biofilms-091008210814-phpapp02 imp.ppt
TRANSCRIPT
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Beginning of MicrobesBeginning of Microbes
Bacteria frst appearedBacteria frst appearedon earth about 3.6on earth about 3.6billion years ago, longbillion years ago, longbeore the appearancebeore the appearance
oo Homo sapiensHomo sapiensaroundaround100,000 years ago..100,000 years ago..Van LeeuwenhoekVan Leeuwenhoekwas the frst person towas the frst person tovisualize, graphicallyvisualize, graphicallyillustrate, and labelillustrate, and label"anialcules" !bacteria"anialcules" !bacteriathat he ound inthat he ound inplaqueplaquescraped ro his ownscraped ro his ownteeth.teeth.
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#he ter biofl was introduced to designate the#he ter biofl was introduced to designate the
thinlayered condensations o icrobesthinlayered condensations o icrobes
!e.g.bacteria, ungi, protozoa that ay occur on!e.g.bacteria, ungi, protozoa that ay occur on
various surace structures in nature. $ree%&oatingvarious surace structures in nature. $ree%&oating
bacteria e'isting in an a(ueous environent,so%bacteria e'isting in an a(ueous environent,so%
calledplan)tonicicroorganiss,are acalledplan)tonicicroorganiss,are a
prere(uisite or biofl oration. *uch flsprere(uisite or biofl oration. *uch fls
ay thus becoe established on any organic oray thus becoe established on any organic or
inorganic surace substrate where plan)tonicinorganic surace substrate where plan)tonicicroorganiss prevail in a water%basedicroorganiss prevail in a water%based
solution.solution.
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Despite the discovery of microbial biolms as far back asthe 17thcentury, scientists have largely focused their attentions onthe solitary,or planktonic, forms of microorganisms.
In nature, hoever, most microorganisms live together inlargecommunities attached to a surface+ere, bacteria ree in saliva !plan)tonic organiss serve as thepriary source or the organization o this specifc biofl !1
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Biofls were already discovered by one o theBiofls were already discovered by one o the
frst icrobiologist, the utch -ntonie vanfrst icrobiologist, the utch -ntonie van
eeuwenhoe) in 16/0s, but the actualeeuwenhoe) in 16/0s, but the actual
brea)through o the phenoenon had to waitbrea)through o the phenoenon had to wait
another 3 years until 2osterton andanother 3 years until 2osterton and
colleagues published their wor) on +owcolleagues published their wor) on +ow
Bacteria *tic)4 in 15. *ince 15 theBacteria *tic)4 in 15. *ince 15 the
research within biofl bacteria has e'ploded.research within biofl bacteria has e'ploded.
#he inherent high tolerance to antiicrobials#he inherent high tolerance to antiicrobials
and iune cells e'plain its vast iplication inand iune cells e'plain its vast iplication in
chronic inections.chronic inections.
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Bacteria are ound in at least two distinct states 7Bacteria are ound in at least two distinct states 7
either as plan)tonic or sessile cells. 8lan)toniceither as plan)tonic or sessile cells. 8lan)tonic
cells are classically defned as ree &owingcells are classically defned as ree &owing
bacteria in suspension4 as opposed to the sessilebacteria in suspension4 as opposed to the sessile
state !the so called biofl9 a structuredstate !the so called biofl9 a structuredcounity o bacterial cells enclosed in a sel%counity o bacterial cells enclosed in a sel%
produced polyeric atri' and adherent to anproduced polyeric atri' and adherent to an
inert or living surace4 as stated by 2osterton andinert or living surace4 as stated by 2osterton and
cowor)ers !1555. :n the above section sessilecowor)ers !1555. :n the above section sessilegrowing bacteria were defned as an asseblagegrowing bacteria were defned as an asseblage
o cells ebedded 4in a sel%produced polyerico cells ebedded 4in a sel%produced polyeric
atri'atri'
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efnition o a Bioflefnition o a Biofl
Biofl can be defned as a sessileBiofl can be defned as a sessile
ulticellular icrobial counityulticellular icrobial counity
characterised by cells that are frlycharacterised by cells that are frly
attached to a surace and eneshedattached to a surace and eneshed
in a sel produced atri' oin a sel produced atri' o
e'tracellular polyeric substancee'tracellular polyeric substance
usually polyssacharideusually polyssacharide
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efnition o a Bioflefnition o a Biofl
BioflsBioflsare counities oare counities o
icroorganiss in a atri' that ;oins theicroorganiss in a atri' that ;oins the
together and to living or inert substratestogether and to living or inert substrates
BioflsBioflsare surace%attached counitiesare surace%attached counitieso bacteria, encased in an e'tracellularo bacteria, encased in an e'tracellular
atri' o secreted proteins,atri' o secreted proteins,
carbohydrates, and
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Biofl are not erely passiveBiofl are not erely passive
asseblage o bacterial cells that areasseblage o bacterial cells that are
stuc) to surace they are structurallystuc) to surace they are structurally
and dynaically organized cople'and dynaically organized cople'
biologic syste.biologic syste.
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Biofl orationBiofl oration
#he frst step is the#he frst step is the attachentattachento theo thebacterial cells to the selected abiotic orbacterial cells to the selected abiotic or
biotic suracebiotic surace
:nitial attachent is ediated through:nitial attachent is ediated throughwea) reversible van der >aalswea) reversible van der >aalsinteractionsinteractions
between the cell surace and thebetween the cell surace and thesubstratu, which can lead to strongersubstratu, which can lead to strongeradhesion receptor ediated attachentadhesion receptor ediated attachent
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Bacterial cell surace structures such asBacterial cell surace structures such as&agella&agella,, fbriaefbriae,, 8*8*, and, ande'opolysaccharidese'opolysaccharidesparticipate inparticipate inirreversibleirreversibleinteractionsinteractions
#he second step corresponds to the#he second step corresponds to the
developent odevelopent o icro%coloniesicro%coloniesprooted by the growth and division oprooted by the growth and division othe frst attached cells !priarythe frst attached cells !priarycolonizerscolonizers
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#he icro%colonies progressively#he icro%colonies progressively
enlargeenlarge
and coalesce to or theand coalesce to or the
frstfrst
layer o cellslayer o cells covering the suracecovering the surace
>hen>hen ultiple layersultiple layers o cells pile upo cells pile upon the surace, the third step o theon the surace, the third step o the
oration is obtainedoration is obtained
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indicated by the presence oindicated by the presence o a aturea aturebioflbiofl characterized by the presence ocharacterized by the presence oacro%coloniesacro%coloniessurrounded bysurrounded by waterwater
channelschannels that help distribute nutrientsthat help distribute nutrientsand signaling oleculesand signaling olecules
$inally, to survive when$inally, to survive when nutrientsnutrients
becoe liitedbecoe liited, or siply to spread and, or siply to spread andcolonize to other niches, soe bioflcolonize to other niches, soe bioflcells cancells can detachdetachindividually or in clupsindividually or in clups
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copositioncoposition
1/? bacterial icrocolonie1/? bacterial icrocolonie
/? e'tracellular polysaccharide/? e'tracellular polysaccharide
atri'atri'
Both o these are separated by waterBoth o these are separated by water
channelschannels
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Biofl *tructureBiofl *tructure
biofls display a coon attribute, thebiofls display a coon attribute, thebioflbioflatri'atri'..
