Note

Eioilmisdoinodasanadhoronmarix-onclosod
bacorialpopulaionAoi?00?,Cosoronoal.,
1995,Cosoronoal.,1999,Davoyoal.,?000.n
nauro,microorganismsusuallyadhoroosolidsur-
acosospociallyonholiquid-solidinoraco.Aor
aachmon, hoy orm microcolonios, produco
oxracollularpolymoricsubsancosEF8,onrapcoll
dobrisandohorspociosocolls,andormbioilm
Aoi?00?,Cosoronoal.,1995,Cosoronoal.,
1999,Gomo.oal.,?003,Toradaoal.,?003.n
gonoral,bioilm isvoryrosisanoanibioicsand
somophysicalroamonsAoi?00?,Cosoronoal.,
1995,Cosoronoal.,1999,Kumaroal.,1998,
Zoolaoal.,1994,andbioilmisconsidorodobo
asoriousproblominoodhygiono.
Thorohavoboonsomosudiosonhorolosoo-
luxpumpsindrugrosisancoandbioilmormaion
ooal.,?008,Homanoal.,?005,Mayoal.,
?009,Zhuoal.,?008.Homanoal.?005ro-
porodhaaminoglycosidoanibioicsinducobaco-
rialbioilmormaionHomanoal.,?005.ooal.
?009showodha/cc//bioilmhashighoranibi-
oicrosisanco,andhahogonooxprossionosomo
oluxpumpswasincroasodinbioilm.Mayoal.
?009showodhaanibioicsasub-inhibiorycon-
conraionsinducodhogonooxprossionosomoo-
luxpumpsin/cc//wihpEF3??,andhahoox-
prossodoluxpumpgonosconribuodohomaura-
ionobioilm.Kvisoal.?008showodhaho
addiionooluxpumpinhibiorsdocroasodho
bioilmormaionosomobacoriaincluding/cc//.
Thorosulsinhosoprovioussudiosshowodha
somomulidrugoluxpumpssoomodoconribuoo
hobioilmormaion.
nhissudywoinvosigaodhooocsodoloing
gonoscodingmulidrugoluxpumpsonhobioilm
ormaiono/cc//K-1?.Mulidrugoluxpumpgonos
in/cc//K-1?havoboonwollsudiod,andhoyaro
caogori.odino5ypos.hoATFbindingcassoo
AEC suporamily,ma|oraciliaorsuporamily
MF8,smallMDF8MF amily,rosisanco-
nodulaion-colldivisionFDamilyandmulidrug
and oxic compound oxrusionMATE amily
Lubolskioal.,?007,ishinooal.,?009.Fumps
oxcopohoAECsuporamilyaroproonmoivo
orcoFMForsodiumionmoivoorcoMATEam-
ilyypopumps.nhissudy,wousodmuanswih
doloodgonosoxcopromhoMATEamilyo/cc//
K-1?EW?5113.
Thobacoriausodworo/sc|/c|/acc//K-1?
EW?5113aa/-aa/567,/ac4ZZ
..
^/4
/ac/o4000/ac/
(
andgono-rolaodmuansdorivod
rom EW?5113woroobainodrom hoaional
nsiuooGonoics8hi.uoka,Japansupporodby
aionalEioFosourcoFro|ocG,Japan./cc//.
HoIesofMuItIdrugEffIuxPumpsonthe
BIofIImFormatIonofEscher/ch/aco//K-12
KAYOMATSUMUHA,SOlCHlFUHUKAWA

,
HlHOKAZUOGlHAHA,ANDYASUSHlMOHlNAGA
/oa/^/c//ccJ//csc/^ca^J//c/c|^c/c,c//c///cscac5c/^cs
^/|c^|^/s//,100/a/^c/a//sana?b?00'aoa^
Focoivod1?ovombor,?010/Accopod11March,?011
WegenetIcaIIyanaIyzedtheroIesofmuItIdrugeffIuxpumpsonthebIofIIm formatIonof
Escher/ch/aco//K-12BW25113.Weused22mutantsmIssIngvarIousgenesreIatedtothe
muItIdrugeffIuxpumpsandfoundthatthebIofIImformatIonofemr0,emrE,emrK,acr0,acre
andmdlE-deIetedmutantswasextremeIydecreased.TheseresuItsIndIcatethatsome
muItIdrugeffIuxpumpssIgnIfIcantIycontrIbutetothebIofIImformatIonofEco//.
