Methods Note/

Automated Time Series Measurement of

Microbial Concentrations in
Groundwater-Derived Water Supplies
by David W. Owens1 , Randall J. Hunt1,2 , Aaron D. Firnstahl3 , Maureen A. Muldoon4 , and Mark A. Borchardt5

Abstract
Fecal contamination by human and animal pathogens, including viruses, bacteria, and protozoa, is a potential
human health hazard, especially with regards to drinking water. Pathogen occurrence in groundwater varies
considerably in space and time, which can be difficult to characterize as sampling typically requires hundreds of
liters of water to be passed through a filter. Here we describe the design and deployment of an automated sampler
suited for hydrogeologically and chemically dynamic groundwater systems. Our design focused on a compact form
to facilitate transport and quick deployment to municipal and domestic water supplies. We deployed a sampler
to characterize water quality from a household well tapping a shallow fractured dolomite aquifer in northeast
Wisconsin. The sampler was deployed from January to April 2017, and monitored temperature, nitrate, chloride,
specific conductance, and fluorescent dissolved organic matter on a minute time step; water was directed to
sequential microbial filters during three recharge periods that ranged from 5 to 20 days. Results from the automated
sampler demonstrate the dynamic nature of the household water quality, especially with regard to microbial targets,
which were shown to vary 1 to 2 orders of magnitude during a single sampling event. We believe assessments of
pathogen occurrence and concentration, and related assessments of drinking well vulnerability, would be improved
by the time-integrated characterization provided by this sampler.

Introduction and related water availability. Fecal contamination

Human and animal pathogens are disease-causing
microorganisms that can affect suitability of the water for
certain use, and thus deteriorate groundwater resources
1 U.S. Geological Survey, Upper Midwest Water Science Center,
Middleton, WI 53562; dwowens@usgs.gov
2
Corresponding author: U.S. Geological Survey, Upper Midwest
Water Science Center, 8505 Research Way, Middleton, WI 53562;
(608) 821-3847; fax: (608) 821-3817; rjhunt@usgs.gov
3
U.S. Geological Survey, Laboratory for Infectious Disease and
the Environment, Marshfield, WI, 54449; afirnstahl@usgs.gov
4
Geology Department, University of Wisconsin-Oshkosh,
Oshkosh, WI, 54901; muldoon@uwosh.edu
5
USDA-ARS, Laboratory for Infectious Disease and the
Environment, Marshfield, WI, 54449; mark.borchardt@ars.usda.gov
Article impact statement: Using the automated sampler
described here, microbial concentrations in groundwater are shown
to vary significantly over short time periods.
Received March 2018, accepted August 2018.
by human and animal pathogens, including viruses,
bacteria, and protozoa, is a potential human health
hazard, especially with regards to drinking water sources
(e.g., Borchardt et al. 2012). Various environmental
factors (e.g., occurrence of groundwater recharge, pH,
temperature, salinity, UV light exposure) influence the
occurrence, fate, and transport of pathogens in the
subsurface. Moreover, there is a wide range of potential
pathogen sources, each with its own timing, magnitude,
and pathways of loading. Therefore, pathogen occurrence
in groundwater is expected to vary considerably in space
© 2018 The Authors. Groundwater published by Wiley Periodi-
cals, Inc. on behalf of National Ground Water Association.
This is an open access article under the terms of the Creative
Commons Attribution License, which permits use, distribution and
reproduction in any medium, provided the original work is properly
cited.
doi: 10.1111/gwat.12822

NGWA.org Groundwater 1
and time (e.g., Bradbury et al. 2013). However, even at
one location, pathogen characterization in groundwater
systems typically requires large volumes of water (e.g.,
600 to 1600 L) to be passed through a filter, which makes
characterization of temporal variability more difficult.
As a result, usually only one filter is collected. A more
comprehensive monitoring program that better represents
actual system dynamics would include consideration of
short-term and longer-term system variability.
The objectives of this study were to design, construct,
and test an automated sampler suited for hydrogeologi-
cally and chemically dynamic groundwater systems. Our
design focused on a compact form to facilitate trans-
port and quick deployment to municipal and domestic
water supplies. Although this discussion focuses on a sub-
set of possible microbial pathogens and indicators, we
believe these methods have applicability to a wide range
of microbe types present in groundwater systems.

