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Ledger, M.L., Stock, F., Schwaiger, H., Knipping, M., Brückner, H., Ladstätter, S., Mitchell, P.D. (2018) Intestinal parasites from public and private latrines and the harbor canal in Roman period Ephesus, Turkey (1st c. BCE to 6th c. CE). Journal of Archaeological Science Reports 21: 289-97.
MarissaL.Ledger1,FriederikeStock2,5,HelmutSchwaiger3,MariaKnipping4,Helmut
Brückner2,SabineLadstätter3,PiersD.Mitchell1*
1DepartmentofArchaeology,UniversityofCambridge,TheHenryWellcomeBuilding,FitzwilliamStreet,
CambridgeCB21QH,UK2 InstituteofGeography,UniversityofCologne,ZülpicherStr.45,50674Köln,Germany3AustrianArchaeologicalInstitute,AustrianAcademyofScience,LesFranz-Klein-Gasse1,1190Vienna,Austria4InstituteofBotany,UniversityofHohenheim,SchloßHohenheim1,70599Stuttgart,Germany5FederalInstituteofHydrology,AmMainzerTor1,56068Koblenz,Germany*correspondingauthor
E-mailaddresses:
[email protected](M.L.Ledger),
[email protected](F.Stock),
[email protected](H.Schwaiger),
[email protected](M.Knipping),
[email protected](H.Brückner),
[email protected](S.Ladstätter),
[email protected](P.D.Mitchell)
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Abstract
ToimproveourknowledgeoftheparasitespeciesaffectingtheinhabitantsofRomanperiod
AsiaMinor,weanalysedfaecalmaterialfromEphesus,Turkey.Mineralisedmaterialfrom
thedrainfromaprivatehouselatrine(3rdc.CE),sedimentsamplesfromthesewerdrainof
apubliccommunallatrine(6thc.CE),andsedimentfromtheharbourcanal(ca.1stc.BCEto
ca.6thc.CE)werestudiedforthepresenceofintestinalparasites.Sampleswereviewedby
lightmicroscopyforhelmintheggs,andcommercialenzyme-linkedimmunosorbentassay
(ELISA)kitswereusedtotestforprotozoalparasitesthatcausedysentery.Eggsof
roundwormwerefoundinthepubliclatrine,whipworminthehouselatrine,andboth
whipwormandroundwormintheharbourcanal.Sequentialsamplingoftheharbourcore
suggeststhatwhipwormwasbyfarthemostcommonparasitethroughouttheRoman
period,andtherewasnoclearevidenceforchangeinparasitespeciesoverthecenturies.
Whipwormandroundwormarebothspreadbythecontaminationoffoodanddrinkby
humanfaeces.DespitethelargenumberoftravellerstoEphesus,asthecapitalofits
provinceandamajorportcityintheRomanEmpire,therewasasurprisinglackofdiversity
inparasitespeciesfound.Thisisespeciallyapparentwhenweconsiderthattenspeciesof
intestinalparasitehavebeenfoundacrosstheRomanEmpire.ThisisthefirstRomansiteto
bedirectlyassessedfordifferencesbetweeninfectioninindividualsusingprivatelatrines,
publiclatrines,andmixedtowneffluent(intheharbour)atthesamesite.
Keywords:Palaeoparasitology;Ephesus;Romanempire;Helminths;Roundworm;
Whipworm
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1.Introduction
Thereisafairrangeofarchaeologicalandhistoricalevidenceforinfectiousdisease
intheRomanperiod.Particularfocushasbeenplacedonmajorpandemicsthatareknown
tohavekilledlargeportionsofthepopulation,includingtheAntoninePlague,thePlagueof
Cyprian,andthePlagueofJustinian(RetiefandCilliers,2000;Little,2007;Wagneretal.,
2014;Harper,2015;McCormick,2015).Throughthestudyofskeletalremainstherehave
alsobeenaccountsofotherinfectionsthatcanproducelesionsontheskeletonincluding
tuberculosis,treponemaldisease,andleprosy(RobertsandCox,2003;Cancietal.,2005;
Lewis,2011;Minozzietal.,2012;Rissechetal.,2013;Mülleretal.,2014;Rubinietal.,
2014).MostoftheevidencewehaveforinfectionintheRomanperiodcomesfromstudies
focusingonbacteria.However,evidenceforparasiticinfectionintheRomanperiodis
steadilygrowing.