2ontrary to ree%&oating plan)tonic cells,2ontrary to ree%&oating plan)tonic cells,biofl cells are ebedded in a sel producedbiofl cells are ebedded in a sel producede'tracellular atri', thee'tracellular atri', the e'tracellulare'tracellularpolyeric substancepolyeric substance!@8*, that!@8*, that holds theholds thetogethertogether
Biofls are coposed o aboutBiofls are coposed o about 07/? @8*07/? @8*!by volue and only!by volue and only 1/70? cells1/70? cells !by volue!by volue
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-lthough the-lthough the @8*@8*ay vary in cheicalay vary in cheical
and physical properties, its a;orand physical properties, its a;or
coponents arecoponents are polysaccharidespolysaccharides,,
proteinsproteins, and, and e'tracellular =-e'tracellular =-
#he @8* plays a a;or role in#he @8* plays a a;or role in
aintaining the integrityaintaining the integrityo the bioflo the biofland can coner other benefcialand can coner other benefcial
properties.properties.
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*ince the @8* is also*ince the @8* is also highly hydratedhighly hydrated, it, it
can preventcan prevent desiccationdesiccationin soe naturalin soe natural
bioflsbiofls
#he @8* can also act as a#he @8* can also act as a diAusiondiAusion
barrierbarrier, preventing, preventing to'ic substancesto'ic substancessuch as antibiotics and disinectantssuch as antibiotics and disinectants
ro reaching their targetro reaching their target
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2ell heterogeneity within2ell heterogeneity within
bioflsbiofls
ost biofls ound in nature areost biofls ound in nature are
polyicrobialspolyicrobials, where diverse, where diverse
species e'pressing ultiplespecies e'pressing ultiple
phenotypes are involvedphenotypes are involved
#he ost aazing act is that even#he ost aazing act is that even
in ain a ono%speciesono%speciesbiofl,biofl,
phenotypic heterogeneityphenotypic heterogeneitye'istse'ists
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2ell diAerentiation2ell diAerentiation in biofls ayin biofls ay
depend on thedepend on the local environentallocal environental
conditions surrounding the cellsconditions surrounding the cells
iAerent concentrationiAerent concentrationgradients ogradients o
o'ygeno'ygen,, nutrientsnutrients,, ionsions, and, and cheicalscheicals
create a wide variety o icrohabitatscreate a wide variety o icrohabitatsproviding conditions suitable orproviding conditions suitable or
bacterial colonizationbacterial colonization
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2ells located in the2ells located in the upper biofl layersupper biofl layersconsue all available o'ygen andconsue all available o'ygen and growgrowaerobicallyaerobically, while an, while an anaerobicanaerobicicro%nicheicro%nichedevelopeddeveloped underneathunderneaththe aerobic layerthe aerobic layer
C'ygen% and nutrient depletedC'ygen% and nutrient depleted regions areregions areound at theound at the botto layersbotto layers o the bioflo the biofl
structure and under these circustances,structure and under these circustances,ost o the sessile cells are etabolicallyost o the sessile cells are etabolicallyinactiveinactiveoror deaddead
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2ounication language in2ounication language in
bioflbiofl
- ew years ago, >atnic) D Eolter- ew years ago, >atnic) D Eolter
proposed the interesting idea thatproposed the interesting idea that
biofls can be copared to citiesbiofls can be copared to cities
:n these cities o icrobes,:n these cities o icrobes,
icroorganiss are consideredicroorganiss are considered
socialsocial4 organiss4 organiss able toable to
counicatecounicatewith one anotherwith one another
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Fsing diAerent cheical languages,Fsing diAerent cheical languages,
bacteria learn about their currentbacteria learn about their current cellcell
populationpopulation and deterine when theyand deterine when they
havehave reached a critical assreached a critical ass
Fsing that inoration, bacteria canFsing that inoration, bacteria can
thusthus odiy their behaviorodiy their behavior to carry outto carry outprocesses that would re(uireprocesses that would re(uire any cellsany cells
acting in con;unction to be eAectiveacting in con;unction to be eAective
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2ell%to%cell counication2ell%to%cell counication is generallyis generallycarried out bycarried out by diAusible signaldiAusible signaloleculesolecules produced and released byproduced and released by
bacteriabacteria
>hen bacteria are growing within a>hen bacteria are growing within abioflbiofl, they, they secrete signalingsecrete signalingoleculesolecules !auto%inducers that increase!auto%inducers that increasein concentration as a unction oin concentration as a unction obacterialbacterial cell densitycell density
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:n a process called:n a process called (uoru sensing(uoru sensing,,
bacteria counicate with onebacteria counicate with one
another by usinganother by using auto%inducersauto%inducerstoto
regulate their gene e'pression inregulate their gene e'pression inresponseresponseto &uctuations in theto &uctuations in the cellcell
population densitypopulation density
#wo types#wo types o (uoru%sensing systeso (uoru%sensing systes
are recognized in bacteria9are recognized in bacteria9
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intra%speciesintra%speciescounication andcounication and
inter%speciesinter%speciescounicationcounication
uringuringintra%speciesintra%species counication,counication,
several auto%inducersseveral auto%inducershave beenhave beenidentifedidentifed
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Gra%negativeGra%negativebacteria usually usebacteria usually use
acyl hooserine lactone !-+acyl hooserine lactone !-+ asas
signal olecules, whilesignal olecules, while Gra%positiveGra%positivebacteria utilizebacteria utilize sall peptidessall peptides
uringuring inter%speciesinter%speciescounication,counication,bacteria usebacteria use autoinducer% !-:%autoinducer% !-:%
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2ells located in the2ells located in the upper biofl layersupper biofl layersconsue all available o'ygen andconsue all available o'ygen and growgrowaerobicallyaerobically, while an, while an anaerobicanaerobicicro%nicheicro%nichedevelopeddeveloped underneathunderneaththe aerobic layerthe aerobic layer
C'ygen% and nutrient depletedC'ygen% and nutrient depleted regions areregions areound at theound at the botto layersbotto layers o the bioflo the biofl
structure and under these circustances,structure and under these circustances,ost o the sessile cells are etabolicallyost o the sessile cells are etabolicallyinactiveinactiveoror deaddead
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:n soe cases, (uoru sensing does:n soe cases, (uoru sensing doesnot see to be involved in bioflnot see to be involved in bioflstructural developentstructural developent
while or other species, there iswhile or other species, there isevidence that (uoru sensing isevidence that (uoru sensing isiportant or theiportant or the attachentattachentoo
bacteria to the surace, thebacteria to the surace, the aturationaturationo the biofl, or the control o eventso the biofl, or the control o eventsleading to theleading to the dispersiondispersiono cellso cells