/,ncJsEioilm//sc|/c|/acc///Mulidrugoluxpump.
ioconlrolScience,!011,Vol.16,No.!,67!

csoc^J/^aa/|c1//a-14004Z/a//
/aa/anaa/s^/|c^aac/o
/cc//collsworogrowninLEbrohMFEiomodicals,
OH,U8A)ovornigh.
Esimaionobioilmormaionwasporormodac-
cordingohomohoddoscribodproviouslyJackson
oal.,?00?,arisawaoal.,?005)wihslighmodii-
caions.Ovornighculurosworoinoculaodinorosh
LEmodiumorDavismodiumK?HFO47g,KH?FO4?
g,Mg8O47H?O0.1,H4)?8O41g,8odiumcirao
0.5g,Glucoso?g)a1.100raiooal?00;l)and
grownin96-wollmicroiorplaosa30or16o48
h.nhissudy,woadopodpolysyronomicroior
plaoswihouForvair8cioncos,U.K.)orwihTFF
TochnoFlasicFroducsAG,8wi.orland)corona
dischargoroamon.Thohydrophobicpolysyrono
plaoswihoucoronadischargoroamonarousually
usodinbacorialbioilmormaionassaysandhohy-
drophilicpolysyronoplaoswihcoronadischargo
roamonarousuallyusodinculuringhumanand
animalcolls.
Eioilmormaionwasosimaodbymoasuringho
crysalviolosainingohocollsaachodoho
microiorplaowolls.Aordiscardinghomodiumin
homicroiorplaowolls,howollswororinsodwico
wihwaor.Thon,0.1%crysalvioloMorck,Tokyo)
wasaddodohowollsorsaininghoaachodcolls.
Aordiscardinghocrysalviolosoluionandrinsing
howollswihwaor,crysalviolowasoxracodby
70% ohanolsoluion,andhasoluionwasrans-
orrodoanowmicroiorplao,andhoabsorbanco
a590nmwasmoasurodusingaplaoroadorE-
max,ihonMolocularDovicos,Tokyo).Foroachox-
porimon,corrocionorbackgroundsainingwas
madobysubracinghovaluoorcrysalviolobound
oun-inoculaodconrols.Aloasoighmicroior
plaowollsworousodoroachsamplo,andoachox-
porimonwasporormodaloaswico.Thosandard
doviaionsocrysalvioloabsorbancoorhooigh
microiorplaowollswororom1/?0o1/5ohoir
avoragovaluos,andhosandarddoviaionsorhroo
ormoroindopondonoxporimonsworoalsorom
1/?0o1/5ohoiravorago.
Alloxporimonsworodonoinduplicao.Thodaa
prosonodarohomoansohrooormorooxpori-
mons.8igniicandioroncosworodoorminodby
8udon's/osF<0.05).
Wousod/cc//K-1?AM1?46F

),MG1655
F

o|1)andW3110F

/^^/^/)1o|
1)sainsinprovioussudiosKawaraioal.,?009,
arisawaoal.,?005),andhoabovosrainsormod
bioilmvorywollbowoonapprox.0.?o0.9A590)or
?4hinLEmodiumbyusinghohydrophobicpolysy-
rono96-wollplao.Howovor,/cc//K-1?EW?5113
/aa/aa/)567,//ac4ZZ
..
^/4)/ac/o
4000/ac/
(
))ormodlossbioilm byapprox.0.05
A590)inhosamocondiions.Thoroorowoinvosi-
gaodhocondiionsorhoimprovodbioilmorma-
iono/cc//K-1?EW?5113.
Airs,hobioilm ormaiono/cc//K-1?