Parasitesareoneofthefourmaincategoriesofpathogens,alongsidebacteria,
viruses,andfungi.Thetwomaintypesofparasitesareendoparasites,thosethatliveinside
thebody,andectoparasites,whichliveoutsidethebody.Endoparasites,suchasintestinal
wormsandsinglecelledprotozoa,areofparticularinteresttopalaeoparasitologists
studyingdiseaseinpastpopulationsbecausetheyofferinsightsintohumanmigration,diet,
sanitation,andinteractionwithanimals(Reinhard,1992;LeBaillyandAraújo,2016;Araújo
etal.,2008;Mitchell,2013).ArchaeologicalevidenceforparasitesintheRomanperiodhas
suggestedthatintestinalparasiteswerecommoncontributorstodiseaseinRoman
populationsinmanyareasoftheEmpire(Mitchell,2017).Thisisdespitetheadventof
centralizedsanitationmeasuresandwidespreaduseoflatrinesandsewersduringthe
Romanperiod,whicharemeasuresthataresupportedbymodernepidemiologicalstudies
todecreasetheincidenceofparasiticinfection(Feachemetal.,1983;Ziegelbaueretal.,
2012;Speichetal.,2016).
ThemaximumexpansionoftheRomanEmpirewasreachedinthe2ndcenturyCE,
whenitsbordersextendeduptonorthernBritaininthenorth,theAtlanticOceaninthe
west,northernAfricainthesouth,andtheEuphratesriverintheeast.Parasiticevidence
fromtheRomanprovinceofAsia(coveringmostofmoderndaywesternTurkey),isslim
comparedtootherprovinces,especiallythoseinnorthernEurope.Infact,onlyoneother
archaeologicalsitefromTurkeyhaseverbeenstudiedforparasiticinfection,theRoman
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periodsiteofSagalassos(Williamsetal.,2017).Thisstudyaimstocontributetoevidenceof
parasiticinfectioninRomanperiodAsiaMinorthroughanalysisofsedimentsamplesfroma
publicmulti-seatlatrine,aprivatehouselatrine,andasedimentcorefromtheharbour
canalatEphesus.Usingthesedifferentarchives,wehopedtodetectvariationsinparasite
infectionamongthepopulationsofthecity.
2.RomanperiodEphesus
ThecityofEphesus(Fig.1)hasbeenwellstudiedbothfromahistoricalandarchaeological
perspectiveduetoitsimpressiveruinsandreligioushistory.Itisthesiteofthefamed
TempleofArtemisandthelibraryofCelsus.Ephesushasalonghistoryofoccupation
beginninginthe7thmillenniumBCE.TheHellenisticcitywasfoundedbyoneofAlexander's
successor,Lysimachos,in296BCE,andbecamepartoftheRomanRepublicin133BCE
(Ladstätteretal.,2016).UnderthereignofAugustus,Ephesuswasdeclaredthecapitalof
theprovinceofAsia,inplaceoftheformercapitalPergamum,afterwhichthecitygrew
extensivelythroughtheImperialperiod(Scherrer,2001).Itwassituatedalongroads
connectingtheAegeancoasttotheinteriorofAsiaMinoraswellasthePersianRoyalRoad
andtraderoutesthroughtheMeandervalley(Ladstätter,2018).Itwasalsowellconnected
totherestoftheMediterraneanregionbyitsharbour.Historicalrecordsgivefurther
evidenceformanyhigh-statusvisitorswhocametoEphesusforpoliticalreasons.Asthe
seatoftheproconsulinAsiaandacentreforearlyChristiancommunities,itwasacentrefor
commerce,politics,andscholarship,particularlyinmedicine.
Archaeologicalexcavationsbeganatthesitein1863byJ.T.Woodinhissearchfor
theTempleofArtemis.Since1895excavationshavebeencontinuedbytheAustrian
ArchaeologicalInstitute,uncoveringpublicandprivatespacesoftheLateAntique/Byzantine
cityandharbourarea(Ladstätteretal.,2016).Sedimentsamplescollectedduringrecent
excavationswereusedtolookforevidenceofparasiticinfectioninthepopulationofthe
city(Fig.2).
3.Materialsandmethods
Thefirstsetofsedimentsampleswerecollectedfromthedrainofapubliclatrine
associatedwiththeadjacentVariusbathcomplex,alsocalledtheBathsofScholastikia
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(Jansen,2006).Thisbathcomplexwaslocatedalongamajorpublicstreetthroughthe
ancientcity.Thepubliclatrinewasavailableforusebyanyonevisitingthebaths.Thismeans
itwouldhaveattractednotonlylocalsbutalsothosefromotherprovinceswithinthe
EmpirewhowerevisitingEphesusfortrade,politics,scholarship,orpassingthroughon
theirwaytoothercities.Sedimentsamplesweretakenatfourdepthsfromthesurface(0–
20cm,40–60cm,90–110cm,andat150cm)inthedrainofthepubliclatrine,after
modernsedimenthadbeenremoved(Fig.3).Allsamplesdatetothe6thcenturyCE,the
latestperiodofuseofthelatrine.