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ulti%drug tolerance oulti%drug tolerance o
bioflsbiofls - signifcant characteristic o icrobial- signifcant characteristic o icrobial
biofls is theirbiofls is their high%level drughigh%level drug
tolerancetolerance
Bacterial biofls have been shown toBacterial biofls have been shown to
have ahave a 100% to 1,000% old100% to 1,000% old increasedincreased
tolerance toward antibiotics intolerance toward antibiotics incoparison to their ree%swiingcoparison to their ree%swiing
counterpartscounterparts
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iAusion barrier iparted by the @8* atri'iAusion barrier iparted by the @8* atri'
8henotypic heterogeneity8henotypic heterogeneity
Gene #ranserringGene #ranserring
*ecretion o antibiotic degrading enzyes*ecretion o antibiotic degrading enzyes
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8rotective role o @8* atri'8rotective role o @8* atri'
- priary unction that has been- priary unction that has been
attributed to the biofl atri' isattributed to the biofl atri' is
protectionprotection,,
*everal studies have shown that the*everal studies have shown that the
@8* atri' can act as an ipereable@8* atri' can act as an ipereable
barrier tobarrier to liit antiicrobialliit antiicrobialpenetrationpenetration, thereby protecting the, thereby protecting the
biofl cellsbiofl cells
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*uch protection can be due either to*uch protection can be due either to
physical hindrancephysical hindrancein antiicrobialin antiicrobial
diAusion or todiAusion or to direct bindingdirect bindingo theo the
antibiotics by the @8* atri'antibiotics by the @8* atri'
Fpon antibiotic treatent, cells at theFpon antibiotic treatent, cells at the toptopo the li(uid7 biofl interaceo the li(uid7 biofl interace diediedue todue to
their closer e'posure, while bacteriatheir closer e'posure, while bacteriaebeddedebedded deepdeepinside the biofl are ableinside the biofl are able
toto survivesurvive
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-nionic @8*-nionic @8* atri' can bind andatri' can bind and
se(uester to'icse(uester to'ic cationic heavycationic heavy
etalsetals,, cationic antiicrobialcationic antiicrobial
peptidespeptides, and, and positively%chargedpositively%charged
antibioticsantibiotics!e.g. ainoglycosides!e.g. ainoglycosides
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8henotypic heterogeneity8henotypic heterogeneity
8henotypic8henotypic heterogeneityheterogeneityoccurs withinoccurs withinbiofls notably due tobiofls notably due to diAerentdiAerentconcentration gradientsconcentration gradients
2ells localized at the2ells localized at the botto layersbotto layers o theo thebiofl are norally ound in a growthbiofl are norally ound in a growthstage analogous tostage analogous to stationary%phasestationary%phase
plan)tonic cells, while the physiology oplan)tonic cells, while the physiology ocells localized at thecells localized at the toptopsurace layers issurace layers issiilar tosiilar to e'ponentially growinge'ponentially growing plan)tonicplan)toniccellscells
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icrobial cells, especially those in theicrobial cells, especially those in thedeeper layersdeeper layers o the biofls whereo the biofls wherenutrients and o'ygen are liited, arenutrients and o'ygen are liited, are
associated with aassociated with a lower growth ratelower growth rate
#his#his reduced etabolic activityreduced etabolic activity ightightaccount or the enhancedaccount or the enhanced tolerancetolerance
toward antibioticstoward antibioticsthat target bacterialthat target bacterialcellular processes such as =-cellular processes such as =-replication or translationreplication or translation
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2onventional2onventional antibioticsantibioticsused to treatused to treat
inections are ostly eAective atinections are ostly eAective at )illing)illingrapidly growingrapidly growing cellscells
#he#he decreased etabolic activitydecreased etabolic activity oo
cells ound within the deeper bioflcells ound within the deeper biofl
layers ay thus contribute tolayers ay thus contribute to antibioticantibiotictolerancetolerance and the persistence oand the persistence o
biofl inectionsbiofl inections
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Gene #ranserGene #ranser
Biofls also provide anBiofls also provide an ideal nicheideal niche
or theor the e'changee'changeoo
e'trachroosoal =- !e'trachroosoal =- !plasidsplasids
2on;ugation !the echanis o2on;ugation !the echanis o
plasid transer occurs atplasid transer occurs at a greatera greater
raterate between cells in biofls thanbetween cells in biofls than
between plan)tonic cellsbetween plan)tonic cells
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*ecretion o antibiotic*ecretion o antibiotic
degrading enzyesdegrading enzyes
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Bio lm constitutesBio lm constitutes
--biolmbiolmis an aggregate ois an aggregate o
icroorganiss in which cells are stuc)icroorganiss in which cells are stuc)
to each other and
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Biolm is a comple!Biolm is a comple!
substance.substance. - biofl is a cople'- biofl is a cople'
aggregation oaggregation o
icroorganiss growingicroorganiss growing
on a solid substrate.on a solid substrate.
Biofls areBiofls arecharacterized bycharacterized by
structural heterogeneity,structural heterogeneity,
genetic diversity,genetic diversity,
cople' counitycople' counity
interactions, and aninteractions, and ane'tracellular atri' oe'tracellular atri' o
polyeric substances.polyeric substances.
Biofls ound in =atureBiofls ound in =ature
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Biofls ound in =atureBiofls ound in =ature
everywhere where is there iseverywhere where is there is
oistureoisture
More properly known asMore properly known as bioflmbioflm,,
slime cities thrive wherever there isslime cities thrive wherever there is
water - in the kitchen, on contactwater - in the kitchen, on contact
lenses, in the gut linings of animals.lenses, in the gut linings of animals.When the urban sprawl is extensive,When the urban sprawl is extensive,
bio lms can be seen with the nakedbio lms can be seen with the naked
eye, coating the inside of water pipeseye, coating the inside of water pipesor dangling slippery and green fromor dangling slippery and green from
plumbingplumbing."." "#oghlan 1$$%&"#oghlan 1$$%&
i l hBi l h
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Biolm supports theBiolm supports the
Bacterial grothBacterial groth Biofl are a coon ode o bacterialBiofl are a coon ode o bacterial
growth in nature and their presence has angrowth in nature and their presence has anenorous ipact on any aspects o ourenorous ipact on any aspects o ourlives, such as sewage treatent, corrosion olives, such as sewage treatent, corrosion o
aterials, ood containation duringaterials, ood containation duringprocessing, pipe collapse, plant%processing, pipe collapse, plant%icroorganiss interaction in the biosphere,icroorganiss interaction in the biosphere,the oration o dental pla(ue, thethe oration o dental pla(ue, the
developent o chronic inections in livedevelopent o chronic inections in livetissue !astitis, Ctitis, pneuonia, urinarytissue !astitis, Ctitis, pneuonia, urinaryinections, osteoyelitis or probles relatedinections, osteoyelitis or probles relatedto edical iplants.to edical iplants.