EW?5113wascomparodonhydrophilicplaoswih
coronadischargoroamon)andhydrophobicplaos
wihoucoronadischargoroamon)inwoyposo
culuromodiumLEandDavismodium)avarious
culuroimosFig.1).Thobioilmormaiononhohy-
drophilicplaoswasrolaivolyhighorhanhaonho
hydrophobicpolysyronoplaos,whichhadbooncon-
vonionallyusodinbacorialbioilmormaionassays.
Espociallyhobioilm ormaiono/cc//K-1?
EW?5113inDavismodiumonhohydrophilicplao
washighorirrospocivooculivaionimo.Howovor,
hocombinaionoDavismodium andhydrophilic
plaoswasnosuiabloorhobioilmassaybocauso
hoamounsobioilmormodworoobviouslyunsa-
blo |udging rom ho largo sandard doviaions
Fig.1E).Onhoohorhand,/cc//K-1?EW?5113
ormodbioilminsabloandadoquaoamounsinLE
modiumonhydrophobicplaosaor40hculivaion
Fig.1A).Horoaor,hobioilmormaiono/cc//K-
1?EW?5113wasassayodbyculivaioninLEmo-
diumonhydrophobicplaosor40h.Thosorosuls
indicaohahydrophobicorhydrophilicproporioso
hosolidsuracosigniicanlyaochobioilmorma-
iono/cc//.
Thooocodoloingogonosrolaodomulidrug
oluxpumpsonhobioilmormaiono/cc//K-1?
EW?5113wasinvosigaodundorhoabovodoor-
minodassaycondiions.Wousod??muansha
woromissingvariousgonosrolaodomulidrugo-
S.FUkUKAWAFTAL.
.Eocsohoplaowihorwihoucoronadis-
chargoroamon),modiaandculivaionimoonho
bioilmormaiono/cc//K-1?EW?5113.
LE.LEmodium,Davis.Davismodium.
luxpumps.Allgono-doloodmuansusodinhis
sudyshowodsigniicanlydocroasodbioilmorma-
ioncomparodwihhoparonsrain/cc//K-1?
EW?5113.Espocially,muanswihou//
/ac/ac/andJ//showodoxromolylowor
bioilmormaionFig.?.Thososixgonosbolongo
hogonoscodinghoproonmoivoorcoFMF
pumps.Fromhosorosuls,FMFpumpsworoconsid-
orodohavoimporanrolosinhobioilmormaion
o/cc//.
Frovious sudios suggosod hasomo olux
pumpsrolaodohooluxoanibioicsconribuos
ohobioilmormaiono/cc//Lucasoal.,?005,
Mayoal.,?009.Ourrosulsparlycorrospondodo
horosulsohosoprovioussudios.Frocisomocha-
nismsohoconribuionohosooluxpumpsonho
/cc//bioilmormaionshouldboclariiodinhou-
uro.Thoymighconribuoohobioilmormaion
/ahooxpororimporosomosubsancoshaaro
nocossaryorharmulorbioilmormaion.
ACKNOWLEDGMENT
Wowouldlikoohankhoaionalnsiuoo
Gonoics8hi.uoka and aionalEioFosourco
Fro|ocG,Japan./cc//orgivingusho/cc//
srainsusodinhissudy.
HEFEHENCES
Aoi,Y.?00?/^s//aidoniicaionomicroorganismsin
bioilmcommuniios.'//csc///c^.,94,55?-556.
Cosoron,J.W.,Lowandowski,Z.,Caldwoll,D.E.Korbor,D.
F.,andLappin-8co,H.M.1995Microbialbioilms.
^^^///cc//c/.,49,711-745
Cosoron,J.W.,8owar,F.8.,andGroonborg,E.F.1999
Eacorialbioilms.acommoncausooporsisoninoc-
ions.5c/^c,284,1319-13??
Davoy,M.E.,andO'Toolo,G.A.?000Microbialbioilms.
romocologyomoloculargonoics.//cc//c//c///c/
/.,64,847-867
Gigho,J.?001auralcon|ugaivoplasmidsinducobac-
orialbioilmdovolopmon.^a/a,412,44?-445.