Thesecondsetofsampleswerecollectedfromaprivatelatrine(RoomNo.34/34a)
ofoneofthenearbyTerracehouses,datedfromtheAugustanperiodtothe3rdcenturyCE
(Ladstätter,2013;Ployer,2016).ArchaeologicalexcavationoftheTerracehousesatEphesus
hasshownthattheywerequiteopulentandwouldhavebeeninhabitedbyhigherclass
individualswhowereabletoaffordtheluxuryofaprivatelatrine.Mineralisedmaterial
adherenttothesidesofthesewerconnectedtothelatrinefromTerraceHouse2(unit7)
wasscrapedoffandusedforanalysis(Fig.4b).Theseatingcomponentofthelatrinewas
destroyedinthe3rdcenturyCE(Fig.4a).
Thethirdsetofsampleswerecollectedfromasedimentcore(Eph244)fromthe
harbourcanal,whichconnectedtheRomanharbourbasinofEphesuswiththeopenseaof
theAegean(Fig.2).Thecanalwasbuilttomaintainaccesstothecityforshipping,asthe
harbourtendedtosiltupwithsedimentfromtheCaystros(modernname:KüçükMenders)
delta.Weanalysedsequentialsectionsofthesedimentcoredatingfromthe1stcenturyBCE
tothe6thcenturyCE.Weanticipatedthatsewersanddrainsfromthecitywouldopeninto
thewaterfront,thuscarryingtheeggsofintestinalparasitesintotheharbour.Resultsofthe
sedimentcoreanalysisformicrofauna,pollen,spores,plantfragments,andshellswere
publishedelsewhere,andmentionwasmadeoftheparasitespeciespresentinthe
sediment(Stocketal.,2016).Thedatesforallthesesampleshavebeendeterminedfroman
intensiveseriesofradiocarbondatesthathavebeenundertakenacrosstheexcavationat
Ephesus.
SampleswereanalysedinthePalaeoparasitologyLabattheUniversityofCambridge,
UK.A0.2gsubsamplewasfirstdisaggregated(madeintoaliquidsuspension)torelease
parasiteeggsfromothermaterialinthesample.Forthepubliclatrinesediment,0.5%
aqueoustrisodiumphosphatewasusedfordisaggregation.Afteraperiodof2hourswith
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periodicshakingofthetesttube,thesedimentwascompletelydisaggregated.Theresulting
suspensionwaspouredthroughastackofmicrosieveswithmeshsizesof300µm,160µm,
and20µmontopofacatchmentcontainer.Thetypicalrangeforthedimensionsofhuman
parasiticwormeggsisbetween22µm(width)and140µm(length)(Garcia,2016).
Therefore,anyparasiteeggsinthesamplewouldhavebeentrappedontopofthe20µm
sieve.Thismaterialwascollectedandcentrifugedat4,000rpmfor5minutes.The
supernatantwasremovedandtheremainingmaterialmixedwithglycerolandviewedat
400´magnificationonalightmicroscopeequippedwithadigitalcamera(Anastasiouand
Mitchell,2013).
ForthemineralisedmaterialfromtheprivatelatrineofTerraceHouse2,0.5%
trisodiumphosphatewasnotsufficienttobreakuptheconcretions.Dilutehydrochloricacid
(HCl)wasusedtobreakdowncalciumcarbonatestoreleaseparasiteeggsfromthe
concretion.HClwasaddeddropwisetothesampleuntilareactionoccurred,evidentby
gentlebubblingduetoproductionofcarbondioxide.Eachtimethereactionstopped,HCl
wasaddeddropwisetocontinuethereactionuntilthesamplewascompletely
disaggregated.Thesamplewascentrifugedandwashedwithtrisodiumphosphatefive
timestodiluteandwashoutanyremainingHCl.Itwasthenmixedwithglycerolandviewed
underthemicroscope.DisaggregationusingHClhasbeenevaluatedincomparisontoother
reagents,anditwasobservedthatHClcandecreasespeciesdiversityandshouldonlybe
usedwhenmaterialcannotbeprocessedusingtypicalnon-destructivesolutions(Dufour
andLeBailly,2013).