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'ormation of Biolms'ormation of Biolms
Biofls ay orBiofls ay or
on living or non%on living or non%
living suraces,living suraces,
and represent aand represent aprevalent ode oprevalent ode o
icrobial lie inicrobial lie in
natural, industrialnatural, industrial
and hospitaland hospital
settingssettings
Bi fl i - ibi iBi fl i - tibi ti
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Biofls increases -ntibioticBiofls increases -ntibiotic
resitanceresitance
>ith>ith
icroorganissicroorganiss
are highlyare highlyresistant toresistant to
antiicrobialantiicrobial
treatent and aretreatent and are
tenaciously boundtenaciously boundto the suraceto the surace
h i i Bi fl h i i Bi fl
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echanisis o Bioflechanisis o Biofl
orationoration 'ormation of a biolm'ormation of a biolm
begins ith thebegins ith theattachment of free(attachment of free()oating microorganisms)oating microorganismsto a surface. *hese rstto a surface. *hese rstcolonists adhere to thecolonists adhere to thesurface initially throughsurface initially through
eak, reversible van dereak, reversible van der+aals forces.+aals forces.If theIf thecolonists are notcolonists are notimmediatelyimmediatelyseparated from theseparated from thesurface, they cansurface, they can
anchor themselvesanchor themselvesmore permanentlymore permanentlyusing cell adhesionusing cell adhesionstructures such as pilistructures such as pili
$actors :n&uencing Hate and$actors :n&uencing Hate and
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$actors :n&uencing Hate and$actors :n&uencing Hate and@'tent o Biofl $oration@'tent o Biofl $oration
:ndwelling edical device when containated:ndwelling edical device when containated
with icroorganiss, several variableswith icroorganiss, several variables
deterine whether a biofl develops. $irstdeterine whether a biofl develops. $irst
the icroorganiss ust adhere to thethe icroorganiss ust adhere to the
e'posed suraces o the device long enough toe'posed suraces o the device long enough tobecoe irreversibly attached. #he rate o cellbecoe irreversibly attached. #he rate o cell
attachent depends on the nuber and typesattachent depends on the nuber and types
o cells in the li(uid to which the device iso cells in the li(uid to which the device is
e'posed, the &ow rate o li(uid through thee'posed, the &ow rate o li(uid through thedevice, and the physicocheicaldevice, and the physicocheical
characteristics o the suracecharacteristics o the surace
* h l d t d* h l d t d
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*echnology understands*echnology understands
Biolms betterBiolms better #echnological progress in#echnological progress in
icroscopy, olecularicroscopy, olecular
genetics and genoegenetics and genoe
analysis has signifcantlyanalysis has signifcantly
advanced ouradvanced ourunderstanding o theunderstanding o the
structural and olecularstructural and olecular
aspects o biofls,aspects o biofls,
especially o e'tensivelyespecially o e'tensively
studied odelstudied odelorganiss such asorganiss such as
PseudomonasPseudomonas
aeruginosaaeruginosa..
-t i Bi l-t i Bi l
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-teps in Biolm-teps in Biolm
DevelopmentDevelopment Biofl developent canBiofl developent can
be divided into severalbe divided into several
)ey steps including)ey steps including
attachent, icroattachent, icrocolony oration, bioflcolony oration, biofl
aturation andaturation and
dispersionI and in eachdispersionI and in each
step bacteria ay recruitstep bacteria ay recruit
diAerent coponentsdiAerent coponents
and olecules includingand olecules including
&agella, type :J pili, =-&agella, type :J pili, =-
and e'o polysaccharides.and e'o polysaccharides.
-t f bi l-t f bi l
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-tages of biolm-tages of biolm
developmentdevelopment..
-teps in Biolm-teps in Biolm
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-teps in Biolm-teps in Biolm
formationformation
B t i i t d ithBacteria associated ith
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Bacteria associated ithBacteria associated ith
Biolms dierBiolms dier Bacteria living in a biofl can haveBacteria living in a biofl can have
signifcantly diAerent properties rosignifcantly diAerent properties roree%&oating bacteria, as the dense andree%&oating bacteria, as the dense andprotected environent o the fl allowsprotected environent o the fl allowsthe to cooperate and interact in variousthe to cooperate and interact in variousways. Cne beneft o this environent isways. Cne beneft o this environent isincreased resistance to detergents andincreased resistance to detergents andantibiotics, as the dense e'tracellularantibiotics, as the dense e'tracellularatri' and the outer layer o cells protectatri' and the outer layer o cells protectthe interior o the counity.the interior o the counity.
Bi fl ; Biofls a;or cause o
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Biofls a;or cause oBiofls a;or cause o
=osocoial inections=osocoial inections icrobial biofls,icrobial biofls,
which oten arewhich oten are
ored byored by
antiicrobial%antiicrobial%
resistantresistant
organiss, areorganiss, are
responsible orresponsible or
6/? o inections6/? o inectionstreated in thetreated in the
developed world.developed world.
Biofls a Great threat toBiofls a Great threat to
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Biofls a Great threat toBiofls a Great threat to
:plants:plants - signifcant nuber o people are- signifcant nuber o people are
aAected by biofl inections whichaAected by biofl inections which
develop on edical devices iplanted indevelop on edical devices iplanted in
the body such as catheters !tubes used tothe body such as catheters !tubes used toconduct &uids in or out o the body,conduct &uids in or out o the body,
artifcial ;oints, and echanical heartartifcial ;oints, and echanical heart
valves. >hen iplanted aterialvalves. >hen iplanted aterial
becoes colonized by icroorganiss, abecoes colonized by icroorganiss, aslow developing but persistent inectionslow developing but persistent inection
results.results.
Biofls a Gr oing concern inBiofls a Grwoing concern in
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Biofls a Grwoing concern inBiofls a Grwoing concern in
odern edicineodern edicine
8rosthetic device inections, such as8rosthetic device inections, such as
those involving artifcial heart valves,those involving artifcial heart valves,
intravascular catheters, or prostheticintravascular catheters, or prosthetic
;oints, are prie e'aples o biofl%;oints, are prie e'aples o biofl%associated inections. >ith theassociated inections. >ith the
increasing use o such devices in theincreasing use o such devices in the
odern practice o edicine, theodern practice o edicine, theprevalence o these inections isprevalence o these inections is
e'pected to increase.e'pected to increase.
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Dental pla/ueDental pla/ue
ental pla(ue is aental pla(ue is a
yellowish bioflyellowish biofl
that build up onthat build up on
the teeth. : notthe teeth. : notreoved regularly,reoved regularly,
it can lead toit can lead to
dental caries.dental caries.
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Dental pla/uesDental pla/ues
#he oration o#he oration odental pla(ue biodental pla(ue bio
fls includes afls includes a
series o steps thatseries o steps that
begins with thebegins with theinitial colonizationinitial colonization
o the pellicle ando the pellicle and
ends with theends with the
cople' orationcople' oration
o a ature bio fl.o a ature bio fl.
'ormation of Dental'ormation of Dental
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o a o o e ao a o o e aBiolmsBiolms
-dditionally, through-dditionally, throughthe growth process othe growth process o
the pla(ue bio fl, thethe pla(ue bio fl, theicrobial copositionicrobial coposition
changes ro one thatchanges ro one thatis priarily gra%is priarily gra%
positive andpositive and
streptococcus%rich to astreptococcus%rich to astructure flled withstructure flled with
gra%negativegra%negativeanaerobes in its oreanaerobes in its ore
ature state.ature state.
e %ce s gna ng e'.e %ce s gna ng e'. /uorum/uorumii d i ti ith d i ti ith
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sensingsensing, and counication with, and counication with
diAerent bacteria enhance BiofldiAerent bacteria enhance Biofl
orationoration
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Bioflms everywhereBioflms everywhere
#heyKre everywhere9#heyKre everywhere9on your showeron your shower
curtain, on edicalcurtain, on edical
devices iplanted indevices iplanted in
patients, on roc)s inpatients, on roc)s inrivers and streas,rivers and streas,
and in your nose.and in your nose.
>hile the sheer>hile the sheer
nuber o diAerentnuber o diAerent
organiss a bioflorganiss a biofl
ay contain a)es itay contain a)es it
a challenge to study,a challenge to study,
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#D##D#77 on Bioflson Biofls
Biofls or on the surace oBiofls or on the surace o
catheter lines and contact lenses.catheter lines and contact lenses.
#hey grow on pacea)ers, heart#hey grow on pacea)ers, heart
valve replaceents, artifcial ;ointsvalve replaceents, artifcial ;ointsand other surgical iplants. #heand other surgical iplants. #he CDCCDC
!!enters for !isease ontrolenters for !isease ontrol
estiate that over 6/? oestiate that over 6/? o=osocoial !hospital%ac(uired=osocoial !hospital%ac(uired
inections are caused by biofls.inections are caused by biofls.
