WATER RESOURCES RESEARCH, VOL. 33, NO.

4, PAGES 639-648, APRIL 1997

Bacteriophage and microsphere transport in saturated porous

media: Forced-gradient experiment at Borden, Ontario
Roger C. Bales, Shimin Li, 1 and T.-C. Jim Yeh
Department of Hydrologyand Water Resources,Universityof Arizona, Tucson

MelissaEl Lenczewski
2 and CharlesP. Gerba
Department of Soil, Water and EnvironmentalScience,Universityof Arizona, Tucson

Abstract. A two-wellforced-gradientexperimentinvolvingvirus and microsphere

transportwas carriedout in a sandyaquiferin Borden,Ontario, Canada.Virus traveledat
leasta few metersin the experiment,but virusconcentrations at observationpoints 1 and
2.54 rn awayfrom the injectionwell were a small fraction of thoseinjected.A simplified
planarradial advection-dispersionequationwith constantdispersivity, coupledwith
equilibriumand reversiblefirst-ordermasstransfer,wasfoundto be adequateto simulate
the attachmentand transportprocess.During the experimenta short-durationinjectionof
high-pH water was also made, which causeddetachmentof previouslyattachedviruses.
For simulatingthis detachmentand associatedtransport,the sametransportand mass-
transferequationswere used;but all rate parameterswere varied as groundwaterp H
changedfrom 7.4 to 8.4 and then back to 7.4. The physicochemical parametersobtained
from fitting breakthroughcurvesat one samplingwell were usedto predict thoseat
anotherwell downstream.However, laboratory-determinedparametersoverpredicted
colloidremoval.The predictedpattern and timing of biocolloidbreakthroughwas in
agreementwith observations, thoughthe data showeda more-dispersebreakthroughthan
expectedfrom modeling.Thoughclearlynot an equilibriumprocess,retardationinvolving
a dynamicsteadystate betweenattachmentand detachmentwas neverthelessa major
determinantof transportversusretentionof virus in this field experiment.

Introduction vection and dispersion[Baleset at., 1991; Gerba et at., 1991;

Harvey, 1991]. Advection dependson groundwatervelocity.
The occurrenceand transportof human entericvirusesand Dispersiondependson velocityand aquifer heterogeneityand
bacteria in and through groundwater have been a long- is scaledependent.Attachment and detachmentrates are sen-
standingpublicconcern.Questionsof virustransportare cen- sitive to groundwaterchemicalconditions,such asp H, ionic
tral to groundwaterwellhead protection. Distancesbetween strength,and the compositionof the porous media [Gerba,
virussourcessuchas septictanksand drinkingwater wells are 1984;Bales et al., 1993], and in many casesare the most im-
typicallyset on the basisof empiricalrelations.Quantitative portant factorscontrollingbacteriaand virus transport.Inac-
estimatesbased on site-specificcharacteristicsor transport tivation of virusesdependsstronglyon temperature [Yateset
modelinghave not been usedfor settinggroundwaterprotec- al., 1987] and is typicallyslowcomparedto the rates of advec-
tion standards.
tion, attachment,and detachment[Baleset al., 1995].Accurate
Physicaland mathematicalmodelsfor describingthe fate of
predictionof virus transportthroughporousmedia near their
biocolloids(i.e., virusandbacteria)in porousmediahavebeen
sourcethereforeoften dependssolelyon the correctevaluation
suggestedfor morethan a decade[e.g.,Vilker,1978;Vilkerand of the rates at which viruses attach to or detach from the
Burge,1980;Funderburget al., 1981;Grosset,1985;Corapcioglu
porousmedium surfaces.
and Haridas, !984, 1985; Yateset al., 1987; Matthesset al., 1988;
This researchwasaimedat evaluatingmathematicaldescrip-
Harveyand Garabedian,1991]. However, applicationof these
tions of the chemical and microphysicalfactors controlling
modelshassufferedin part from a lack of systematicfield and
biocolloid transport through porous media. We undertook a
laboratoryresearch.Accumulationof experimentaldata and
validationof existingor newlydevelopedmodelswith data are forced-gradient
fieldexperimentandsubsequent mathematical
essential. modelingwith threemainquestionsin mind:(1) Canwe model
The major factorscontrollingvirusand bacteriafate in sub- virus (bacteriophage)
and microspheretransportat the field
surfaceporousmedia are attachmentto and detachmentfrom scaleusingthe samefew microscale parametersas havebeen
the porousmediumsurfaces,growthand inactivation,and ad- usedin the laboratory?,(2) How do colloidattachmentto and
detachmentfrom aquifer media affecttransportrelative to that
of a conservativesolutein a heterogeneous system?,and (3)
1Nowat ArizonaDepartmentof Environmental Quality,Phoenix. How does a disturbancein groundwaterp H facilitate further
2Nowat AmwayCorporation, Ada, Michigan.
transport of previously retained bacteriophage or micro-
Copyright1997 by the American GeophysicalUnion. spheres.We usedbacteriophageand microspheresrather than
Papernumber97WR00025. animal virus or bacteria becauseour primary aim was model
0043-1397/97/97WR-00025 $09.00 testing.With thesemodelcolloidswe havebetter experimental
639
640 BALES ET AL.: BACTERIOPHAGE TRANSPORT

test involvinginjection well 2 and a pesticidetransport test.

EXT PP2 ML5 ML4 IN J2 Mackay et at. [1994] reported results of an earlier organic
solutetransportexperimentat the site.
J?Before
• _•_

0 The extractionwell waspartiallypenetratingwith a screened

lengthof 5 m, slot sizeof 0.5 mm, and a total lengthof 6.2 m.
The supplywell wasalsopartiallypenetrating,with a screened
lengthof 1.5 m, slot sizeof 0.5 mm, and total lengthof 3.7 m.
The injection well was partially penetratingwith a screened
6
length of 1.35 m and 0.25-mm slot size.The inner diameter of
7
the extraction,supply,and injectionwells was 51 mm.
..
There were 20 water table observation wells scattered