Test Site
The test household was located in Door County,
Wisconsin (Figure 1), where the household well obtained
water from the upper fractured Silurian dolomite aquifer.
This aquifer is a regionally important, but vulnerable,
source of drinking water in northeast Wisconsin. In
southern Door County, dairy farming and associated crop
production comprise the primary land use and manure is
commonly applied to crop land. There are no communities
large enough to require municipal sewer systems and all
homes and commercial enterprises rely on septic systems
for on-site wastewater treatment. This region has a history
of bacterial contamination (Sherrill 1978) and occasional
“brown-water” events have also been observed in this
area, where household water supplies become temporarily
brown in color, odorous, and non-potable during large
recharge events. Because Door County soils are thin,
recharge to the dolomite aquifer can be exceedingly rapid
and can quickly transport surface contaminants into the
aquifer (e.g., Bradbury and Muldoon 1992; Muldoon and
Bradbury 2010).
The rapid response of the shallow aquifer is regional.
Previous work looking at recharge to the dolomite
aquifer (Muldoon and Bradbury 2010) tracked water-level
changes in four wells located in a four-county area, where
all wells had 10 to 20 feet of surficial sediment overlying
the dolomite aquifer. Water level changes were correlated
between wells, and all wells responded within 24 to 48 h
to a large precipitation or snowmelt event. This previous
work, and Sherrill (1978)’s observation that bacterial
contamination increases during recharge events, supported
the monitoring of a non-pumping “sentinel well” to
characterize timing and magnitude of recharge into the
larger aquifer system. Groundwater quality after recharge
events could then be sampled in domestic household water
supply.
This Technical Note describes results from an auto-
mated sampler deployed to one household; future publi-
cations will focus on the larger recharge characteristics
Figure 1. Location of automated sampler location, USGS
monitoring well KW183, and Casco, WI weather station
plotted on the state digital elevation model (in feet above
mean sea level).
and implications for well vulnerability. The household
well sampled during our study was 13.1 m (43 feet) deep
and the casing extended to 8.8 m (29 feet) as determined
by geophysical and borehole video logs (Peter Chase,
WGNHS, personal communication, 2016). The well con-
struction report for a replacement well (installed April 19,
2017) indicates that 6.7 m (22 feet) of surficial sediment
overlie the dolomite at this site. The sampler was deployed
January 18 through April 9, 2017.
Microbial Targets
Concurrent with auto-sampler deployment, a com-
panion study was being conducted in the same region
on the sources of fecal contamination in private wells.
Four microbial targets were frequently being detected in
the companion study: HF183 Bacteroides, ruminant Bac-
teroides, pepper mild mottle virus, and rotavirus group
A. In the auto-sampler work, we chose to target these fre-
quently detected microbial markers as well as commonly
reported total coliform concentrations.
Methods
To provide context for automated sampler design
decisions, a short description of a commonly employed