Eachsamplewasalsotestedforsingle-celledprotozoalparasitesthatcause
dysentery,includingEntamoebahistolytica,Giardiaduodenalis,andCryptosporidium
parvum,usingenzyme-linkedimmunosorbentassay(ELISA).ELISAisanimmunological
methodthattestsforthepresenceorabsenceofspecificorganismsusingaplatewithwells
containingimmobilizedantibodyspecifictothepathogeninquestion.Ifthepathogen,or
thespecificantigenbeingtargeted,ispresentitbindstotheantibodyinthewells.Another
antibodylinkedtoanenzymeisthenaddedwhichbindstothiscomplexandcausesavisible
colourchange,whichconfirmsthepresenceofthepathogen.CommercialELISAkitsfrom
TechLab©,designedandtestedonclinicalsamplesemployingamonoclonalantibody-
peroxidaseconjugatespecificforproteinsuniquelysecretedbytheseorganisms,wereused
foranalysis.Thesensitivityandspecificityforeachofthekitsisboth100%forGiardia
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duodenalis,96.9%sensitivityand100%specificityforEntamoebahistolytica,and97.7%
sensitivityand100%specificityforCryptosporidiumparvum(Sharpetal.2001).Foreach
sample,thedisaggregatedandsievedsedimentfromunderneaththe20µmsievewas
collectedandusedforthetests,astheprotozoalcystsaretypicallylessthan20µminsize
(Garcia,2016).TheELISAtestswerecarriedoutfollowingthemanufacturer’sprocedure.A
negativeandpositivecontrolwerepresentoneachELISAplate,andanELISAplatereader
wasusedtoquantifycolourchangesineachwell.
4.Results
Resultsfromthemicroscopyrevealedparasiteeggsinallsamplesstudied.Samples
fromthepubliclatrinecontainedeggsofroundworm(Ascarissp.).Theseeggswerefoundin
samplesfromallfourdepths,althoughahigherconcentrationofeggswerefoundinthe
middletwolayers,from40–60cmand90–110cm.Atotaloftwentyfertilizedroundworm
eggswerefoundinthesamplesfromthepubliclatrine(Fig.5).Twosubsampleswere
studiedfromthemiddletwolayerstoensurethatnospeciesweremissedonanalysis;this
gaveaconcentrationof17eggspergraminthelatrine.Thissuggeststhattheegg
concentrationwasnothigh.ThetypicalsizerangeofmodernAscarislumbricoides,the
speciesthatinfectshumans,is40–75µmlongand35–50µmwide(Garcia,2016).Theeggs
foundinthepubliclatrinefromEphesushadmeandimensionsof60.9µmlongand46.5µm
wide(Table1).Alloftheroundwormeggsfoundwerecompletelyorpartiallydecorticated,
havinglosttheiroutermammillatedcoat.Thisisquitecommonforeggsfoundfrom
archaeologicalcontexts,asthemammillatedcoatontheoutsideofthechitinouswallofthe
eggsismoresusceptibletodegradationbytaphonomicprocesses(Ráczetal.,2015).When
theeggsaredecorticated,theirdimensionswillbeafewµmsmallerthanwhenthe
mammillatedcoatremains.
Eggsofhumanwhipworm(Trichuristrichiura)werefoundinthemineralised
materialfromtheprivatelatrineatTerraceHouse2(Fig.6).Atotalof11eggswerefoundin
0.2gofmaterial,aconcentrationof55eggspergram.Themeanlengthofeggsfoundwas
50.1µmandthemeanwidthwas26.4µm(Table1).Thetypicalsizerangeofwhipworm
eggsis34–57µmlongwithoutpolarplugs,or49.5–65µmlongwithpolarplugs,and20–30
µmwide(Beer,1976;Garcia,2016).
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Samplesfromtheharbourcanalsedimentcorewerefoundtocontaintheeggsof
bothroundwormandwhipworm(Fig.7).Weanalysedsedimentdatedtoca.the1stcentury
BCEtoca.the6thcenturyCE,spanningtheentiretimeperiodoftheothersamplesstudied
inthecity.Ninesamplesweretakenatregularintervalsfromalongthecore.Whileboth
whipwormandroundwormeggswerefoundinthesediments,theywereinaratioof19:1
whipwormtoroundworm.WhipwormwasfoundineverysamplethroughouttheRoman
period.