8 6 4 2 0
around the test site, and 18 piezometersaligned with three
straight lines in the radial direction of the extraction well
Distance, rn within 30 m of the extraction well. These observation wells and
Figure 1. Portion of well systemfor field test at Borden, piezometerswere designedand installedby previousresearch
Ontario: cross-section view. Screened sections of extraction
groups.Each observationwell wasmade of 25-38 mm diame-
(EXT), partiallypenetrating(PP2), and injection(INJ2) wells ter PVC pipes, fitted with a length of nylon screenedopen
for this experiment are shaded.To the left of EXT was a sectionmade accordingto the estimateddrawdownat each
secondinjection(INJ1) well and a partiallypenetratingwell
point. Piezometerswere similar, but had a consistent0.2-m
that were used in the concurrentpesticide-transportexperi-
ment (not shown).ML4 andML5 were multilevelmonitoring length of nylon-screened open section.Piezometerswere de-
wells;only the depthssampledare shown.Water table levels signed to measure drawdown at a "point," while water table
during and before experimentare alsoshown. observationwells were designedto measurean averagedraw-
down over a finite vertical distance.
The water table was monitoredwith a tape each day during
control and detection.We alsousedmultiple colloidsin order the experimentat both piezometersandobservationwellsafter
to comparetheir relativebehavior. the extractionwas started. More-frequent observationswere
made at a few piezometerscloseto the extractionwell at the
beginningof the extraction,sincedrawdownat early timeswas
Site Description very fast. There were rain eventsduringthe experiment;mea-
The test site waswithin an inactivesandquarry at the Ca- surementswere made immediatelyafter each rain to monitor
nadianForcesBase,Borden,Ontario.The aquiferat the siteis the effect on the water table.
unconfinedand composedof clean,well-sorted,fine- to medi- At the extractionwell a centrifugalpump was installedto
um-grained sand with distinct horizontal bedding evident maintaina largehydraulicgradientat the site.A flowmeterwas
[Mackayet at., 1986;Sudicky,1986].The thickness of the aqui- used to measure the instantaneous flow rate. The extracted
fer is about 9 m, and the groundsurfaceis almosthorizontal. waterwasdischargedto a pond about200 m awayfrom the site
On the bottomof the aquiferis a thick, siltyclaydeposit.The to avoid any disturbanceto the local flow field. Water with-
drybulksoildensityis 1.81gcm-3, with a solidsdensityof drawn from the supplywell was reinjected to the aquifer
2.71gcm-3 andspecific surface
areaof 0.8m2 g- • [Mackay et throughthe two injectionwells.The flow rate at the extraction
at., 1986]. Organic-carbon content (0.03%) and cation- wellwas30L min-•. Themaximum flowrateatthesupply well
exchange
capacity
(0.5meqg-•) arelow,witha clay-size
frac- was 5 L min -•. Flow rates were 2 L min -• at INJ2 and 3 L
tion near zero (reported as 0-15%). Dominant ions in the min- • at INJ1.
aquiferwereCa2+ andHCO•-,withp H of 7.4-7.5.
The measuredhydraulicconductivities of 1279samplesfrom
Methods
32 cores measuredwith a falling head permeametervaried
over more than 1 order of magnitudeand indicateda horizon- The study,carriedout in July1991,consisted of two sequen-
tal layered structure;the overall geometricmean of the hy- tial experiments;in the first, attachmentof colloidsto aquifer
draulicconductivity
(corrected
to 10øC)is 7.2 x 10-s m s-• media dominated, and in the second, detachment as well as
[Sudicky,1986]. attachmentwas important.For the first (attachment)experi-
The natural hydraulicgradientat the site hasbeen observed ment the injectionmixturecontainedphagesPRD-1 and M-l,
to rangefrom 0.0035to 0.0054throughthe year [MacFartaneet polystyrenemicrospheres,and NaC1 as a conservativetracer.
at., 1983];the bestestimateof an averagehydraulicgradientis The phageand microspheresolutionswere madewith 5 mL of
0.0043 [Mackayet at., 1986;Sudicky,1986].Averagednatural 0.1-/•mmicrospheres
(9.82x 10•3microspheres mL-•), 10mL
groundwater velocity
was0.09m d- • [Mackay etat.,1986].The of PRD-1(1.24x 10•2plaque-forming units(pfu)mL-•), 10
effectiveporositywas estimatedto be approximately0.3, that mL of M-1 (10mpfumL-•) and3 L of distilled
water.For the
is, on the order of 90% of the estimatedtotal porosity[Mackay second(detachment)experimentthe injectionsolutionwas
et at., 1986].Both slightandsignificantseasonalchangesin the high-pH water with no phageor microspheres. Sodiumphos-
naturalflow directionhad beenobservedby differentresearch- phate(Na2HPO4)wasusedto preparethe high-pHbuffer,and
ers [MacFartaneet at., 1983].The water table was found to be NaOH wasusedto adjustthepH first to 8.6 and later to 10.0
verysensitiveto rains,fluctuatingbetween0.5 and 1.0 m from in the injectate,after we found a p H of 8.6 wasnot adequate
groundsurfacein responseto the rains duringour field test. to raisethe p H at the injectionwell becauseof dilution.
The whole well systemconsistsof one extractionwell, one PRD-1 wasobtainedfrom J. Hsieh (Departmentof Micro-
supplywell, and two injectionwells (Figure 1). Two field ex- biologyand Immunology,College of Medicine, Universityof
perimentswere conductedat the sametime: a virus-transport Arizona,Tucson).Bacteriophage PRD-1 is an icosahedral(20