2 D.W. Owens et al. Groundwater NGWA.org

manual sampling method for pathogens and indicators
is provided, followed by a detailed description of
the automated sampler workflow. See Appendix S1,
Supporting Information regarding the methods used for
the microbial analyses.
Manual Microbial Sampling Protocol
Microbial samples from groundwater are typically
collected in two ways. Pumping wells are sampled by
connecting a filter to a wellhead tap while the pump was
running or piezometers are sampled by peristaltic or other
pump with the tubing sterilized between samples. Viruses
are concentrated in the field by dead-end ultrafiltration,
which has a pore size of approximately 30-kDa (Smith and
Hill 2009), or glass wool filtration (Millen et al. 2012).
Dead-end ultrafiltration is typically easier to implement
in the field and has been used to detect a range of
pathogens. Moreover, glass wool filtration requires pH
conditioning when groundwater has pH values of 7.5
or higher to ensure virus pathogens adhere to the filter;
dead-end filtration does not have this same requirement.
It is recommended that filters be shipped on ice to a
laboratory within 48 h of sample collection, where they
are backflushed, followed by concentration of the filtrate.
The final concentrated sample volume from each filter
(typically around 4 mL) is stored at −80 ◦ C until nucleic
acid extraction.
Automated Microbial Sampler
The automated sampling was performed using a
custom-designed, automated, refrigerated large-volume
virus filtration system (Figure 2). The sampler was
installed in the basement of a single-family dwelling
that used a domestic well. The household water supply
was tapped at the water pressure tank and fed to an
(a)
datalogger and
telemetry
enclosure
Legend
A – water pressure tank (WPT)
B – solenoid ball valve
C – rotary flowmeter
D – manual valve
E – ultra filter
F – quick disconnect tube connector
G – flow-through chamber for water quality sensor
H – water quality sensor
I – flexible discharge tube to drain/sump
J – pressure transducer
Figure 2. (a) Schematic diagram of automated sampler. (b) Photograph looking into automated sampler refrigerated
enclosure.
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array of dead-end hemodialysis ultrafiltration filters that
were accessed sequentially using solenoid ball valves and
datalogger controls. Remote telemetry to the 12-V battery
backed up datalogger allowed operators to monitor current
conditions, adjust operational parameters and provide
automated data retrieval. Sampling was initiated and
monitored using a smartphone. In the work shown here,
this design allows sample coverage of entire recharge
events (as identified using a United States Geological
Survey [USGS] Climate Response Network well; see
Figure 1) without locating field personnel at the household
site over the course of the whole event.
The sampling system monitored flow rates with
several inline rotary flow meters. A flow rate was set
using a valve on the system intake; the initial flow rate
was set to around 1.2 ± 0.1 L/min. Eight solenoid ball
valves were used to sequentially direct flow between
the eight dead-end ultrafiltration filter units. The flow
was directed through the filters sequentially based on the
volume of water passed through a filter. Target volumes
could be changed for the current or future filters during
the sampling event. This allows the user to optimize
the event sample coverage as the event progresses. A
pressure transducer measured backpressure in the filter
system, an indication of filter clogging and/or system
failure. When a threshold pressure of 345 kPa (50 psi)
was met the system would be shut down by closing the
main valve into the sampler. If glass-wool filters were
used, the automated sampler design described here could
be modified to accommodate pH conditioning (e.g., used
by Corsi et al. 2014 for sampling surface water). This
sampler used dead-end ultrafiltration thus pH-conditioning
was not needed. Handling of the filters after collection
from the automated sampler followed the same protocol
as the manual approach described above.
(b)
D.W. Owens et al. Groundwater 3
 Water moving through the sampler was also moni-
tored using water-quality sensors, including temperature,
specific conductance, pH, nitrate, chloride, fluorescent dis-
solved organic matter (FDOM), and turbidity using a
multi-probe water-quality sonde and a flow-through cham-
ber. Measurements were made every minute and stored
on the sampler’s datalogger. Water-quality data collected
were available real-time using the two-way telemetry of
the sampler. The purpose of this monitoring was to char-
acterize the dynamics and conditions of the household
system during time periods when pathogen samples were
taken. Because only periodic calibration was performed
during the study, reported values are considered to approx-
imate actual analyte concentrations. During initial testing,
degassing of the water in the flow-through chamber caused
bubble artifacts in results from the water-quality sensors
(discussed below); orientating the sonde more vertically
reduced, but did not eliminate, bubble formation. Water
was continually flushed through the flow-through cham-
ber at a rate of approximately 0.6 L/min. This resulted in
a constant relatively low-flow rate discharge that required
constant disposal using gravity drainage into a floor drain.
Because the sampler is designed for installation in domes-
tic households it included an emergency shutdown capa-
bility triggered by the occurrence of water in the bottom of
the sampler. The sampler was constructed using a small
refrigerator; therefore, filters could be refrigerated until
they were removed to increase holding times.
Approach for Sampler Testing
Hydrologic conditions used to determine when
to activate microbial samplers were tracked using
online resources. The regional groundwater system was
monitored using real-time data available from a USGS
monitoring well (site number/name: 443535087345401
KW-25/24E/34-0183; USGS, 2017).
The local conditions in the household water supply
were monitored on a minute time step using low-flow
rates routed through the water-quality sonde flow-through
chamber. When a target recharge event was identified in
the monitoring well hydrograph, the smartphone was used
to open the valve to the first filter. The sampler program
then monitored the flow rate until the user-specified
volume passed through the filter, at which point the valve
to the first filter was shut and the valve to the second
filter was opened. This continued until all filters were
used or the user canceled the sampling event. Filters
were collected every 1 to 3 days during the testing to
minimize holding times and maintain continual coverage
over the sampling period. During collection, the sampler
was paused, filters with quick disconnect couplers were
replaced, used filters placed in a plastic bag, sealed,
and shipped to the analytical laboratory. Protective latex
gloves were worn throughout filter collection to prevent
contamination.
Results and Discussion
Our initial discussion focuses on insights gained from
the entire January to April 2017 sampling period; a
4 D.W. Owens et al. Groundwater
0.0
Depth to water below land surface (meters)
0.2
hand measurement
datalogger measurement
0.4
0.6
0.8
1.0
1.2
Event #1 Event #2 Event #3
1.4
1.6
7-Jan 21-Jan 4-Feb 18-Feb 4-Mar 18-Mar 1-Apr 15-Apr
Figure 3. Hydrograph from U.S. Geological Survey Moni-
toring Well KW183 showing changes in water level during
the study period.
March to April 2017 snowmelt event within this period
is examined in more detail. All water quality data shown
here are provided by Owens et al. (2018).
Entire Sampling Period
Hydrologic Conditions and Sampling Approach
Data from the USGS monitoring well shows the
period had multiple recharge events (Figure 3). The
sampler was run to characterize household water quality
during three of these recharge events including: Event
#1, had 7 sequential filters collected during a relatively
large recharge event January 18 to 23; Event #2, had 8
sequential filters collected during a small recharge event
February 28 to March 4; and Event #3, had 26 sequential
filters collected during a large recharge event March 21
to April 9. The last of these three events is discussed in
detail in the next section. After sampling was initiated, a
filter received water until a user-specified threshold was
reached; the amount of water passed through a filter was
750 L (Events #2 and #3) and ranged between 600 and
990 L (Event #1) as the filter collection schedules were
being refined. No filter clogging was observed during the
study.
Geochemical Time Series
Although the water quality sonde data were only
used to provide background characterization and identify
potentially anomalous conditions in the household water
system, results are included here for illustrative purposes.
Water quality measured by the sampler’s sonde show
fluctuation operating on the order of days and weeks
(Figure 4). This is perhaps not surprising because the
sonde is sampling a mixed average water quality of
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 1000000