Parasite SampleLocation Date
No.of Length(µm) Width(µm)eggs Range Mean σ Range Mean σ
Roundworm PublicLatrine 6thc.CE 20 54.6–66.4 60.9 3.4 40.3–51.7 46.5 2.8
WhipwormTerraceHouseLatrine
3rdc.CE 11 44.8–55.7 50.1 3.6 24.2–28.0 26.4 1.2
Table1.LocationofparasiteeggsfoundinlatrinesatEphesus,parasiteeggcounts,sizerange,meandimensions,andstandarddeviations(σ).
ELISAtestswerenegativeforallthreeprotozoa,Giardiaduodenalis,Entamoeba
histolytica,andCryptosporidiumparvum.Thepositiveandnegativecontrolshadthe
appropriatecolourchangeandabsorbancevaluestoconfirmthatthetestwasundertaken
properlyandtherewerenodesignerrorsintheplates.
5.Discussion
WehaveinvestigatedparasitismintheRomanperiodpopulationofEphesusby
analysingcomplementarysamplesfromacommunalpubliclatrine,aprivatehouselatrine,
andtherunofffromcitydrainsandsewersintotheharbour.Wefoundgoodevidencethat
whipwormandroundworminfectedsomemembersofthepopulation.
Roundworm(Ascarissp.)isoneofthemostcommonintestinalwormsbothinthe
pastandtoday.Itistransmittedviathefaecal-oralroutewheneggsofthewormpolluting
soilsandwaterareingested.Whenahumanisinfectedwithroundworm,theworm
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releasesapproximately240,000eggsperdayinthatindividual'sfaeces(WellerandNutman,
2014).Theseeggsthenmatureintheexternalenvironmentoveraperiodof2weeksat
whichpointtheybecomeinfective,anduponingestionbythesameoranotherindividual
theycandevelopintoadultwormsinthesmallintestine.Theadultroundwormis15-30cm
longandcanlivefor2years(Jourdanetal.,2017),absorbingnutrientsfromthegutbefore
theycanbeabsorbedforusebytheindividual.Thesymptomsexperiencedbyaninfected
individualvarybasedonage,comorbidities,healthstatus,andwormburden(thenumberof
wormspresentatonetime).Roundworminfectionmaybeasymptomatic,butinfectioncan
beparticularlydetrimentaltochildrenandthosewithahighwormburdenresultingin
anaemia,abdominalemergenciessuchasintestinalobstruction,pancreatitis,cholecystitis,
andliverabscess(Jourdanetal.,2017).Commonsymptomsincludeabdominaldiscomfort,
alteredbowelhabits,decreasedenergy,andweightloss.
Whipworm(Trichuristrichiura)isasoil-transmittedhelminth,likeroundworm.An
infectedindividualshedseggsintheirfaeces.Theseeggsmatureinthesoilandinfectnew
individualswhentheyareingested.Theadultwhipwormgrowstoaround3cmlonginthe
largeintestineandcolon(Garcia,2016).Thewormreleasesonaverage5,000eggsperday
andlivesforaperiodofupto5years(WellerandNutman,2014).Whipwormburrowsinto
themucosaoftheintestineandcancauseinflammationandbleeding;asaresult,anaemia
canbemoreseverewithwhipworminfectioncomparedtoroundworminfection(Jourdan
etal.,2017).Similartoroundworm,symptomsincludeabdominalpain,changeinbowel
habits,weightloss,andwithahigherwormburden,rectalprolapse.
Sanitationmeasuressuchaslatrineuseandhandwashinghavebeenshownto
significantlydecreasethetransmissionofwhipwormandroundworm(Zeigelbaueretal.,
2012).However,despitetherelativelylowconcentrationofeggsnoted,itappearsthat
latrinesdidnotfullyprotectthepopulationatEphesus.Thiscouldbeduetoanumberof
reasons.ItisunlikelythateveryoneinEphesuswouldhaveusedlatrines,andinhousesthat
didnothavetheirownprivatelatrinesfaecalmaterialwouldhavebeencollectedand
dumpedintoopensewers,streets,ordirectlyintowagonstobetakenaway(Taylor,2015).
Similarly,thosewithoutaccesstolatrineswhentheyneededthemmayhavedefecated
elsewhereinthecity,suchasinstreets,doorways,andbehindstatues(Scobie,1986).Inthis
environment,onecouldcomeintocontactwithhumanexcrementastheyventuredthrough
thecityonanynumberoferrands.Furthermore,althoughthereisevidenceforrunning
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waterinthelargepubliclatrineswhichwasavailableforhandwashingafteruseofthe
facilities,wedonotknowtowhatextenthandhygienewasemphasized.Forthose
individualswhorelievedthemselvesinthestreets,thepublicfountainsmayhavealso
servedthepurposeofaplacetorinsehands.Manypeoplelivinginlargecitieswouldhave
collecteddrinkingwaterfrompublicfountainsorbasinswhichcouldhavebeen
contaminatedbythesedirtyhands,andcontainersusedtocollectwater(Scheidel,2009).