10 ' I . i . i . • . ,
a. attachment
Microsphere
o 4
---\M-1 I read using an OlympusBH-2 microscope(OlympusOptical
311
C•tivity
0 , 2I • 4I • 6I , B
I , 10 0 5
Time, hr
Figure 2. Injectionhistoryat injectionwell 2 (a) for attach- pfu mL-• of PRD-1 andM-1 andplacedin a clean,sterile
ment and transportexperimentand (b) for detachmentand polypropylenecentrifugetube.The tubewasplacedin an 11øC
transportexperiment.Note backgroundconductivitywas 311 water bath, and sampleswere taken and assayeddaily. Levels
taScm-•. Covaluestakenas1.1 x 108mL-• for microsphere, of
1.7 x 106 pfu mL-• for PRD-1,and1.6 x 105pfu mL-• for
M-1.
triangularfaces)lipid phagewith an averagediameterof 62 nm
[Olsenet al., 1974]. Its isoelectricpoint in calciumphosphate Results
buffer(10-4 M Ca) isbetween
pH 3 and4 [Bales
etal., 1991].
PRD-1 is a member of the Tectivirdae,a group of double-
strainedDNA somaticvirusescomposedof an outer protein
coat that enclosesa protein-lipidmembranevesicle[Bamford
et al., 1981]. The lipid is not exposedon the surfaceof the
phage.Two proteinsmakeup the proteincoatof the virus43.1
and 34.3 kda in molecularmass.Both appearas homomultim-
erswhere the subunitsare stronglyboundtogether.The amino
acid sequenceof protein P5 appearsin a collagenlikemotif.
Additional information on the structureof PRD-1 is given by
Bamfordet al. [1995].
M-1isah•at-resistant
bacteriophage
thatwasisolated
inour
laboratory from Tucson sewage.The M-1 virus is a typical
T-evendouble-stranded phagewith a head,tail, baseplate, and
tail fibersbelongingto the genusMycoviridae.It is a somatic
phagethat attachesto the cell wall of the host bacteriaby a
baseplate. The head of the phageis an icosahedron,elongated
by one or two extrabandsof hexamersof 85 x 110nm. Its shell
consistsof 5-nm-diameter capsomerscomposedmainly of
three proteinsof molecularweightsof 46 and 960 aminoacids.
The contractiletail (25 x 110 nm) is composedof a tube,
sheath,and connectingtail with a collar and whiskers,with a
complexbase plate with tail fibers.A detailed descriptionof
the compositionof the structuralproteinsis givenby Frankel-
Conratet al. [1988]. Attachment to inanimatesurfacesis usu-
allybytheheador tail.Thep Hiep of thephageis4-5 [Gerba,
1984].
Microsphereswere 100-nmmonodispersed unchargedpoly-
styrenebeadscontainingyellow-greenfluorescentdye (couma-
rin excitationmaximum,458 nm; emissionmaximum,540 nm),
from Polysciences, Inc., Warrington, Pennsylvania.
The host bacteria were Salmonellatyphimurium LT2 for
PRD-1 and Escherichiacoli (ATTC 15597) for M-1. The
phageswere assayedby the pfu methodsas describedby Ad-
ams [1959].All virusassays were performedin duplicate.Rep-
licate assaysof a singlesamplefor phagedifferedby an average
of 25% for all samples.A detailed descriptionof the phage
assayis given by Baleset al. [1991]. The microspheres were
enumeratedon polycarbonatemembranefilters (Nuclepore,
BALES ET AL.: BACTERIOPHAGE TRANSPORT 641
! , i i .
Pleasanton,California) with a pore size of 0.05 tam and a
b. detachment
diameter of 25 mm using epifluorescentmicroscopy.Filters
E were renderedblack by soakingfor 24 hoursin a solutionof 2
of Irgalan blackdissolvedin 1 L of 2% aceticacid,then rinsed
with distilledwater and dried [Hobbieet al., 1977]. Each sam-
ple wassonicatedfor 20 min to break up aggregates; then 1-10
mL of samplewas vacuumfiltered to depositcolloids.Filters
were then placed on glassslideswith a drop of glyceroland
Co., Tokyo, Japan) fitted with epifluorescence. We observed
the emissionfrom a 0.1-tammicrosphererather than the mi-
0 crosphereitself. Ten representativefieldswere counted.
10 15 20
To determine the inactivationrate of PRD-1 and M-l, ap-
Time, hr
proximately
50mL of Bordengroundwater
wasmixedwith104
both PRD-1 and M-1 decreased about 25% over the 2-week
period.We thereforeassumedthat over the courseof the field
experimentthere was no significantlossof bacteriophagedue
to inactivation,consistentwith past experience[Yahyaet al.,
1993].
Experimental Breakthrough Curves
Figure 2 showsthe injectionhistoryof PRD-1, M-l, micro-
sphere,and electricalconductivityobservedat the injection
well duringthe attachmentexperiment.The assayedphageand
microsphereinjection concentrationsfluctuated during the
first injection period. For example,the PRD-1 concentration
was1.7 x 106 pfu mL-• at the beginning anddecreased to
3.6 x 105 neartheendof theinjection.
Electricalconductivity
and chlorideconcentrationgraduallydecreasedto their back-
groundvalues.Becausethe fluctuationsof the differenttracers
in the injectatewere not the same,causesof the fluctuations
could not be attributed to a singlefactor. Insufficientmixingin
the carboy and in the injection pipeline, fluctuationsin the
injectionwater pumpingrate, and error in the enumerationof
phagesand microsphereswere all possiblecontributors.
Breakthroughcurves(Figure 3) obtainedfrom the first ex-
perimentshowedstrongretentionof colloidsbut alsoevidence
of remobilization.Virus andmicrosphereconcentrations at the
two samplingwells were so low that complete PRD-1 and
microspherebreakthroughcurveswere observedonly at levels
6 and 8 of well MIA. That is, concentrationsat other sampling
pointswere near or below the detectionlimit during break-
through. In this context a "complete" breakthrough curve
means a concentrationor electric conductivityversus time
curve startingfrom the lower limit of concentrationmeasure-
ment, increasinggraduallyto a high concentrationvalue, then
decreasinggraduallyback to the lower limit of the measure-
ment. The M-1 breakthroughcurvewasonly completeat level
6 of MIA and level 8 of ML5. Intermittent appearancesof
virusesandmicrospheres were evidentat almostall of the eight
pointssampledin the two wells.Peak concentrationvaluesand
times to peak, used to estimate the groundwatervelocity in
each layer, are given in Table 1. The highestPRD-1 concen-
trations
were440pfumL-• at MIA and12pfumL-• at ML5.
Microspheresappeared at relatively higher concentrations
than the two typesof viruses.The highestmicrospherecon-
centrations were 2.8 x 105 mL -• at MIA and 4.7 x 104 mL -•
at ML5.
642 BALES ET AL.: BACTERIOPHAGE TRANSPORT

M-1 Microsphere PRD-1 pH

6 8.5

8.0

7.5

7.0

4 i ß ß i ß i ß i , , , , 4 8.5 I I i

I
xl-3 8.0

•2 -
7.5
o)1

7.0

5 ß I ß i ' I ' i i i I i i 8.5 i i i

14
8.0

L•3 -I 000
¸ ¸%00 •0 ••o
7.5
031

0 ,,•m.m•,, • •,',..• ,,..--, ,.--•.,,,,.•.••..•,,..v,,•,,.,, ,,.,, ,..x,, '-- ---'' '"'"• • ' 7.0

5 i i i i i i 4 8.5

14
.•
•3 o o
o

i 8.0

7.5

0 •' C_ ----::-
' - --•':
'- ',-=---:; ' •:•:' -_::n::•O•c:• __-- 2 7.0 ' • ' ' ' '
0 5 10 15 0 5 10 15 10 15 20 0 5 10 15

Days Days Days Days

Figure 3. Injectatebreakthroughs andp H at samplinglevels6 and 8 of wells ML4 and ML5 for both
experiments. The first injectionwasat time = 0 and the secondwasat time = 11 days.Circlesare colloid
concentrationsand lines are conductivity.Virus concentrationsare in plaque-formingunits per milliliter,
microsphere is numberper milliliter,andconductivityis microsiemons
per centimeter.Samplesbelowdetec-
tion limit are plotted as C = 10ø.

The times of appearanceof the PRD-1 peakswere 3 to 6 arrival of the breakthroughsindicate retardation of phage
times thosefor the conductivitypeaks.For microspheresthe transport involvingreversibleretention and remobilization.
lag wasmuchless,abouthalf a day (Table 1). The M-1 major The PRD-1 andM-1 peakconcentration valuesat the sampling
peak alsolaggedthe conductivitypeak time. These delaysin well 0.94 m downstreamfrom the injection well were only

Table 1. Peak Information After the First Injection

Conductivity? PRD- 1$ M-lõ Microsphere

Sampling Depth, Time, Value, Time, Concentration, Time, Concentration, Time, Concentration,
Point m p H* days /xScm-• days pfu mL- • days pfumL- 1 days 103 mL- 1
ML4-3 2.3 7.5 0.8 1140 5.0 2 0.7 8 1.0 33
ML4-4 2.5 7.6 0.5 1050 5.0 8 1.2 5 1.2 9
ML4-5 2.7 7.7 0.5 1000 4.8 6 1.3 2 0.8 13
ML4-6 2.9 7.7 0.5 1070 1.8 440 1.7 26 1.0 275
ML4-8 3.3 7.6 0.5 1090 3.0 126 4.3 3 0.8 10
ML5-4 2.5 7.5 3.2 680 5.2 2 3.5 5 2.0 47
ML5-6 2.9 7.6 2.8 680 0.3 3 ...... 2.7 17
ML5-8 3.3 7.7 4.7 840 5.2 12 2.0 1 5.0 2

*AveragebeforepH of the groundwaterwasmodified.