Total coliform MPN/10 liters

100000
Human Bacteroides gc/L
Pepper mild mottle virus gc/L
10000

Microbial concentration
Rotavirus group A (NSP3 gene) gc/L

1000

100

10

Figure 4. Household water supply quality over the entire 0.1

study period as measured by the automated sampler
sonde (deg ◦ C = degrees Celsius; mg/L = milligrams per
liter; RFU = relative fluorescence units; microS/cm = micro-
Siemens per centimeter).
the aquifer (i.e., both slow transport/bulk flow and fast
transport/low yield contributions that transport pathogens;
Hunt et al. 2010). In addition, the effects of recharge in
the groundwater system are seen in the geochemical time
series when the recharge events are relatively large (e.g.,
Event #1 and #3; Figure 4). However, individual analytes
can move in similar directions, be lagged, or move
inversely, likely reflecting the complexity inherent to
multiple sources and dual domain transport in the shallow
fractured rock aquifer. Water temperature, however, was
relatively constant throughout the period the sampler was
deployed.
Event #1 Event #2 Event #3
0.01
7-Jan 21-Jan 4-Feb 18-Feb 4-Mar 18-Mar 1-Apr 15-Apr
Figure 5. Results of microbial analyses performed on dead-
end ultrafilters collected from the automated sampler during
three events. Note all data are plotted on a log scale. All five
microbial targets were analyzed for each filter; ruminant
Bacteroides was not detected during the sampling. Non-
detections are not shown; when a target organism was absent
a gap between time series markers occurs (MPN = most
probable number; gc/L = gene copies per liter).
Table 1
Measured Daily Precipitation, Casco, WI (http://
www.enviroweather.msu.edu/weather.php?
stn=lux)

Precipitation Precipitation
Microbial Water Quality Time Series
Date (2017) (inches) (mm)
The household microbial water quality was extremely
variable, and characterized by appreciable changes in March 21 0 0
microbial presence/absence and concentration over short March 22 0 0
periods of time (Figure 5, note the log scale). In all cases March 23 0 0
but total coliform, microbes analyzed were characterized March 24 0.64 16.3
by presence/absence between filters collected sequentially March 25 0.14 3.6
March 26 0.35 8.9
in time; when present a microbial concentration is shown March 27 0.06 1.5
but when absent a gap between time series markers occurs. March 28 0 0
Ruminant Bacteroides was not found in any sample during March 29 0.02 0.5
the period. Total coliform, found in water from the well March 30 0.40 10.2
in all but one filter, can vary 1 to 2 orders of magnitude March 31 0 0
during sampling events (Figure 5). It should be noted April 1 0 0
that the symbols in Figure 5 (and later on Figure 7) April 2 0.11 2.8
April 3 0.11 2.8
reflect the time that a filter was collected for ease of
April 4 0.39 9.9
plotting; the value measured for a filter can only reflect April 5 0 0
the average microbial concentration over the time water April 6 0 0
passed through the filter. April 7 0 0
April 8 0 0
March 21 to April 9 Snowmelt Event