Finally,humanfaeceswereknowntobeaneffectivefertilizerforcropsintheRoman
Empire.Therearenumerouswrittenrecordsandarchaeologicalevidenceattestingtothe
useofhumanfaecesasfertilizerafteritwascollectedfromRomancities(Farmer,1918;
White,1970;Baetenetal.,2012;Koloski-Ostrow,2015).Thisprocessoffersanidealroute
fortransmissionofintestinalwormswhencropscontaminatedwithinfectiveparasiteeggs
fromfreshhumanfaecesaretheneatenbyahighproportionofthepopulation.
EphesuswasamajortradingcenterinRomanAsiaMinor.Assuchitisinteresting
thatwedidnotfindanyevidenceforotherintestinalparasitespeciesthathavepreviously
beenidentifiedintheRomanEmpire.Besidesroundwormandwhipworm,thelancetliver
fluke(Dicrocoeliumdendriticum),Taeniatapeworms,fishtapeworm(Diphyllobothrium
spp.),Fasciolaliverfluke,Capillariaspp.,pinworm(Enterobiusvermicularis),andprotozoa
causingdysentery(GiardiaduodenalisandEntamoebahistolytica)haveallbeenfoundin
faecalsamplesstudiedfromRomanperiodsites(Rouffignac,1985;Horne,2002;Harter,
2003;LeBaillyandBouchet,2010;LeBaillyandBouchet,2013;Mowlavietal.,2014).There
wasalsothepotentialtofindtheeggsofschistosomesfromthosepeoplewhotravelledto
EphesusfromAfricaortheMiddleEast(AnastasiouandMitchell,2015).Currently,the
majorityofdatathatexistsforparasitesintheRomanEmpirecomefromsitesthathave
beenstudiedinnorthernEurope,especiallyFranceandBritain.Intheseareas,amore
diverserangeofspecieshasbeenfoundinarchaeologicalmaterialcomparedtothe
Mediterraneanregion.
ThelackofparasitespeciesdiversityatEphesusmakesusconsideroptionsforwhy
thismightbethecase.Onepossibilityisthatanimalmeatmayhavebeenwellcooked
beforeitwaseaten,killingparasitelarvaeinthemeatandsopreventinginfection.Historical
evidencesuggeststhateatingrawmeatwasunusualinhotterregionsoftheRomanEmpire
andthatcookingprocessesadoptedfromtheGreeksreliedonboilingmeatwhichcould
thenbefollowedbyroasting(Chandezon,2015).Thisextensivecookingprocesswould
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allowthemeattoreachtemperaturesthatwouldkillparasitesinporkorbeef,effectively
removingtheriskforTaeniatapeworms.ThisisincontrasttonorthernEuropeinthepast,
wheretherewasmoreofatraditionofeatingraw,pickled,driedorsaltedanimalproducts
(Mitchell,2017).AnotherpossibilityisthatthedietofthoseatEphesuswashighly
dependentuponfarmedfoodsratherthanwildfoodshuntedandcaughtfromthe
countryside.ThismayexplaintheabsenceofzoonoticparasitesatEphesusthatarespread
bytheconsumptionofthesewildanimals.
Afurtherissuetoconsideristhatwefailedtodetectparasitespeciesthatgenuinely
infectedthepopulationbecausetheydidnotsurvivewellinthesearchaeologicalcontexts.
ThehotdryclimateoftheMediterraneanregionmaynotbeasconducivetothesurvivalof
parasiteeggscomparedwithcooler,moisterarchaeologicalsoilsinnorthernEurope,for
example.Theeffectoftaphonomyismostlikelytoberelevantforspecieswiththinand
fragileeggs,suchaspinwormorhookworm(Bouchetetal.,2003).Theabsenceofprotozoa
causingdysenteryisunlikelytoreflectgenuineabsenceofdysenteryinthepopulation
throughouttheRomanperiod,asELISAtestsforGiardiaduodenaliswerepositivefrom
RomanperiodSagalassos,indicatingthatthisorganismwaspresentintheregionatthe
time(Williamsetal.,2017).Itispossiblethatprotozoawerepresentinatleastsomeofthe
populationatEphesus,butthatthesamplesweanalyseddidnotcontainahighenough
concentrationofantigentotriggerapositiveresultforGiardia.Thereisalsoevidencefor
EntamoebahistolyticaintheRomanperiodfromsitesinBelgium,France,andItaly(LeBailly
andBouchet,2015).WeshouldconsiderthattheHClneededtodisaggregatethe
mineralisedsamplesfromtheterracehouselatrinemaypotentiallyhaveinterferedwiththe
abilityoftheELISAkitstodetectprotozoaiftheywerepresent.