?Thehighest
electrical
conductivity
valueat theinjectionwellwas1160/xScm-•;background
was311/xScm-•.
$Thehighest
PRD-1concentrationat theinjectionwellwas1.7 x 106 pfumL-•
õThehighest
M-1 concentration
at theinjectionwellwas1.6 x l0s pfumL-•.
Thehighest
microsphere concentration
at theinjection
wellwas1.1 x 108mL-•.
 BALES ET AL.: BACTERIOPHAGE TRANSPORT 643

Table 2. Peak Information After the SecondInjection

Conductivity? PRD-1 M-1 Microsphere

Sampling Depth, Time, Value, Time, Concentration, Time, Concentration, Time, Concentration,
Point m p H* days /•S cm-• days 103pfumL- • days 103pfumL- • days 103mL- •
ML4-3 2.3 8.0 12.5 650 12.0 0.3 11.5 1.3 12.0 0.5
ML4-4 2.5 8.5 11.7 1020 11.8 100 11.8 12.9 11.8 340
ML4-5 2.7 8.2 11.8 890 11.8 135 11.8 4.2 11.0 7.6
ML4-6 2.9 8.4 11.7 1060 12.2 1260 11.8 23.9 12.5 4.0
ML4-8 3.3 8.2 12.2 890 12.7 314 12.3 4.0 12.7 101
ML5-4 2.5 8.1 15.0 470 12.5 0.002 14.2 0.002 11.0 3.2
ML5-6 2.9 8.2 14.2 600 15.7 3.7 14.0 0.3 14.2 2.6
ML5-8 3.3 8.0 11.8 560 15.8 0.1 15.8 0.013 15.7 0.5

*The highestp H recordedat the injectionwell was 8.3.

]-Thehighest
conductivity
at theinjection
wellwas1654/•Scm-•.

0.01% of the injectionconcentrations;at the secondsampling smallportion of the Borden aquiferoff the main experimental
well, 2.54 m downstreamfrom the injectionwell, theywere less field site was extensivelysampled for stochasticanalysisof
than 0.001% of the injectate.The conservativetracer concen- spatial variability of hydraulic conductivity[Sudicky,1986].
trationsat the two samplingwellsshowedno similardecrease, This task aimed at deriving generic information about the
ruling out dispersionas the main causeof the attenuationof heterogeneityat the Borden site. Using this genericinforma-
colloid concentrations. tion, many studieshavetestedmacrodispersion concepts(i.e.,
The electricalconductivity breakthroughcurves(Figure 3) assumingthe aquifer is homogeneous)in the main portion of
showedan initial breakthroughof the injectate,followedby a the aquifer. Resultsof the studiesdemonstratedthat the ge-
secondunexplainedpeak; this secondpeak was apparently neric informationwill be usefulif and only if the tracer plume
from another chloridesourceremainingfrom prior salt injec- has traveled for a distance of hundreds to thousands of meters
tion at the site. [e.g., Sudicky,1986]. For small scaletracer testssuchas our
Breakthroughcurvesfrom the secondexperiment(Figure 3) field experiment,one mustrely on a site-specific descriptionof
showedsignificantremobilization,followed by further trans- the heterogeneity[e.g.,Yeh, 1992;Yehet al., 1995b].However,
port and retention.There wasno evidentretardationof viruses a detailed three-dimensional site characterization of our small-
and microspheres observedrelativeto the electricalconductiv- scalefield sitewasnot available.As a result,our analysisof the
ity. The times that peaksarrived at most pointsin ML4 and forcedgradienttestrelieson a simplifiedone-dimensional flow
ML5 werenearlythe samefor the three colloids(Table 2). The and transportapproach.
highestPRD-1 concentrationscausedby increasingthe p H of When molecular diffusion is much smaller than mechanical
groundwater were1.3 x 106 pfumL-• at ML4 and3.7 x 103 dispersionand rV = constant,where r and V are the distance
pfumL-• at ML5, 1000timesthatobserved duringtheattach- from the injectionwell and velocityat r, Bear [1979] obtained
mentandtransporttest.For M-1 theywere2.4 x 104 pfu a one-dimensionaladvection-dispersion equationfor a plane
mL-• at ML4 and2.7 x 102pfu mL-• at ML5. The highest radial transportproblem.Sincethe injectionrate in our field
microsphere
concentrations
were3.4 x 105mL-• at ML4 and test was much smaller than that of the extraction rate, we can
3.2 x 103mL-• at ML5. Fluctuations
of p H and electrical assumethat the conditionrV = constantwasprobablymet for
conductivitywere also observedduring the injection (Figure most of the r values and that the one-dimensionalequation
2), but the overalltrend wasconsistent with the experimental applies. When viral inactivation is not significant, one-
adjustment,and thereforeconsideredto be due to fluctuations dimensionaltransportcanbe describedby a combinedkinetic-
in water supplyrate. Note the two-stepgradual rise in p H equilibriumtwo-sitemodel [Cameronand Klute, 1977;Baleset
corresponding to the increaseinp H firstto 8.6 and then to 10.0 al., 1991]:
in the injectate.Figure3 showsthe observedpH changesat the
OC OSi 0S2 0 OC 0
two multilevelsamplingwells.Values at mostwellsincreased (])
up to 0.5pH unit. Note thatp Hs in water withdrawnfrom the
OuC
injectionwell were lower than thosein the injectateowingto
p H bufferingby the soil and dilution. s, = iCp,C (2)
Samplingat the extractionwell showednonzerophagecon- 0S2
centrationsin only one sample.Dilution at the extractionwell pb•-= OklC- pbk2S2 (3)
undoubtedlycausedconcentrationsto drop below the detec-
tion limit.
D = adu (4)
Modeling Phage and Microsphere Transport where C is the concentrationof virusesin groundwater,S 1 is
Mas-Plaet al. [1992],Yehet al. [1995a],andMcCarthyet al. concentrationof thosevirusesattachedon the fast (equilibri-
[1996]showed
thatcorrect
estimation
of physicochemical
pa- um) sitesof soil surfaces,S2 is concentrationof thoseviruses
rametersfor microorganism transportin the field demandsan attachedon the slow(non-equilibrium)sitesof soil surfaces,u
accuratedescriptionof groundwatervelocity.This impliesthat is the interstitial groundwatervelocity,D is the dispersion
three-dimensionalmodeling and detailed characterizationof coefficient,
Kpl is the equilibriumdistribution
coefficient
for
the experimentalfield site are necessary.During the 1980sa the equilibriumsitesin cubiccentimetersper gram, kl is the
644 BALES ET AL.: BACTERIOPHAGE TRANSPORT