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Biofls interere in -ntibioticBiofls interere in -ntibiotic
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Biofls interere in -ntibioticBiofls interere in -ntibiotic
#herapy#herapy Bacteria growing in aBacteria growing in a
biofl are highlybiofl are highly
resistant to antibiotics,resistant to antibiotics,
up to 1,000 ties oreup to 1,000 ties ore
resistant than the saeresistant than the saebacteria not growing inbacteria not growing in
a biofl. *tandarda biofl. *tandard
antibiotic therapyantibiotic therapyisis
oten useless and theoten useless and the
only recourse ay beonly recourse ay beto reove theto reove the
containated iplant.containated iplant.
Biofl and -ntibioticBiofl and -ntibiotic
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Biofl and -ntibioticBiofl and -ntibiotic
resistanceresistance - )ey property o bio- )ey property o bio
fls is that individualfls is that individual
icroorganiss areicroorganiss are
bound together by abound together by a
polyeric substancepolyeric substancee'creted by thee'creted by the
icroorganiss.. #hisicroorganiss.. #his
protectiveprotective
encapsulation isencapsulation is
believed to play a rolebelieved to play a role
in soe antibiotic%in soe antibiotic%
resistant inection.resistant inection.
Bacterial resitance andBacterial resitance and
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Bacterial resitance andBacterial resitance and
BioflsBiofls -nother area o great iportance ro a-nother area o great iportance ro a
public health perspective is the role opublic health perspective is the role obiofls in antiicrobial%drug resistance.biofls in antiicrobial%drug resistance.Bacteria within biofls are intrinsicallyBacteria within biofls are intrinsicallyore resistant to antiicrobial agentsore resistant to antiicrobial agentsthan plan) tonic cells because o thethan plan) tonic cells because o thediinished rates o ass transport odiinished rates o ass transport oantiicrobial olecules to the bioflantiicrobial olecules to the biofl
associated cells or because biofl cellsassociated cells or because biofl cellsdiAer physiologically ro plan) tonicdiAer physiologically ro plan) toniccellscells
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Biolms in #ystic brosisBiolms in #ystic brosis
Biofls areBiofls areinvolved ininvolved in
nuerousnuerous
diseases. :n cysticdiseases. :n cystic
fbrosis patientsfbrosis patients
have 8seudoonashave 8seudoonas
inections thatinections that
oten result inoten result inantibiotic resistantantibiotic resistant
biofls.biofls.
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0ndocarditis and Biolms0ndocarditis and Biolms
icroorganiss ay attach and developicroorganiss ay attach and developbiofls on coponents o echanical heartbiofls on coponents o echanical heartvalves and surrounding tissues o the heart,valves and surrounding tissues o the heart,leading to a condition )nown as prostheticleading to a condition )nown as prosthetic
valve endocarditis. #he priary organissvalve endocarditis. #he priary organissresponsible or this condition areresponsible or this condition are ".".epidermidis, ". aureus, "treptococcusepidermidis, ". aureus, "treptococcus sppspp.,.,gra%negative bacilli, diphtheroids,gra%negative bacilli, diphtheroids,
enterococci, andenterococci, and
andidaandida
spp. #hesespp. #hese
organiss ay originate ro the s)in, otherorganiss ay originate ro the s)in, otherindwelling devices such as central venousindwelling devices such as central venouscatheters, or dental wor).catheters, or dental wor).
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0ye and Biolms0ye and Biolms
#he presence o#he presence obacterial biofls hasbacterial biofls hasbeen deonstrated onbeen deonstrated onany edical devicesany edical devicesincluding intravenousincluding intravenouscatheters, as well ascatheters, as well asaterials relevant to theaterials relevant to theeye such as contacteye such as contactlenses, scleral buc)les,lenses, scleral buc)les,suture aterial, andsuture aterial, andintraocular lenses. anyintraocular lenses. anyocular inections otenocular inections otenoccur when suchoccur when suchprosthetic devices coeprosthetic devices coein contact with or arein contact with or areiplanted in the eye.iplanted in the eye.
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Biofls and 2ontact lensesBiofls and 2ontact lenses
Bacterial bioflBacterial biofloration on contactoration on contactlenses and contact lenslenses and contact lensstorage cases ay bestorage cases ay bea ris) actor in contacta ris) actor in contact
lens%associated corneallens%associated cornealinections. *tudiesinections. *tudieshave shown thathave shown thatcontaination o lenscontaination o lenscases by bacteria,cases by bacteria,ungi, and aoebae isungi, and aoebae is
coon with 0? tocoon with 0? to0? o lens wearers0? o lens wearershaving a containatedhaving a containatedlens case.lens case.
rinary catheters andrinary catheters and
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rinary catheters andrinary catheters and
BiolmsBiolms Frinary catheters are tubular late' orFrinary catheters are tubular late' or
silicone devices, which when inserted aysilicone devices, which when inserted ayreadily ac(uire biofls on the inner orreadily ac(uire biofls on the inner orouter suraces. #he organiss coonlyouter suraces. #he organiss coonly
containating these devices andcontainating these devices anddeveloping biofls aredeveloping biofls are SSepidermidis! "nterococcus #aecalis! "epidermidis! "nterococcus #aecalis! "coli! Proteus mirabilis! P aeruginosa!coli! Proteus mirabilis! P aeruginosa!$ pneumoniae$ pneumoniae,, and other gra%negativeand other gra%negative
organiss. #he longer the urinary catheterorganiss. #he longer the urinary catheterreains in place, the greater the tendencyreains in place, the greater the tendencyo these organiss to develop biofls ando these organiss to develop biofls andresult in urinary tract inections.result in urinary tract inections.
Biofls and indwelling edicalBiofls and indwelling edical
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Biofls and indwelling edicalBiofls and indwelling edical
devicesdevices Biofls on indwelling edical devices ayBiofls on indwelling edical devices ay
be coposed o gra%positive or gra%be coposed o gra%positive or gra%
negative bacteria or yeasts. Bacterianegative bacteria or yeasts. Bacteria
coonly isolated ro these devicescoonly isolated ro these devices
include theinclude the gra%positivegra%positive #nterococcus#nterococcusfaecalis,faecalis,"taphylococcus aureus,"taphylococcus aureus,
"taphylococcus epidermidis"taphylococcus epidermidis, and, and
"treptococcus"treptococcus viridiansviridians$$ and the gra%and the gra%
negativenegative #scherichia coli, %lebsiella#scherichia coli, %lebsiellapneumoniae, &roteus mirabilis,pneumoniae, &roteus mirabilis, andand
&seudomonas aeruginosa&seudomonas aeruginosa..
%ndwelling catheters and%ndwelling catheters and
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%ndwelling catheters and%ndwelling catheters and
BioflmsBioflms&& 2entral venous catheters, the reerence2entral venous catheters, the reerence
ethod or (uantifcation o biofls onethod or (uantifcation o biofls oncatheter tips is the roll%plate techni(ue, incatheter tips is the roll%plate techni(ue, inwhich the tip o the catheter is reovedwhich the tip o the catheter is reoved
and rolled over the surace o aand rolled over the surace o anonselective ediu. Luantifcation o thenonselective ediu. Luantifcation o thebiofl depends on the nuber obiofl depends on the nuber oorganiss recovered by contact with theorganiss recovered by contact with theagar surace. Biofl%associated cells on theagar surace. Biofl%associated cells on the
inner luen o the device are not detectedinner luen o the device are not detectedwith this ethod, which has low diagnosticwith this ethod, which has low diagnosticsensitivity and low predictive value orsensitivity and low predictive value orcatheter%related bacteraeia.catheter%related bacteraeia.