Afinalfactorweshoulddiscussisthepotentialuseofanti-helminthicmedicinesby
localmedicalpractitioners.GalenofPergamum(130-210CE)wasamedicalpractitioner
fromAsiaMinor,whodescribedseveraltypesofintestinalhelminthinhismedicalworks
andrecommendedarangeofmedicaltreatmentstoexpelthem(JirsaandWiniwarter,
2010).Anumberofthosemedicineshavebeenshowntohaveanti-helminthiceffectsin
laboratorystudiesandwhengiventoanimalsinfectedwithhelminths(Githiorietal.,2006;
Idrisetal.,1982;Khanetal.,2015;Mahmoudetal.,2002;Molefeetal.,2012).However,
thedegreetowhichthesemedicinesmayhavebeenusedinRomantimes,andhow
effectivetheymayhavebeeninhumans,isveryhardtoevaluateusingarchaeological
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evidenceaswehavehere.
PalaeoparasitologicalanalysisofmultiplelatrinesedimentsfromRomanEphesus
allowsustoexplorethedistributionofparasitesfromasinglesite.Interestingly,the
parasitesfoundinthepublicandprivatelatrinefromthesiteshowednooverlap.
Roundwormwasfoundaloneinthepubliclatrine,andwhipwormwasfoundaloneinthe
privatehouselatrine.Weexpectedtofindalownumberofspeciesinthehouselatrinedue
tothesmallnumberofpeopleusingit,whileweexpectedalargernumberofspeciesinthe
communalpubliclatrine,aslargenumbersofpeopleandlikelymorevisitorswouldhave
usedthiseveryday.However,onlyonespecies,roundworm,wasfoundinthesewerdrain
ofthepubliclatrine.Sincewestudiedmultiplesamplesfromthedrain,thiswouldsuggest
thatthepopulationwasgenuinelynotinfectedbyabroadrangeofparasites.Thisis
confirmedbythefactthatintheharboursediment,whereallthecitydrainsandeffluent
eventuallyendedup,onlywhipwormandroundwormeggswerefound.
Currently10speciesofintestinalparasitehavebeenidentifiedinRomanperiodsites
acrosstheempire.Ofthesespecies,whipwormandroundwormwerethemostcommon.
WhipwormandroundwormhavebeenfoundtogetheratRomanperiodsitesinAustria,
Belgium,Britain,France,Germany,Greece,Israel,andtheNetherlands(Anastasiouetal.,
2017;Aspöcketal.,1999;Bouchetetal.,2001;Boyer,1999;Gourevitchetal.,2011;Jansen
andOver,1962;Jones,1985;Jones,1987;JonesandHutchison,1991;Knightsetal.,1983;
KuijperandTurner,1992;Pike,1968;Rouffignac,1985;Roussetetal.,1996;Wilsonand
Rackham,1976;Ziasetal.,2006).However,therearealsofoursiteswhereroundwormhas
beenfoundwithoutwhipworm(Dittmaretal.,2002;Harter,2003;Aspöcketal.,2011;
Williamsetal.,2017),andsixsiteswherewhipwormhasbeenfoundwithoutroundworm
(Witenberg,1961;deMoulins,1990;Carrottetal.,1995;vanGeeletal.,2003;Heirbautet
al.,2011;Dufouretal.,2016).Thesampletypedoesnotappeartoaffectthepreservationof
thesetwoparasites,asbothspeciesoccurinlatrinesediment,mummifiedremains,pelvic
soil,andoccupationlayersediment.BasedonotherworkundertakenonRomanperiod
samples,itappearsthatitismostcommontofindbothspeciespreservedtogether,but
thereareasubstantialnumberofcaseswhereroundwormorwhipwormoccurwithoutthe
other.Thismayindicatethatincertainenvironments,oneparasitewasmoreeffectivethan
theotheratinfectingthepopulation.