ML4-6 ML4-8
k2= (k2i t<tx
k22 tx--<or
t --<t>
tx tx+to
+ to (7)
-4 ½

O•ø
O -5 5 tx =
•0
xdx
u(x)
(8)

-• o o The second1 in the abovesubscriptsstandsfor the rate and

equilibriumconstantsvalid before the chemicaldisturbance
-7 7 , I, , , •
arrivedat (time tx) or after it passes(time tx + to) by a given
point;the 2 representsthe variablevalid duringthe periodof
chemical disturbance.
Curvefittingwasusedto estimateand evaluatethe chemical
-3 ß Microsphere- Microsphere
parametersof the phageandmicrosphere transportat the field
o
_

site.Numericalsolutionsto the governingequationswere first

o
derivedusinga finite differencemethod.The time and space
c)(ll)oO - Oo
Oo oT intervals were chosen so that the Courant number was less
• (•Do 0 than 1 and the local Peclet number was less than 2. The time
0 0-
0
0
factor for the implicit finite differenceequationwas 0.6.
0 5 10 The velocityformula
Days

v(x)
=- • 0-•
=20z'ho r•
2 -• 1 r• '
o-4o M-1
r,D -5 o
wasusedin cu•e fitting. In theseequationsh o is the aquifer
(9)

-6

-7
0 5
, i , t
10
thickness,x is the coordinatealong a distancer from the
pumpingwell, r is the distancebe•een a well and a point
whereveloci• is to be calculated,and 0 is the aquiferporosi•.
Days In dehving(9) the aquiferhasbeen treatedas homogeneous
Figure 4. Colloid breakthroughcurvesfor first injectionfit- and isotropicso that 0 and K do not va• with x and aquifer
ted to the constantdispersivitymodel (attachment experi- depth. Equation (9) is exactlythe same as that when h is
ment). Circlesare data, solidlinesare simulations. definedas potential[Javandelet al., 1984].Equation(9) was
developedfrom fitting a Strackpotentialmodel [Strack,1985;
de Marsily,1986] to the head data from the obse•ation wells
andpiezometers[Li, 1993].We alsoevaluateduseof a delayed
responseof water table model [Neuman,1975] and a Theis
attachment rate constant for slow reaction sites, k 2 is the model [e.g.,Freezeand Che•, 1979],but neither gavea good
detachmentrate constantfor slowreactionsites,Pt, is aquifer fit to drawdowndata [Li, 1993]. Flow rates for an individual
bulk density,0 is aquifer porosity,and aa is the pore-scale level were varied to obtain the best fit. Dispersivi• estimates
dispersivity.
The concentrationof siteson the soil for viral or (0.1-0.6 m at M•) were derivedfrom fitting electricalcon-
microsphereattachmentis assumedto be largerelativeto the ductivi• breakthroughcu•es [Li, 1993].
concentrationof the virusesand microspheresand therefore Fitting was conductedonly on the completebreakthrough
doesnot appearin (2) and (3). cu•es, with resultsfor the first experimentshownon Figure4.
Equations(1)-(4) were used to describethe attachment Fitted chemicalparametersfor both attachmentand detach-
experiment.The samemodelbut with differentparameterswas ment experimentsare givenin Table 3. The simulatedcolloid
usedto describethe detachmentexperiment,that is, after the concentrationsin soil at day 10 of the attachmentexperiment
chemical disturbance.This was based on three assumptions: were used as input for the simulationof detachmentexperi-
(1) strongcolloiddetachmentstartedat a pointonlyafter the ment. The fitting results of the detachmentbreakthrough
pulseof high-pH water had arrivedat that point, (2) strong cu•es are shownin Figure 5.
colloid detachmentceasedafter the pulse had passedthat We alsopredictedcolloidbreakthroughcu•es at sampling
point, and (3) the high-pH water had residualeffectson the well ML5 usingthe parametersfrom M•. Sucha prediction
surfacesof soilparticlesandcolloidssuchthat reattachmentof dependedon successful simulationof both the attachmentand
colloidsat that point proceededat a rate differentfrom that detachmentexperimentsat M•. The predictionat ML5 for
previously.These assumptions required all rate constantsin the detachmentexperimentwas in agreementwith the ob-
the previousgoverningequations(2) and (3) to be treatedas se•ed timingof breakthrough(Figure 6). Both predictedand
functions of time:
obse•ed colloid concentrations were not zero. The predicted
peak timeswere near the obse•ed ones.The differencesbe-
rp={rpi2
Kp• ttx_<
<txor
t-< t>
tx tx+to
+ to (5) •een predictedandobse•ed peakconcentrations werewithin
a factorof 10. The modelpredictionfor the attachmentexper-
iment is not shown,sinceobse•ed breakthroughcu•es were
k•= {ki2
k• tx-<
t<txor
t _<t>
tx tx+to
+ to (6) incomplete.
 BALES ET AL.: BACTERIOPHAGE TRANSPORT 645

Table 3. Fitted Equilibriumand Rate Constants

ML k• k•2 k2 k22
Colloid Location Experiment day- day-• day-• day-• k3 k32
PRD-1 4-6 att. 10.7 ß.. 0.0003 ..- 2.05 ..'
PRD-1 4-6 det. 10.7 6.0 0.0003 15.0 0.25 0.25
PRD-1 4-8 att. 6.1 ß.' 0.0004 ... 2.12 ...
PRD-1 4-8 dct. 10.7 3.0 0.0050 17.0 1.05 1.05
M-1 4-6 att. 10.0 ß.. 0.0050 ... 2.35 ...
M-1 4-6 dct. 10.7 6.0 0.0005 8.0 0.25 0.25
Microsphere 4-6 att. 7.0 ß.. 0.0005 ... 0.05 ..-
Microsphere 4-6 dct. 7.0 3.0 0.0005 0.006 1.55 1.55
Microsphere 4-8 att. 11.2 ß.. 0.0070 ... 0.35 ...
Microsphere 4-8 det. 10.5 2.07 0.0080 1.0 1.25 1.25

Here att., attachment; det., detachment.

