 9/12/01 8.60105 3.60105 >2419.2 <1
C 09 9/12/01 2.73105 9.30104 >2419.2 <1
D 11 9/12/01 6.40105 1.50105 >2419.2 6.25
D 12 9/12/01 7.40105 6.00104 >2419.2 1
E 15 10/1/01 1.60106 4.60105 >2419.2 <1

generally lower in the summer season possibly due to (1)
the lower loading of fecal material during summer, (2)
more vigorous grazing by protozoa, and (3) less viability
in warm water. The densities of total coliforms greatly
increased in the summer at elevated temperatures.
Higher ratios of E. coli in surface water to bottom water
were detected at marinas during the summer months,
indicating the resuspension of viable sediment-bound
bacteria by recreational boating activities in lake mar-
inas. Resuspension of E. coli during the boating season
was directly related to the amount gasoline sold, which
would be directly related to the recreational boating
activity. Higher densities of E. coli were detected at boat
dock points, followed by the gasoline filling stations and
were lowest at the marina entrance. E. coli and total
coliform counts in sediment were much higher com-
pared to those in lake water.
Acknowledgements
The US Environmental Protection Agency through its
Office of Research and Development funded and man-
aged the research described here through in-house
efforts. It has not been subjected to Agency review and
therefore does not necessarily reflect the views of the
 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778
Fig. 5. Escherichia coli ratios in surface water to bottom water in Lake Texoma marinas. Circles (*) show the ratio and a line shows the average
values of gasoline sold at four marinas (A, C, D, and E marina) per month in 2000. Gasoline data at B marina was not available.
Agency, and no official endorsement should be inferred.
This research was supported in part by an appointment
to the Postgraduate Research Participation Program at
the National Risk Management Research Laboratory
administered by the Oak Ridge Institute for Science and
Education through an interagency agreement between
the US Department of Energy and the US Environ-
mental Protection Agency. We thank Mike Cook in US
EPA, Ada, OK for his assistance with sampling. Jason
Masoner in US Geological Survey assisted with map
preparation.
References
An, Y.-J., Kampbell, D.H., Sewell, G.W. (2000) Water quality of three
marinas in Lake Texoma. Fall meeting of American Geophysical
Union, San Francisco, CA, Supplement to Eos, Transactions,
American Geophysical Union, 81, 48, F385.
An, Y.-J., Kampbell, D.H., Sewell, G.W. (2001) Occurrence of methyl
tert-butyl ether (MTBE) at five marinas in Lake Texoma. The 222nd
National Conference of American Chemical Society, Chicago, IL,
Division of Environmental Chemistry Preprints of Extended
Abstracts, 41, 2, 478–484.
An, Y.-J., Kampbell, D.H., Sewell, G.W., 2002. Water quality at five
marinas in Lake Texoma as related to methyl tert-butyl ether
(MTBE). Environ. Pollut. 118, 331–336.
777
Auer, M.T., Niehaus, S.L., 1993. Modeling fecal coliform bacteria. I.
Field and laboratory determination of loss kinetics. Wat. Res. 27,
693–701.
Barbe, D.E., Francis, J.C., 1995. An anlysis of seasonal fecal
coliform levels in the Tchefuncte River. Wat. Res. Bull. 31,
141–146.
Baudisova, D., 1997. Evaluation of Escherichia coli as the main indi-
cator of faecal pollution. Wat. Sci. Tech. 35, 333–336.
Crabill, C., Donald, R., Snelling, J., Foust, R., Southam, G., 1999.
The impact of sediment fecal coliform reservoirs on seasonal water
quality in Oak Creek, Arizona. Wat. Res. 33, 2163–2171.
Curtis, T.P., Mara, D.D., Silva, S.A., 1992. Influence of pH, oxygen,
and humic substances on ability of sunlight to damage fecal coli-
form in waste stabilization pond water. Appl. Environ. Microbiol.
58, 1335–1343.
Davies, C.M., Evison, L.M., 1991. Sunlight and the survival of enteric
bacteria in natural waters. J. Appl. Bacteriololy 70, 265–274.
Dufor, A.P., 1977. Escherichia coli: the fecal coliform. In: Hoadley,
A.W., Dutka, B.J. (Eds.), Bacterial Indicators/Health Hazards
Associated With Water. ASTM, Philadelphia, pp. 48–58.
Eaton, A.D., Clesceri, L.S., Greenberg, A.E., 1995. Standard Method
for the Examination of Water and Wastewater, nineteenth ed.
APHA, AWWA, WEF, Washington, DC.
Edberg, S.C., Allen, M.J., Smith, D.B., Kriz, N.J., 1990. Enumeration
of total coliforms and Escherichia coli from source water by the
defined substrate technology. Appl. Environ. Microbiol. 56, 366–
369.
Flint, K.P., 1987. The long-term survival of Escherichia coli in river
water. J. Appl. Bact. 63, 261–270.
Gameson, A.L., Saxon, J.R., 1967. Field studies on effect of daylight
on mortality of coliform bacteria. Wat. Res. 1, 279–295.
778 Y.-J. An et al. / Environmental Pollution 120 (2002) 771–778
Gannon, J.J., Busse, M.K., Schillinger, J.E., 1983. Fecal coliform dis-
appearance in a river impoundment. Wat. Res. 17, 1595–1601.
Goyal, S.M., Gerba, C.P., Melnick, J.L., 1977. Occurrence and
distribution of bacterial indicators and pathogens in canal com-
munities along the Texas coast. Appl. Environ. Microbiol. 34,
139–149.
Henderson, J.M., 1968. Enteric disease criteria for recreational waters.
J. Sanit. Engr. Div., Proc. ASCE SA6, 1253–1276.
Irvine, K.N., Pettibone, G.W., 1993. Dynamics of indicator bacteria
populations in sediment and water near a combined sewer outfall.
Enviorn. Technol. 14, 531–542.
McCambridge, J., McMeekin, T.A., 1984. Effect of solar radiation
and predacious microorganisms on survial of fecal and other bac-
teria. Appl. Environ. Microbiol. 41, 1083–1087.
McDonald, A., Kay, D., Jenkins, A., 1982. Generation of fecal and
total coliform surges by stream flow manipulation in the absence of
normal hydrometeorological stimuli. Appl. Environ. Microbiol. 44,
292–300.
Pearson, H.W., Mara, D.D., Mills, S.W., Smallman, D.J., 1987. Phy-
sico-chemical parameters influencing faecal bacterial survival in
waste stabilization ponds. Wat. Sci. Tech. 19, 145–152.
Pettibone, G.W., Irvine, K.N., Monahan, K.M., 1996. Impact of ship
passage on a bacteria levels and suspended sediment characteristics
in the Buffalo River, New York. Wat. Res. 30, 2517–2521.
Sinclair, J.L., Kampbell, D.H., Cook, M.L., Wilson, J.T., 1993. Pro-
tozoa in subsurface sediments from sites contaminated with aviation
gasoline or jet fuel. Appl. Environ. Microbiol. 59, 467–472.
USEPA, 1986. Ambient water quality criteria for bacteria—1986.
EPA440–584–002. US EPA, Washington, DC.
Van der Steen, P., Brenner, A., Shabtal, Y., Oron, G., 2000. Improved
fecal coliform decay in integrated duckweed and algal ponds. Wat.
Sci. Technol. 42, 363–370.