Hydrologic Conditions
A spring 2017 snowmelt period was chosen for
detailed description, where the event extended from
March 21 to April 9. During this period rain was
noted at the Casco, Wisconsin weather station (15 km
from the sampler location; Figure 1) during three distinct
periods, March 24 to 27, March 29 to 30, and April 2
to 4 (Table 1). All three periods were associated with
increasing groundwater levels (Figure 3) in the USGS
Climate Response Network monitoring well (14 km away
from sampler location; Figure 1).

NGWA.org D.W. Owens et al. Groundwater 5

 30 Filter# 820 250

Total coliform
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 24 25 26 Human Bacteroides gc/L
temperature/nitrate/chloride/FDOM Pepper mild mottle virus gc/L
25 800 200
Total coliform MPN/10 liters

specific conductance
20 780
150

100
15 760
sonde
calibration

50
10 740

PMMV concentration
Human Bacteroides
100 0
5 720

10
0 700
21-Mar 23-Mar 25-Mar 27-Mar 29-Mar 31-Mar 2-Apr 4-Apr 6-Apr 8-Apr 10-Apr
temperature (deg C) nitrate-N (mg/L) chloride (mg/L) FDOM (RFU) specific conductance (microS/cm) 1

Figure 6. Household water supply quality as measured 0.1

by the automated sampler sonde during Event #3
sonde (deg C = degrees Celsius; mg/L = milligrams per 0.01
liter; RFU = relative fluorescence units; microS/cm = micro- 19-Mar 26-Mar 2-Apr 9-Apr 15-Apr
Siemens per centimeter).
Figure 7. Results of microbial analyses performed on dead-
end ultrafilters collected from the automated sampler during
Event #3. Note that total coliform is plotted on an arithmetic
scale and human Bacteroides and pepper mild mottle virus
Geochemical Time Series
(PMMV) are plotted on a log scale. All five microbial targets
Sonde data are summarized as again having broad were analyzed for each filter; ruminant Bacteroides and
trends rather than sharp changes (Figure 6). Two Rotavirus group A were not detected during the Event
trends are apparent during Event #3 that demonstrate #3 sampling. Non-detections are not shown; when a target
organism was absent a gap between time series markers
an inverse relation between specific conductance and occurs (MPN = most probable number; gc/L = gene copies
nitrate and chloride; specific conductance increased per liter).
then decreased over the sampling period while nitrate
and chloride decreased then increased during the same
period (Figure 6). The lack of synchronicity may reflect
Best Practices and Potential Limitations
enhanced flushing of carbonate-rich water in the aquifer
This sampler design was successful in its ability to
dolomite matrix and smaller contributions from fractures
provide continual sampling coverage of the household
that contain surface-derived nitrate and chloride. Temper-
water supply. This work was particularly notable for its
ature was again consistent at or just below 10 ◦ C over
focus on the more difficult objective of characterizing
the period.
the water quality of a domestic household water supplies
Some water chemistry analyte time series appear to
over a multi-week period; previous research evaluating
have artifacts from degassing and associated bubble for-
domestic household wells typically employed a single
mation/release during this period (e.g., FDOM; Figure 6).
sampling event to assess pathogens in domestic wells
Moreover, recalibration of the sensors on the sonde on
(e.g., Borchardt et al. 2003). From the large variability in
April 6 also resulted in a change in the sonde reported
time series data, one suggested best practice that results
values (e.g., specific conductance, nitrate; Figure 6), more
from this work is to limit extrapolation of manual samples
variable time series after recalibration (FDOM; Figure 6),
of groundwater to a single snapshot of current conditions
and an extended recovery period that persisted over a day
and not as representing a longer-term, time-integrated
(chloride Figure 6). As noted previously, these results are
microbial system.
perhaps best considered in a more general sense of house-
Best practices for deploying the sampler include four
hold water-quality conditions rather than precise concen-
primary topics. First, the field personnel should have
trations of the analytes.
uniform procedures that protect against contamination
during switching of the filters, such as using color coded
Microbial Time Series filter ends, double bagging, and new sample gloves for
In contrast to the broad trends of the sonde data, time each collection trip. Second, it is critical that the flow
series of total coliform and virus results from 26 sequen- rates reported by the sampler are accurate because they
tial filters again show a dynamic system where microbial are used to automate the filter switching and are also
concentration and occurrence is variable over time, and used for calculating microbial concentrations. Third, the
microbial targets other than total coliform were char- field personnel should assess the vertical orientation of the
acterized by presence/absence between filters collected water quality sonde to reduce bubble adherence to sensors.
sequentially in time. Moreover, total coliform can vary And fourth, although refrigeration extends holding times,
1 to 2 orders over the event sampled (Figure 7—note the the sampling schedule should be designed to reduce, to the
log scale on the left axis and arithmetic scale on the right extent practicable, the time between a filter being filled to
axis). when it is processed in the lab.