Discussion peak for conservativesolute,whereasthe peak microsphere

concentrationlaggedthe solutepeak. Concentrationsof both
At bothwellsthe microspheres were retardedlessthanwere colloids were attenuated. Colloid attenuation over the first
the two phages.The largestmicrosphereconcentrationvalues
observedat MIA were delayed0.7 daysrelativeto the electric
meterof transportwasa factorof 104-107for PRD-1 and
microspheres, respectively,a level that shouldbe indicativeof
conductivity,versusa delay of up to severaldaysfor PRD-1
transportof thesecolloidsover longertimes.Attenuationover
and M-1. At ML5, the largestmicrosphereconcentrationoc-
the next2 m wasapproximatelya factorof 10.Theseresultsare
curredbefore electricconductivitypeaksat levelsML5-4 and
consistentwith our previousnatural-gradienttransportexper-
ML5-6, and only 0.33 daysbehind at level ML5-8. Harveyet al.
iment at Cape Cod in whichmostof the attenuationoccurred
[1989] observeda similar range of behaviorin their natural-
betweenthe injection and first observationwells [Baleset al.,
gradient studyof microsphereand bacteria transport.In one
1995].
casethe peak bacteriaconcentrationwas observedbefore the
After the injectionof the high-pH water the dimensionless
PRD-1, M-l, and microspherepeak concentrations were 0.74,
ML4-6 ML4-8 0.15, and 0.003at samplingwell MIA and 0.002,0.002,and 3 x
! I , I 10-s at ML5, respectively.
Theseobservations
showed
that(1)
PRD-1 PRD-1
attachedphagesdetachedat high ratesand reenteredthe mov-
o-1
ing groundwater,(2) PRD-1 detachedmore readily than did
M-1 or the microspheres, and (3) phagesthat reenteredthe
movinggroundwaterreattachedto porousmedia as they were
o( transported downstream.Becausethe colloids observedat
MIA and ML5 in the secondexperimentwere from various
points between the injection well and samplingpoints, we
[d) I , I , I
could not accuratelyassessretardation in the secondexperi-
ment.

The combinedkinetic-equilibrium two-sitemodelwasfound

to be adequateto simulatecolloid attachmentand transport
processes (Figure4). The k • mainlyaffectsthe peakvalueof a
breakthroughcurve.The k 2 mainly influencesthe level of the
tail of the breakthrough.The ratio k•/k2 controlsthe slopeof
both the risingconcentrationand droppinglimbs of a break-
throughcurve.The k 3 affectsthe peakpositionfor a givenflow
rate and the width of the breakthroughcurvesas well. The
-7 ii , I , I , I i -7 ß ' , i , i
10 1'2 14 16 attachmentrate constantsof PRD-1, M-l, and microspheres
Days

-1
M-1
a. 5-6, PRD-1 b. 5-6, M-1 c. 5-8, PRD-1
o-1 -2
o
( o

o (•/•(
o•o o G

10 •'2 ' 14 16
0 I
I I
-7 , i ,

Days 10 12 14 16 10 1'2' 1'4,i16, 10 ' 1'2' 1"4' 1'6

Days Days Days
Figure 5. Colloid breakthroughcurvesfor secondinjection
fitted to the constantdispersivitymodel (detachmentexperi- Figure 6. Comparisonbetweenobservedand predictedcol-
ment). Circlesare data, solidlinesare simulations. loid breakthroughcurvesfor detachmentexperimentat ML5.
646 BALES ET AL.: BACTERIOPHAGE TRANSPORT