:ndwelling catheters and:ndwelling catheters and
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:ndwelling catheters and:ndwelling catheters and
BioflsBioflsMM
:n addition, this ethod cannot detect:n addition, this ethod cannot detectore than 1,000 colony%oring unitsore than 1,000 colony%oring units!2$F per tip. - ethod that used!2$F per tip. - ethod that used
sonication plus vorte'ing as a eanssonication plus vorte'ing as a eanso (uantiying biofls on catheter tipso (uantiying biofls on catheter tipsshowed that a level o 10N 2$F per tipshowed that a level o 10N 2$F per tipis predictive o catheter%relatedis predictive o catheter%related
septicaeiasepticaeiaMMBiolms and Device(2ssociated InfectionsBiolms and Device(2ssociated Infections
3odney M. Donlan #enters for Disease #ontrol and 4revention3odney M. Donlan #enters for Disease #ontrol and 4revention2tlanta, 5eorgia, -22tlanta, 5eorgia, -2
-ntibiotic therapy alone ay-ntibiotic therapy alone ay
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-ntibiotic therapy alone ay-ntibiotic therapy alone ay
not cure Onot cure O -ntiicrobial agents are-ntiicrobial agents are
adinistered during valveadinistered during valvereplaceent andreplaceent andwhenever the patient haswhenever the patient hasdental wor) to preventdental wor) to preventinitial attachent byinitial attachent by)illing all icroorganiss)illing all icroorganissintroduced into theintroduced into thebloodstrea.bloodstrea. 2s ith2s ithbiolms on otherbiolms on otherindelling devices,indelling devices,
relatively fe patientsrelatively fe patientscan be cured of acan be cured of abiolm infection bybiolm infection byantibiotic therapy aloneantibiotic therapy alone
Biofls a concern inBiofls a concern in
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Biofls a concern inBiofls a concern in
-ntiicrobial #herapy-ntiicrobial #herapy icrobial biofls ay pose a public healthicrobial biofls ay pose a public health
proble or persons re(uiring indwellingproble or persons re(uiring indwelling
edical devices. #he icroorganiss inedical devices. #he icroorganiss in
biofls are diPcult or ipossible to treatbiofls are diPcult or ipossible to treat
with antiicrobial agentsI detachent rowith antiicrobial agentsI detachent rothe device ay result in inection. -lthoughthe device ay result in inection. -lthough
edical devices ay diAer widely in designedical devices ay diAer widely in design
and use characteristics, specifc actorsand use characteristics, specifc actors
deterine susceptibility o a device todeterine susceptibility o a device toicrobial containation and bioflicrobial containation and biofl
oration.oration.
Biofls need higherBiofls need higher
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Biofls need higherBiofls need higher
concentration o -ntibioticsconcentration o -ntibiotics Biofls are rear)ably diPcult to treatBiofls are rear)ably diPcult to treat
with antiicrobials. -ntiicrobials aywith antiicrobials. -ntiicrobials ay
be readily inactivated or ail to penetratebe readily inactivated or ail to penetrate
into the biofl. :n addition, bacteriainto the biofl. :n addition, bacteriawithin biofls have increased !up towithin biofls have increased !up to
1000%old higher resistance to1000%old higher resistance to
antiicrobial copounds, even thoughantiicrobial copounds, even though
these sae bacteria are sensitive tothese sae bacteria are sensitive tothese agents i grown under plan) tonicthese agents i grown under plan) tonic
conditions.conditions.
Biolms help 5eneBiolms help 5ene
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Biolms help 5eneBiolms help 5ene
transfertransfer Biofls increase theBiofls increase the
opportunity or geneopportunity or gene
transertranser
between
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Biolms uorumBiolms uorum
sensingsensing 2ertain species o2ertain species o
bacteria counicatebacteria counicatewith each other withinwith each other withinthe biofl. -s theirthe biofl. -s theirdensity increases, thedensity increases, theorganiss secrete loworganiss secrete lowolecular weightolecular weightolecules that signalolecules that signalwhen the populationwhen the populationhas reached a criticalhas reached a criticalthreshold. #his process,threshold. #his process,
calledcalled /uorum/uorumsensingsensing, is responsible, is responsibleor the e'pression oor the e'pression ovirulence actors.virulence actors.
uorum sensing helps theuorum sensing helps the
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uorum sensing helps theuorum sensing helps the
survival of pathogenssurvival of pathogens
Biolms contribute for neBiolms contribute for ne
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Biolms contribute for neBiolms contribute for ne
phenotypesphenotypes Bacteria e'press new, and soetiesBacteria e'press new, and soeties
ore virulent phenotypes when growingore virulent phenotypes when growing
within a biofl. *uch phenotypes aywithin a biofl. *uch phenotypes ay
not have been detected in the pastnot have been detected in the pastbecause the organiss were grown onbecause the organiss were grown on
rich nutrient edia under plan) tonicrich nutrient edia under plan) tonic
conditions. #he growth conditions areconditions. #he growth conditions are
(uite diAerent particularly in the depths(uite diAerent particularly in the depthso biofls,o biofls,
Biolms protects fromBiolms protects from
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Biolms protects fromBiolms protects from
Immune responsesImmune responses Bacteria ebedded within biofls areBacteria ebedded within biofls are
resistant to both iunological andresistant to both iunological and
non% specifc deence echaniss onon% specifc deence echaniss o
the body. 2ontact with a solid suracethe body. 2ontact with a solid suracetriggers the e'pression o a panel otriggers the e'pression o a panel o
bacterial enzyes which catalyze thebacterial enzyes which catalyze the
oration o stic)y polysaccharides thatoration o stic)y polysaccharides that
proote colonization and protection.proote colonization and protection.
Biolms 6 4rotects fromBiolms 6 4rotects from
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Biolms 4rotects fromBiolms 4rotects from
4hagocytosis4hagocytosis
8hagocytes are unable to eAectively8hagocytes are unable to eAectivelyengul a bacteriu growing within aengul a bacteriu growing within a
cople' polysaccharide atri'cople' polysaccharide atri'
attached to a solid surace. #hisattached to a solid surace. #hiscauses the phagocyte to releasecauses the phagocyte to release
large aounts o pro%in&aatorylarge aounts o pro%in&aatory
enzyes and cyto)ines, leading toenzyes and cyto)ines, leading toin&aation and destruction oin&aation and destruction o
nearby tissues.nearby tissues.