Gomo.,M.A.,Galvo.,J.M.,Honoria,E.,andGon.alo.-
Lopo.,J.?003nluoncooohanolconconraionon
bioilm bacorialcomposiionrom adoniriyingsub-
morgodilorusodorconaminaodgroundwaor.'
//csc///c^.,95,?45-?51.
Homan,L.F.,D'Argonio,D.A.,MacCoss,M.J.,Zhang,Z.,
Jonos,F.A.,andMillor,8..?005Aminoglycosidoani-
bioicsinducobacorialbioilm ormaion.^a/a,436,
1171-1175.
o,A.,Taniuchi,A.,May,T.,Kawaa,K.,andOkabo,8.
?009ncroasodanibioicrosisancoo/sc|/c|/acc//
inmaurobioilms.^oo//^//cc//c/.,75,4093-100.
Jackson,D.E.,8u.uki,K.,Oakord,L.,8imocka,J.W.,Har,
M.E.,andFomoo,T.?00?Eioilmormaionanddis-
porsalundorhoinluoncoohoglobalrogulaorCsrAo
/sc|/c|/acc//'/ac//c/.,184,?90-301.
Kawarai,T.,Furukawa,8.,arisawa,.,Hagiwara,C.,
Ogihara,H.,andYamasaki,M.?009Eioilmormaion
byEschorichiacoliinhyporonicsucrosomodia.'
//csc///c^.,107,630-635.
MULTDkUCFFFLUXUMANDFLMFkMATN 71
FlG.2.EocsoMDF-rolaodgonodoloiononhobioilmormaiono/cc//
K-1?EW?5113.
AEC.ATFbindingcassoosuporamily,MF8.Ma|oraciliaorsuporamily,
8MF.8mallMDFamily,FD.Fosisanco-nodulaion-colldivisionamily.
Kumar,C.G.,andAnand,8.K.19988igniicancoomicro-
bialbioilmsinoodindusry.aroviow./^/'/ccJ
//cc//c/.,,?9-?7.
Kvis,M.,Hancock,V.,andKlomm,F.?008nacivaion
ooluxpumpsabolishosbacorialbioilm ormaion.
^oo//^/c^//cc//c/.,,7376-738?.
Lubolski,J.,Konings,W..,andDriosson,A.J.?007
DisribuionandphysiologyoAEC-yporansporors
conribuingomulidrugrosisancoinbacoria.//cc//c/
/c///c//.,,463-476.
May,T.,o,A.,andOkabo,8.?009nduciono
mulidrug rosisanco mochanism in /sc|/c|/a cc//
bioilmsbyinorplaybowoonoracyclinoandampicillin
rosisanco gonos. ^^///cc//a/ ^^/s a^J
|c/|ao,,,46?8-4639.
arisawa,.,Furukawa,8.,Ogihara,H.,andYamasaki,M.
?005Esimaionohobioilmormaiono/sc|/c|/a
cc//K-1?byhocollnumbor.'//csc///c^.,,78-
80.
ishino,K,ikaido,E.,and Yamaguchi,A.?009
Fogulaionandphysiologicaluncionomulidrugolux
pumpsin/sc|/c|/acc//and5a/c^//a//cc|/
//co|,s^c/a,,834-843.
Torada,A.,Hibiya,K.,agai,J.,Tsunoda,8.,andHiraa,A.
?003irogonromovalcharacorisicsandbioilm
analysisoamombrano-aoraodbioilmroacorapplica-
bloohigh-sronghnirogonouswasowaorroamon.'
//csc///c^.,,170-178.
Zhu,X.,Long,F.,Chon,Y.,Knochol,8.,8ho,O.,and8hi,X.
?008ApuaivoAECranspororisinvolvodinnogaivo
rogulaion o bioilm ormaion by //s//a
c^cc,/c^s^oo//^//cc//c/.,,7675-7683.
Zoola,E.A.,and8asahara,K.C.1994Microbialbioilms
inhooodprocossingindusry--shouldhoyboacon-
corn?/^/'/ccJ//cc//c/.,,1?5-148.
S.FUkUKAWAFTAL.