were 10,000-100,000timesthe corresponding detachmentrate effectof the pulseof high-pH water, that is, changingthe soil
constants,showingthat attachmentdominated(Table 3). and colloid surfacechemistry.
We previouslyreported a k• value for PRD-1 removal in During the attachmentand transportexperimentthe attach-
laboratorycolumnstudieswith soil from the site of about 100 ment of colloidswas fast and detachmentwasvery slowcom-
day-• [Kinoshita et al., 1993],10 timesthevalueestimated in paredto advection.Usingthe conductivitypeakinformationin
the currentwork. Velocity andpH in lab studieswere the same Table 1, the averagevelocity(over the distancefrom the in-
as in the current work, so this differenceshowsdirectly that jectionwell to the ML4 sampling well) was1.9 m d-•. The
phage removal in the field was only 10% as efficientas with timescaleof colloidattachmentrelativeto advection(u/k•L)
repackedsoil columns.Some differencemay be attributed to was 0.095 for PRD-1, 0.074 for M-l, and 0.088 for micro-
spatialheterogeneityin soil chemicalproperties.That is, the spheres,where L is the distancebetweenwells. The relative
soil used in the soil columns was from near the site of the time scalefor detachment(u/k2L) was2200 for PRD-1, 1500
experiment,but may not be representativeof the area of the for M-l, and 800 for microspheres.
experiment. Some if not most of the differencecan also be During the detachmentexperimentthe detachmentof col-
attributed to the use of repackedsoil versusphysicallyheter- loidswasfastand attachmentkept aboutthe samerate asthose
ogeneousfield media. Mean arrival times for the conservative in the attachmentexperiment.Using the conductivitypeak
tracerpeakwere relativelyconsistentin our experiment(ML4, information
in Table2, theaverage
velocity
was0.4m d-•. The
Table 1) but variedby _+35%in a previousexperimentat the timescale(u/k22L) was0.01for PRD-1, 0.02for M-l, and 13.2
site [Mackayet al., 1994]. for microspheres.These detachmenttimescaleswere much
Similarly,Mas-Pla et al. [1992] reportedthat the estimated smallerthan thosein the attachmentexperiment,varyingfrom
porosity value from one-dimensionalanalysisof a forced- 0.0004% for PRD-1, 0.013% for M-l, to 1.7% for micro-
gradient tracer experimentis physicallyunreasonable.They spheres.
attributedthe discrepancyto the fact that the one-dimensional
flow and transportdoes not capture three-dimensionalflow Conclusions
and transportphenomenain the field. As a result,the param-
eter valuesestimatedfrom the one-dimensionalanalysislikely As shownby our modelingof the field experimentof bio-
includeeffectsneglectedby the simplifiedanalysis. colloid transport, a one-dimensionaltransport model with
first-orderkinetic plus an equilibriummass-transfer
term was
Application of our transport model to the secondexperi-
adequateto describethe biocolloidmass-transferprocesses
ment,wherethe colloiddetachmentrate washigherthan in the
betweenthe soilandwaterphasesof the aquifer.The transport
first experiment,required consideringthe length of time the
of virusesand bacteria from sourcessuchas septictanks or
chemicaldisturbancehad an effect at any given point. Fast
sewagelines can therefore be simulatedwith the advection-
colloid detachment occurred when the chemical disturbance, a
dispersionequation coupledwith the simple mass transfer
shortdurationof high-pH water, passeda point. The strength
equations.However, our estimatedcoefficientsfor virus at-
of a disturbanceand the equilibrium and rate constantsde-
tachmentto soil particles,and thus log removal per length,
pendedon thep H value.Groundwaterp H at a givensampling
were only one tenth those found in lab studies.Since our
point varied up and downduringthe detachmentexperiment;
one-dimensionalmodel assumedhomogeneityand neglects
thusp H valuesat ML5 were about the sameas thoseat ML4
the effectof the partiallypenetratingwell (5-m screeninter-
(Figure 3). The equilibriumand first-orderrate constants
were
val), our model greatly simplified the tortuous three-
assumedto vary in responseto changesin p H, as shownby
dimensional flowregimein the aquifer.Thussomediscrepancy
(5)-(7). betweenthe parametervaluesof laboratoryexperimentsand
Figure 5 showsthat the combinedequilibrium and kinetic those of field tests is expected[Yeh, 1992; McCarthy et at.,
model can alsosimulatethe generalpattern of the detachment 1996]. This discrepancyhighlightsthe large uncertaintythat
and transport processes.In addition to the roles discussed will ensue using a one-dimensionalmodel with laboratory-
above, k•l, the first-order attachmentrate constantfor the determinedparameters.To properly interpret the chemical
times before and after the action of chemical disturbance at a
tracer data under field conditions, three-dimensional site char-
givenspatialpoint, affectsthe concentrationlevel and timingof
acterizationof aquiferheterogeneityand modeling[Yehet at.,
the beginningpart of a breakthroughcurve;k22, the first-order
1995a,b] may be necessary.
detachmentrate constantduring the action of the chemical In spite of possibleerrors due to the one-dimensionalanal-
disturbance,mainly controlsthe peak values.The PRD-1 de- ysis, we clearly showed that colloid attachment to the soil
tachment rate constantduring the chemicaldisturbancewas matrix followedby slow (relative to advectivetimescale)re-
106timesthatin the attachment experiment andof the same leaseof retainedcolloidswasmore importantthan wasrevers-
order asbut higherthan the attachmentrate constant.The rate ible "equilibrium"adsorption,becauseadsorptioncharacter-
constantsfor M-1 phageindicatedthe samerelationship.For ized by fastuptakeand slowreleasecontrolledthe final colloid
microspheres, the detachmentrate constantwas10-1000 times concentrationlevelsin both soiland groundwater.Equilibrium
that in the attachmentexperimentbut did not exceed the adsorptionon the timescaleof the experimentcontributedonly
attachment rate constant. Microsphere detachment was to a slight retardation of colloids and had a minor effect on
weakerascomparedto the two phages.The microsphereequi- concentration.
librium adsorptionconstant(k3) for the detachmentexperi- The pulseof high-pH water wasintroducedat the injection
ment was10-100 timesthat in the attachmentexperiment;the well as a chemical disturbance traveled downstream with the
phage equilibrium adsorptionconstantdecreasedby about groundwater. The actionof thischemicaldisturbance (causing
90%, showingagain that microspheresbehaved differently detachmentin our case)at a givenpoint downstreamstarted
from the phage.The lower attachmentrate constants k 1• (Ta- when it arrived at that point and ended when it passedby.
ble 3) during the detachmentexperimentshowsthe residual When the major actionwasto causedetachmentof previously
 BALES ET AL.: BACTERIOPHAGE
attachedcolloids,asin our case,further downstreamtransport
of colloids was facilitated. The rate of release of retained
colloids
caused
bya chemical
disturbance
was100to 106times
that of the unperturbedrelease.The detachmentand further Gerba, C. P., M. V. Yates, and S. R. Yates, Quantitation of factors
downstreamtransportof colloidscan be simulatedwith the controllingviral and bacterial transportin the subsurface,in Mod-
same type of model used for the attachmentand transport elingtheEnvironmentalFate of Microorganisms,
process,providedthe p H dependenceof the rate constantsis pp. 77-88, Am. Soc.for Microbiol., Washington,D.C., 1991.
included.
Finally, prediction of bulk behavior of virus transport is
possiblewith appropriateestimatesof physicochemical param-
eters, primarily attachmentand detachmentrate coefficients;
simplyknowinga "retardationfactor" is not sufficient.Exact
predictionof transport,suchas the number of virusesoccur-
ring at a certainpoint downstreamfrom a source,may not be
feasibleconsideringthe heterogeneityof aquifer in its physi-
cochemicalqualities. But order-of-magnitudeestimatesmay
be sufficientfor engineeringsolutionsto biocolloidtransport
problems.Resultsof recent controlledfield experimentsand
detailed three-dimensional modeling efforts by Yeh et al.
[1995a,b] supportthis conclusion.
Acknowledgments. The project describedwas supportedin part by
funding from the U.S. EnvironmentalProtection Agency's(EPA)
Robert S. Kerr Environmental Research Laboratory in Ada, Okla-
homa, throughgrant CR-818113, with additionalsupportfrom grant
P42ES04940 from the National Institute of Environmental Health
Sciences,NIH. This paper doesnot necessarily
reflect the viewsof the
NIH or EPA. We also acknowledgethe generous support of D.
Mackay and J. Cherry for givingus use of the site and G. Bianchi-
Mosquera for cooperationin carryingout the experiment.R. Brice
assistedin manuscriptpreparation.
References
Adams, M. H., Bacteriophages, Interscience,New York, 1959.
Bales, R. C., S. R. Hinkle, T. W. Kroeger, K. Stocking, and C. P.
Gerba, Bacteriophageadsorptionduring transport through porous
media: chemicalperturbationsand reversibility,Environ.Sci. Tech-
nol., 25, 2088-2095, 1991.
Bales, R. C., S. Li, K. Maguire, M. Yahya, and C. P. Gerba, MS-2
phageand poliovirustransportthroughporousmedia: sorbenthy-
drophobicityand chemicalperturbations,WaterResour.Res., 29,
957-963, 1993.
Bales, R. C., S. Li, K. M. Maguire, M. T. Yahya, C. P. Gerba, and
R. W. Harvey, Virus and bacteriatransportin a sandyaquifer, Cape
Cod, MA, Ground Water,33(4), 653-661, 1995.
Bamford, D. H., L. Rouhiainen, K. Takkinen, and H. S6derlund,
Comparisonof the lipid-containingbacteriophagesPRD1, PR3,
PR4, PR5 and L17, J. Gen. Virology,57, 365-373, 1981.
1996.
Bamford, D. H., J. Caldentey,and J. K. H. Bamford, Bacteriophage
PRDi: A broad hostrangeDsDNA tectiviruswith an internal mem- McCaulou, D. R., R. C. Bales, and R. G. Arnold, Effect of tempera-
brane, Adv. Virus Res., 45, 281-319, 1995. ture-controlledmotility on transportof bacteria and microspheres
Bear, J., Hydraulicsof Groundwater,721 p., McGraw-Hill, New York, through saturatedsediment, Water Resour.Res., 3•(2), 271-280,
1979.
Cameron, D., and A. Klute, Convective-dispersive
solute transport
with combinedequilibrium and kinetic adsorptionmodel, Water
Resour. Res., 13, 183-188, 1977.
Corapcioglu,M. Y., and A. Haridas, Transportand fate of microor-
ganismsin porousmedia: a theoreticalinvestigation,
J. Hydrol.,72,
149-169, 1984.
Corapcioglu,M. Y., and A. Haridas, Microbial transportin soilsand
groundwater:A numericalmodel,Adv. WaterResour.,8, 188-200,
1985.
de Marsily,G., QuantitativeHydrogeology,
Academic,SanDiego, Calif.,
1986.
Frankel-Conrat,H., P. C. Kimball, and J. A. Levy, Virology,Prentice
Hall, EnglewoodCliffs,N.J., 1988.
Freeze, R., and J. Cherry, Groundwater,Prentice Hall, Englewood
Cliffs, N.J., 1979.
Funderburg,S. W., B. E. Moore, B. P. Sagik,and C. A. Sorber,Viral
TRANSPORT 647
transport through soil columnsunder conditionsof saturatedflow,
Water Res., 15, 703-711, 1981.
Gerba, C. P., Applied and theoretical aspectsof virus adsorptionto
surfaces,Adv. Appl. Microbiol.,30, 133-168, 1984.
editedby C. J. Hurst,
Grosser, P. W., A one-dimensional mathematical model of virus trans-
port, in Proceedingsof the Second International Conferenceon
GroundwaterQualityResearch,pp. 105-107, Univ. Ctr. for Water
Res., Okla. State Univ., Stillwater, 1985.
Harvey, R., Parametersinvolvedin modelingmovementof bacteria in
groundwater,in ModelingtheEnvironmentalFate of Microorganisms,
edited by C. J. Hurst, Am. Soc.for Microbiol., Washington,D.C.,
1991.
Harvey, R. W., and S. P. Garabedian,Use of colloidfiltration theory in
modelingmovementof bacteriathrougha contaminatedsandyaqui-
fer, Environ. Sci. Technol., 25, 178-185, 1991.
Harvey, R., L. H. George, R. L. Smith, and D. R. LeBlanc, Transport
of microspheresand indigenousbacteria through a sandy aquifer:
Resultsof natural and forced-gradienttracer experiments,Environ.
Sci. Technol., 23, 51-56, 1989.
Hobbie, J. E., R. J. Daley, and S. Jasper,Use of nucleporefilters for
countingbacteriaby fluorescencemicroscopy, Appl. Environ.Micro-
biol., 33, 1225-1228, 1977.
Javandel,I., C. Doughty,and C. F. Tsang(Eds.), GroundwaterTrans-
port:Handbookof MathematicalModels,Amer. Geophys.Union, Wa-
ter Res.Monogr.,vol. 10, AGU, Washington,D.C., 1984.
Kinoshita,T., R. C. Bales,M. T. Yahya, and C. P. Gerba, Effect ofp H
on bacteriophagetransportthroughsandysoils,J. Contam.Hydrol.,
15, 55-70, 1993.
Li, S., Modeling biocolloidtransportin saturatedporousmedia, Ph.D.
thesis, Univ. of Ariz., Tucson, 1993.
MacFarlane, D. S., J. A. Cherry, R. W. Gillham, and E. A. Sudicky,
Migration of contaminantsin groundwaterat a land fill: A case
study, 1, Groundwaterflow and plume delineation,J. Hydrol., 63,
1-29, 1983.
Mackay, D. M., G. Bianchi-Mosquera,A. A. Kopania, H. Kianjah, and
K. W. Thorbjarnarson, A forced-gradient experiment on solute
transportin the Borden aquifer, 1, Experimentalmethodsand mo-
ment analysesof results,WaterResour.Res.,30(2), 369-383, 1994.
Mackay, D. M., D. L. Freyberg,P. V. Roberts, and J. A. Cherry, A
naturalgradientexperimenton solutetransportin a sandaquifer, 1,
Approachand overviewof plume movement,WaterResour.Res.,22,
2017-2029, 1986.
Mas-Pla,J., T.-C. J. Yeh, J. F. McCarthy, and T. M. Williams, Numer-
icalsimulationof a two-welltracerexperiment,GroundWater,30(6),
958-964, 1992.
Matthes, G., A. Pekdeger,and J. Schroeter,Persistenceand transport
of bacteria and virusesin groundwater--a conceptualevaluation,
J. Comtam.Hydrol., 2, 171-188, 1988.
McCarthy, J. F., B. Gu, L. Liang, J. Mas-Pla, T. M. Williams, and
T.-C. J. Yeh, Field tracer testson the mobility of natural organic
matter in a sandyaquifer, WaterResour.Res., 32(5), 1223-1238,
1995.
Neuman, S. P., Analysisof pumpingtest data from anisotropicuncon-
fined aquifersconsideringdelayedgravity response,WaterResour.
Res., 11, 329-342, 1975.
Olsen,R. H., J. Siak,and R. Gray, Characteristics
of PRD1, a plasmid-
dependentbroad host range DNA bacteriophage,J. Virology,14,
689-699, 1974.
Strack, O. D. L., GroundwaterMechanics,Prentice Hall, Englewood
Cliffs, N.J., 1985.
Sudicky,E. A., A natural gradient experimenton solutetransportin a
sandaquifer:Spatialvariabilityof hydraulicconductivityand its role
in the dispersionprocess,WaterResour.Res., 22(13), 2069-2082,
1986.
Vilker, V. L., An adsorptionmodel for prediction of virus break-
throughfrom fixedbeds,in Stateof Knowledgein Land Treatmentof
Wastewater, vol. 2, edited by H. L. McKim, pp. 381-388, U.S. Army
Cold Reg. Res. and Eng. Lab., Hanover, N.H., 1978.
648 BALES ET AL.: BACTERIOPHAGE TRANSPORT