13
Studyoftheharboursedimentcoreprovidestheopportunitytolookforchangeover
timeinthetypeofparasitespresent.Theeggsofwhipwormwerefoundineverycore
datingfromtheRomanperiod,withoccasionalroundworm.Wedidnotdetectanychange
inthespeciesofparasitethroughouttheRomanperiod.However,itisinterestingthatfar
morewhipwormeggswerefoundintheharboursedimentthanwasthecasefor
roundworm,withtheratiobeing19to1.Modernroundwormsproduceabout240,000eggs
perdayinthatindividual'sfaeces,andwhipwormsproducefarfewereggs,around5,000
eggsperday(WellerandNutman,2014).Asthereisnoreasontobelievethatroundworm
eggsarelesslikelytosurviveinarchaeologicalcontextsthanwhipwormeggs,thiswould
suggestthatovertimetherewerefarmorewhipwormsthanroundwormsproducingeggsin
thepopulationofEphesus.
6.Conclusions
WestudieddifferentsamplesfromacrossEphesususingmicroscopyandELISAto
determineboththespeciesofintestinalparasitethatinfectedthepopulation,aswellasany
differencesininfectionbetweenpeopleusingdifferentlatrines.Analysisofthepubliclatrine
revealedhumanroundworm,whilehumanwhipwormwasfoundintheprivatehouse
latrine.Thedominanceofwhipwormeggsoverroundworminsequentialsamplingofa
sedimentcorefromtheRomanharboursuggeststhatwhipwormwasprobablythemost
commonintestinalparasiteintheinhabitantsthroughouttheRomanperiod,andtherewas
noclearevidenceforchangeinparasitismovertime.Thefactthatboththesespeciesof
parasitearespreadbythefaecalcontaminationoffoodanddrinkhighlightsthatsanitary
issueswerethedominantfactorresultinginthespreadofparasitesinthispopulation.The
resultswerecomparedtowhatisknownaboutparasiticinfectionintherestoftheRoman
Empire,aswellastheoneotherarchaeologicalsitethathasbeenstudiedinTurkey
(Sagalassos).TheresultsfromthosesitessofarstudiedinRomanperiodAsiaMinordo
appeartocontrastwiththosefromnorthernEuropeduringthesameperiodwheremany
morespecieswerepresent.Thismaybeduetodifferencesinclimate,diet,cooking,medical
practices,andthepreservationofparasiteeggsindifferentarchaeologicalenvironments.
14
Acknowledgements
WearegratefultoTechlab©,Blacksburg,USAfordonatingtheELISAtestkits.Wewould
liketothanktheTurkishGeneralDirectorateforAntiquitiesinAnkarafortheworking
permitinEphesusandtheexportlicenceforsoilsamples.
FundingSources
ThisresearchwassupportedbyadoctoralawardfromtheSocialSciencesandHumanities
ResearchCouncilofCanada[752-2016-2085]andaTidmarshCambridgeScholarshipfrom
theCambridgeCommonwealth,EuropeanandInternationalTrustandTrinityHallCollege.
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ListofFigures
Figure1.MapshowingthelocationofEphesusinrelationtootherRomancitiesinthe
RomanprovinceofAsia(Pergamum,Sardis,andSagalassos).Image:ÖAI-ÖAW.
Figure2.MapofRomanEphesus.(A)Harbourcanal;(B)PubliclatrineatVariusbath
complex;(C)PrivatelatrineinTerraceHouse(Room34/34a).Image:ÖAI-ÖAW.
26
Figure3.DrainfromthepubliclatrineassociatedwiththeVariusbathcomplex.Imageshowsthelayersofsedimentthataccumulatedinthedrain.Image:FriederikeStock.
27
(a)
(b)
Figure4.a)PrivatelatrineinTerraceHouse2(Room34/34a).Theoriginalroomcontaining
thelatrineistheareatotheleftofthedashedline.Thepositionofthesewerdirectly
connectedtolatrineistheshadedregiontotheleftandbottomoftheimage(top).Image:
ÖAI-ÖAW.;b)mineralizedmaterialscrapedfromthedrainofthelatrine(bottom).Image:
MarissaLedger.
28
Figure5.DecorticatedroundwormeggfromthepubliclatrineatEphesus(65.6µmlongand
46.2µmwide).Scalebaris20µm.Image:MarissaLedger.
Figure6.WhipwormeggfromtheprivatehouselatrineatEphesus(50.4µmlongand27.7
µmwide).Scalebaris20µm.Image:MarissaLedger.
29
Figure7.WhipwormeggfromEphesusHarbourCanal(50.0µmlongand23.0µmwide).
Scalebaris20µm.Image:PiersMitchell.