#urrent ob8ectives on#urrent ob8ectives on
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#urrent ob8ectives on#urrent ob8ectives on
Biolm researchBiolm research ooevelopent o iproved iaging oevelopent o iproved iaging o
biofls in situIbiofls in situIooevelopent o iproved clinicallyevelopent o iproved clinically
relevant in vitro and in vivo odels orelevant in vitro and in vivo odels o
biofls under specifc in vivo conditions suchbiofls under specifc in vivo conditions suchas &ow rate, nutrient content, andas &ow rate, nutrient content, andteperatureIteperatureI ooevelopent o better probes !genetic,evelopent o better probes !genetic,etabolic, and iunological or real% tieetabolic, and iunological or real% tieanalysisIanalysisI oo*tudies o (uoru sensing
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2urrent ob;ectives on Biofl2u e ob;ec es o o
researchresearch oo $urther characterization o biofl%specifc$urther characterization o biofl%specifc
gene e'pressionIgene e'pressionI oo*tudies o the e'change o genetic aterial*tudies o the e'change o genetic aterial
within bioflsIwithin bioflsI oo
*tudies o organic containants on substrata,*tudies o organic containants on substrata,and their in&uence on biofl structureIand their in&uence on biofl structureI
oo evelopentevelopent
o novel approaches to control pathogenico novel approaches to control pathogenic
bacteria by, or e'aple, devising strategiesbacteria by, or e'aple, devising strategiesto avour growth o non%pathogenicto avour growth o non%pathogenic
icroorganiss in biofl counitiesIicroorganiss in biofl counitiesI
2urrent ob;ectives on Biofl2urrent ob;ectives on Biofl
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2urrent ob;ectives on Biofl;
researchresearch oo*tudies o pathogenic*tudies o pathogenic
echaniss o icrobesechaniss o icrobes
growing in bioflsIgrowing in bioflsI
oo@lucidation o@lucidation o
echaniss oechaniss o
resistance o biofls toresistance o biofls to
antiicrobial agentsIantiicrobial agentsI
oo-tudies of-tudies of
host immunehost immune
responses, bothresponses, bothinnate and adaptiveinnate and adaptive
to biolmsto biolmsII
2urrent ob;ectives on Biofl2urrent ob;ectives on Biofl
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2urrent ob;ectives on Biofl;
researchresearch :n studies o inectious lung disease in cystic:n studies o inectious lung disease in cystic
fbrosisIfbrosisIoo*tudies on the potential o diagnostic*tudies on the potential o diagnosticprocedures such as Bronchoalveloar lavageprocedures such as Bronchoalveloar lavageand bronchoscopy to disturb local biofland bronchoscopy to disturb local biofl&ora and inoculate distant locationsI&ora and inoculate distant locationsI
ooevelopent oevelopent oatheatical odels and coputeratheatical odels and coputersiulations o bioflsIsiulations o bioflsI
oo evelopent o theevelopent o the
ethodology or the prevention and controlethodology or the prevention and controlo biofls ro catheters, water unit lines,o biofls ro catheters, water unit lines,and other clinically iportant solid suracesI.and other clinically iportant solid suracesI.
*earching or alternatives*earching or alternatives77
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*ea c g o a e a esg
#issue engineering#issue engineering Hole o biofls in ultiple pathologies and theHole o biofls in ultiple pathologies and the
diPculty in resolving these pathologies spea)sdiPculty in resolving these pathologies spea)s
to the iportance o developing eans oto the iportance o developing eans o
replacing or enhancing the therapies alreadyreplacing or enhancing the therapies already
in use. #he use o synthetic aterials in thein use. #he use o synthetic aterials in thebody ranges ro catheters to esh to stentsbody ranges ro catheters to esh to stents
to heart valves and beyond. Fntil theto heart valves and beyond. Fntil the
developent o viable and practical tissuedevelopent o viable and practical tissue
engineering, then nuber and types oengineering, then nuber and types oapplications in which synthetic aterials areapplications in which synthetic aterials are
used will continue to increase.used will continue to increase.
0 i M th d0merging Methods
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0merging Methods0merging Methods
*everal researchers*everal researchersare fnding solutionsare fnding solutions
or the cure o Bioflsor the cure o Biofls
, yet it is e'periental,, yet it is e'periental,
with advances inwith advances inolecular biologyolecular biology
better odelbetter odel
treatents can betreatents can be
identifed to reduceidentifed to reduce
the proble o Bioflthe proble o Biofl
intererence inintererence in
-ntibiotic therapy.-ntibiotic therapy.
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2reated or2reated or r.#.J.Hao Qsr.#.J.Hao QsReQReQ
learning series.learning series.
@ail@ail
doctortvrao9gmail.comdoctortvrao9gmail.com
mailto:[email protected]?subject=Biofilmmailto:[email protected]?subject=Biofilmmailto:[email protected]?subject=Biofilm -
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@ndodontic biofls could be classifed as ollows9 1S@ndodontic biofls could be classifed as ollows9 1S:ntracanal biofls that are generally present on the:ntracanal biofls that are generally present on the
root canal dentine o an endodontically inected tooth.root canal dentine o an endodontically inected tooth.
S @'traradicular icrobial biofls that are ored onS @'traradicular icrobial biofls that are ored on
the root !ceentu surace ad;acent to the ape' othe root !ceentu surace ad;acent to the ape' o
endodontically inected teeth. 3S 8eriapical icrobialendodontically inected teeth. 3S 8eriapical icrobialbiofls that are isolated biofls ound in the periapicalbiofls that are isolated biofls ound in the periapical
region o an endodontically treated tooth. NS Bioaterialregion o an endodontically treated tooth. NS Bioaterial
centered inection that is ored when bacteria adherescentered inection that is ored when bacteria adheres
to an artifcial bioaterial surace such as root canalto an artifcial bioaterial surace such as root canal
obturating aterials, thereby oring a biofl. #heseobturating aterials, thereby oring a biofl. #hesebiofls could be intraradicular or e'traradicular,biofls could be intraradicular or e'traradicular,
depending on the apical e'tent o the obturation.3depending on the apical e'tent o the obturation.3
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@ndodontic icro&ora9@ndodontic icro&ora9 ost o the bacteria in an endodontic inection areost o the bacteria in an endodontic inection are
strict anaerobes. #he &ora is usually polyicrobial,strict anaerobes. #he &ora is usually polyicrobial,
doinated by obligate anaerobic bacteria. Gra%doinated by obligate anaerobic bacteria. Gra%
positive organiss are ound in appro'iately /?positive organiss are ound in appro'iately /?o the saplesI the ost predoinant areo the saplesI the ost predoinant are
streptococci !?, staphylococci !1/?,streptococci !?, staphylococci !1/?,
corynebacteria !10%/?, yeasts !1?, and others.corynebacteria !10%/?, yeasts !1?, and others.