6 D.W. Owens et al. Groundwater NGWA.org

 Current limitations of the sampler design presented
here include:
• Long-term deployment of the water-quality sonde
appeared to result in probe drift that was corrected
during recalibration. Therefore, the water quality char-
acterization should be considered qualitative unless
more rigorous quality assurance is employed (e.g., more
frequent calibration). However, inclusion of a water
quality sonde in the sampler provides a cost-effective
geochemical context for the water supply over time.
Such insight is important for choosing microbial sam-
pling periods, which helps ensure representative results
while lowering the cost of microbial sampling.
• Most household water systems are under pres-
sure, therefore artifacts of bubble formation during
depressurization/degassing are expected to occur unless
steps are taken to keep the flow-through chamber
pressurized.
• Inadvertent failure to restart the flow to the flow-through
chamber after sonde calibration is not flagged by the
datalogger so gaps in the sonde time series can occur
(such as seen in March, Figure 4).
• High accuracy in measurement of flows through the
system can also be difficult to attain as the flows are near
the low end of the operating range of many flow meters.
Orienting the flowmeter so the exit point elevation is
higher than the entrance point elevation might increase
accuracy, and will be tested as part of ongoing sampler
improvement.
• Similar to any long-term water monitoring, the amount
of water discharged can be large. At some sites
sump pumps or floor drains may not be available.
However, the sampler shown here is suitable for
deployment outdoors during non-freezing conditions
using an outside spigot.
• When scheduling filter collection to reduce holding
times, the number of visits to a site increases. This
will increase the cost of field personnel and homeowner
disruption.
The sampler also has the more general limitations of
microbial characterization in groundwater systems includ-
ing: lack of information on the source, poor definition of
contributing area of the well, and difficulty anticipating
transport and lags in transport in the shallow fractured
bedrock of the study area. Therefore, longer sampling
periods are needed (and more filters/analysis required) to
ensure a target system is adequately characterized.
Conclusions
The results presented here demonstrate that microbe
occurrence and concentration time series can be efficiently
characterized in the field. These results represent those of
shallow fractured aquifers with rapid travel times. In this
setting, the sampler documents highly variable domestic
household water quality over time. Characterization of
groundwater quality using single snapshot sampling can
NGWA.org
be expected to leave longer-term characterization of
pathogen occurrence and magnitude highly uncertain.
Therefore, deploying samplers as described here can be
critical for understanding pathogen source and transport,
as well as microbial dynamics, in aquifer settings.
Acknowledgments
This work was funded by both the State of Wisconsin
Department of Natural Resources through the State
Groundwater Research Advisory Council and the U.S.
Geological Survey Cooperative Matching Funds Program.
We thank the homeowners who graciously allowed us
to use their household systems to develop and test the
sampler design presented. Davina Bonness, Travis Engels,
from Kewaunee County and Forrest Kalk (USGS) are
thanked for project assistance and filter retrieval. We
also thank Pete Lenaker (USGS) for assistance with an
early design of the sampler, and Jason Smith (USGS) for
sonde calibration and monitoring well data collection. Pete
Chase (Wisconsin Geological and Natural History Survey)
is thanked for geophysical logging of the household
well. Reviews from Anita Anderson and James Walsh
(Minnesota Department of Health) and two anonymous
reviewers greatly improved the manuscript. Any use of
trade, firm, or product names is for descriptive purposes
only and does not imply endorsement by the U.S.
Government.
Authors’ Note
The authors do not have any conflicts of interest or
financial disclosures to report.
Supporting Information
Additional supporting information may be found online
in the Supporting Information section at the end of
the article. Supporting Information is generally not peer
reviewed.
Appendix S1. Methods used for microbial sample pro-
cessing and analysis.
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