Vilker, V. L., and W. D. Burge, Adsorptionmasstransfermodel for sandyaquifer under forced-gradientconditions,WaterResour.Res.,

virus transportin soils,WaterRes., 14, 783-790, 1980. 31(9), 2141-2157,1995b.
Yahya, M., L. Galsiomes,C. P. Gerba, and R. C. Bales, Survivalof
bacteriophagesMS-2 and PRD-1 in groundwater, WaterSci. Tech- R. C. Balesand T.-C. J. Yeh, Department of Hydrologyand Water
nol., 27, (3-4), 409-412, 1993. Resources,University of Arizona, Box 210011, Tucson, AZ 85721.
Yates, M. V., S. R. Yates, J. Wagner, and C. P. Gerba, Modelingvirus (e-mail:roger@hwr.arizona.edu)
survivaland transportin the subsurface, J. Contam.Hydrol.,1,329- C. P. Gerba, Department of Soil, Water, and EnvironmentalSci-
345, 1987. ence, University of Arizona, Tucson,AZ 85721.
Yeh, T.-C. J., Stochasticmodeling of groundwaterflow and solute M. E. Lenczewski,Amway Corp., 7575 Fulton St. E 50-2D, Ada, MI
transportin aquifers,J. Hydrol.Process.,6, 369-395, 1992. 49355-0001.
Yeh, T.-C. J., J. Mas-Pla, J. F. McCarthy, and T. M. Williams, Mod- S. Li, Arizona Department of EnvironmentalQuality, 3033 North
eling of natural organicmatter transportprocesses in groundwater, Central Avenue, Phoenix, AZ 85012.
Environ.HealthPerspect., 108 (suppl.I), 41-46, 1995a.
Yeh, T.-C. J., J. Mas-Pla, T. M. Williams, and J. F. McCarthy, Obser- (ReceivedApril 22, 1996;revisedDecember31, 1996;
vation and three-dimensionalsimulationof chloride plume in a acceptedDecember31, 1996.)