#he Gra%negative bacteria !N? include#he Gra%negative bacteria !N? include
spirochetes !5%1?, neisseria !N?, bacteroidesspirochetes !5%1?, neisseria !N?, bacteroides
!?, usobacteriu !3?, pseudoonas !?,!?, usobacteriu !3?, pseudoonas !?,
colior bacteria !1?, and otherscolior bacteria !1?, and others
#ypes o @ndodontic#ypes o @ndodontic
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ypyp
:nections:nections :S :ntraradicular :nection:S :ntraradicular :nection 8riary intraradicular inections98riary intraradicular inections9
icroorganiss that initially invade and colonize theicroorganiss that initially invade and colonize the
necrotic pulp tissue cause priary intraradicularnecrotic pulp tissue cause priary intraradicular
inection. 8riary inections are characterized by ainection. 8riary inections are characterized by ai'ed consortiu coposed o 10 to 30 bacteriali'ed consortiu coposed o 10 to 30 bacterial
species and 103 to 10 bacterial cells per canal. #hespecies and 103 to 10 bacterial cells per canal. #he
involved icrobiota is conspicuously doinated byinvolved icrobiota is conspicuously doinated by
anaerobic bacteria, but soe acultative oranaerobic bacteria, but soe acultative or
icroaerophilic species can also be coonly oundicroaerophilic species can also be coonly ound
in priary intraradicular inections.Nin priary intraradicular inections.N
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*econdary :ntraradicular :nections9*econdary :ntraradicular :nections9 icroorganiss that were not present in theicroorganiss that were not present in the
priary inection but that were introducedpriary inection but that were introduced
into the root canal syste at soe tie aterinto the root canal syste at soe tie ater
proessional intervention cause secondaryproessional intervention cause secondary
intraradicular inections. #he entry can beintraradicular inections. #he entry can be
during treatent, between appointents, orduring treatent, between appointents, or
even ater root canal flling. *pecies involvedeven ater root canal flling. *pecies involved
can be oral or nonoral icroorganiss,can be oral or nonoral icroorganiss,
depending on the cause o inection.Ndepending on the cause o inection.N
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8ersistent :ntraradicular :nection9icroorganiss that can resist
intracanal antiicrobial proceduresand endure periods o nutrientdeprivation in a prepared canal causepersistent intraradicular inections.#his is also tered recurrentinection. :nvolved icroorganiss
are renants o a priary orsecondary inection. @ aecalis arepredoinant and is a persistentorganis. :t is coonly ound in ahigh percentage o root canal as asingle organis or as a a;orcoponent o the &ora. 8ersistent
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::S @'traradicular :nection9icrobial invasion o the in&aedperiradicular tissue is invariably ase(uel o interradicular inection.-cute alveolar abscess is an e'apleo e'traradicular e'tension or a
se(uel to interradicular inection.*oeties e'traradicular inectioncan be independent o intraradicularinections. $or e'aple, apicalactinoycosis caused by-ctinoyces species and 8.
propionicu is a pathological diseasewhich can be treated only byperiapical surgery.
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-trategies to prevent biolm formation depend on conte!t-trategies to prevent biolm formation depend on conte!t I2: :I;03-I2: :I;03-
gppgpp
Medical "small scale&Medical "small scale&
-I*< =' >I'0-I*< =' >I'0
#hemical approaches?antibiotics, biocides etc.#hemical approaches?antibiotics, biocides etc.
4hysicalapproaches?seofacousticenergyultrasoundvibration4hysicalapproaches?seofacousticenergyultrasoundvibration
-#I0:#0--#I0:#0- 4hysical approaches?se of acoustic energy, ultrasound, vibration, 4ie@o electric4hysical approaches?se of acoustic energy, ultrasound, vibration, 4ie@o electric
elements Biological approaches?>ytic bacteriophages, interference ithelements Biological approaches?>ytic bacteriophages, interference ith
Idtil"ll&Idtil"ll&
inter(bacterial signaling, interference ith genetic programs that control biolminter(bacterial signaling, interference ith genetic programs that control biolm
formationformation
Industrial "large scale& #hemical approaches?chlorine, o@one, ioni@ation, hydrogenIndustrial "large scale& #hemical approaches?chlorine, o@one, ioni@ation, hydrogen
pero!ide, various biocidespero!ide, various biocides 4hysical approaches?choice of material that inhibits attachment, vibration,4hysical approaches?choice of material that inhibits attachment, vibration,
mechanical cleaning "e.g. high pressure )ushing&, ;mechanical cleaning "e.g. high pressure )ushing&, ;
.umb.no.umb.no
Biological approaches?alteration of groth conditionsA,Biological approaches?alteration of groth conditionsA,
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ethods or analysing icrobial biofls9ethods or analysing icrobial biofls9 :-= F=:J@H*:-= F=:J@H*
ygyg
@lectron icroscopy@lectron icroscopy
*:#T C$ :$@*:#T C$ :$@
=anoscale iaging2onocal laser scanning icroscopy @pi&uorescence=anoscale iaging2onocal laser scanning icroscopy @pi&uorescence
icroscopy -toic orce icroscopyicroscopy -toic orce icroscopy
*2:@=2@**2:@=2@*
=anoscale analysis=anoscale analysis
#ip enhanced Haan spectroscopy#ip enhanced Haan spectroscopy
ass spectroscopyass spectroscopy
llthillthi icroarray analysis olecular techni(ues>hole genoe se(uencingicroarray analysis olecular techni(ues>hole genoe se(uencing
$luorescent labeling techni(ues$luorescent labeling techni(ues
@ssential )now howicrobial physiology and etabolis icrobial genetics@ssential )now howicrobial physiology and etabolis icrobial genetics
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8ossibly the frst identifcation o8ossibly the frst identifcation obiofl structures in inected rootbiofl structures in inected root
canals was carried out by =air !10.canals was carried out by =air !10.
Cn the basis o transission electronCn the basis o transission electronicroscopy !#@ he e'ained theicroscopy !#@ he e'ained the
root canal content o 31 teeth, whichroot canal content o 31 teeth, which
had gross coronal caries and to whichhad gross coronal caries and to whichthe periapical in&aatory processthe periapical in&aatory process
was attached upon e'tractionwas attached upon e'traction
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he bacterial condensations showed ahe bacterial condensations showed apalisade structure siilar to the onepalisade structure siilar to the one
or dental pla(ue on e'ternal toothor dental pla(ue on e'ternal tooth
suraces, suggesting siilarsuraces, suggesting siilarechaniss or bacterial attachentechaniss or bacterial attachent
as those or dental pla(ueas those or dental pla(ue
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olven et al. !11 noted by scanningelectron icroscopic !*@observations o the apical oinected root canals that cocci androds and
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. :n a study o cases resisting treatent, the so%. :n a study o cases resisting treatent, the so%called reractory endodontic cases, #ronstad et al.called reractory endodontic cases, #ronstad et al.
!N e'ained the suraces o the root tips reoved!N e'ained the suraces o the root tips reoved
during surgical intervention by *@. #hey notedduring surgical intervention by *@. #hey noted
that the ape' o the roots ad;acent to the apicalthat the ape' o the roots ad;acent to the apicaloraen was coated with a continuous, sooth,oraen was coated with a continuous, sooth,
structure%less layer containing a variety ostructure%less layer containing a variety o
bacterial ors. :n irregularities o the suraces andbacterial ors. :n irregularities o the suraces and
in crypts and holes, bacteria were seen heldin crypts and holes, bacteria were seen held
together by an e'tracellular aterial. #hetogether by an e'tracellular aterial. #heorganiss were identifed as cocci and rods withorganiss were identifed as cocci and rods with
soe presence o fbrillar orsoe presence o fbrillar or
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=oiri et al. ! analyzed the presence o=oiri et al. ! analyzed the presence obiofl orations on root tips o e'tractedbiofl orations on root tips o e'tracted
teeth with Rreractory periapical pathosisQteeth with Rreractory periapical pathosisQ
and gutta%percha points reoved duringand gutta%percha points reoved during
endodontic treatent by *@. Guttaperchaendodontic treatent by *@. Guttapercha
points stic)ing out through the ape' werepoints stic)ing out through the ape' were
alost copletely covered with glycocaly'%alost copletely covered with glycocaly'%
li)e structures. Bacteria, ostly flaentsli)e structures. Bacteria, ostly flaents
or long rods, were seen on the e'ternal rootor long rods, were seen on the e'ternal root
suraces in the e'tracted teethsuraces in the e'tracted teeth