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Clostridia
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Clostridia
LargeGrampositiveanaerobicrods
Formofsporesresistanttooxygen,drought,
freezingandcookingtemperatures
Thesoilandintheintestinesofwarmblooded
Causethefood,theenvironmentandthepointsof
throughdrugpoisoningandinfectious
thetoxininfections
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A.Clostridiumbotulinum
1.Theclassicbotulism
2.Theinfantbotulism
3.Thewoundbotulism
B.Clostridiumperfringens
1.Theclassicfoodpoisoning
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2.enteritisnecroticans
C.clostridiumtetani
1.Tetanus
D.Clostridiumdifficile
1.Clostridiumdifficileassossioitunutdiarrhea(CDAD)
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A.Clostridiumbotulinum
1.Classicalbotulism
Foodpoisoning,theriskoffoodproducts,especiallymeat,fish,vacuumpacked
Productsandpreservedhomeproducts
Clostridiumbotulinumsecretesproteiinieksotoksiinia,whichisthestrongest
knownastoxic(lethaldose:0.1ng/kg)
Neurotoxinsaresynthesizedasaninactivepolypeptide,i.e.,
protoksiineina(size150kDa),whichareactivatedwhentheproteasecleaves
polypeptidechain
C.botulinumareclassifiedintosevenserotypesAGduringgrowth
onthebasisoftheantigeniccharacteristicsoftheirproductionoftoxinstypesA,
B,EandFcausebotulismpeople
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Afewotherklostridilajit(C.butyricum,C.Barati)toproduce
thesametoxinsasC.Botulinum
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Functionalformofbotulinumtoxincanbind
nervemusclejointsandpreventstheneurotransmitter
releaseofvesiclescontainingacetylcholine.Since
themusclemaynolongerbeanykindofmessagestothenervecells,muscle
paralyzed.
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thetoxinisabsorbedinthestomachandsmallintestine,fromwhereit
spreadsthroughoutthebody,canalsobeabsorbedbyinhalation
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Symptomsbegin1236hoursafteringestionoffood
Thefirstbotulismsymptomsincludefatigue,weakness,
vomiting,thenvoiceanddifficultyinswallowingandvisualdisturbances
Respiratorydepressioncanultimatelyleadtodeath
Mortalityofthediseaseisabout65%,indevelopedcountriesis10%..
Riskgroup:dangerousforeveryone
Evenoneperson'sillnessbotulismepidemicisinterpreted
Duetothenatureofthe
aseriousdisease
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2009
InearlySeptember,itwasfoundinFranceinthreeatthesame
thefamilyofapersonwhoisClostridiumbotulinum
botulismcausedbybacteria,thesuspectedsource
PurchasedfromFinlandtohotsmokedwhitefish.Vacuumpackedfish
waspurchasedinEasternFinlandinretailtradeinAugust
Attheend,transportedbyplanetoFrance
andbyaprivateindividualeatentwoweeks
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lateronthelastdayofuse.Journey,thefish
hadbeenstoredinacoolbox
unknowntemperaturefor14hoursandthen
Afterthehomerefrigerator.Beforeeatingfishisnot
washeated.
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2006
ClostridiumbotulinumcausedinJuly2006forone
epidemic,whichcontractedbytwopersonsofthesame
family.Thevehicleswere
likelytobehotsmokedwhitefish,whichwaspurchased
packedinalocalsupermarket.Fishfinal
datewasweeksawaysymisajankohdasta.Fish
thecountryoforiginwasCanada,itwasdeliveredtoFinland
frozen.Defrostingandsmoking,aswellasthefinishedproduct
packagingtookplaceinadomesticestablishment.
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Olderepidemics
1981twoGermantouristsgotsickaftereating
silykemaksapasteijaa
1999onepersonillwithwhitefishroe
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Botulismtreatment:
ABEtrivalentantitoxin(givenbeforesymptoms
thestart)
Gastrointestinaltractiscleared
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Missedthegutbacteriaaredestroyedpenicillin
Artificialrespiration
Forecasttheworsethesoonerthesymptomsappear
Recoverymaytakemonths
Waitingfornewnervemusclejunctionsformation
Smalldosesofpurifiedbotulinumtoxincanbe
useofCNSlihastondysregulation,
inthetreatmentofdiseases,thecosmeticuseofbiological
warfare
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2.Theinfantbotulism
C.botulinumproducestoxininthebaby'sgastrointestinaltract(n.80
spores/g)
Riskfoodshoney,vegetablesandfruits
Lessthanhalfyearsmorelikelytogetinfantbotulism
immatureintestinalflora,andahigherpHinthedigestivetract
allowbacterialsporegrowthandyieldofthetoxininthegut
Symptomsofconstipation,muscleslackening,weepingsluggish
Thesymptomsaresurprisinglymild,andmortalityonlyafewpercent,
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causingsuddeninfantdeathsyndrome
Poweroftreatmentisusuallyrescued
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3.Thewoundbotulism
C.botulinumspore'swound,wheretheygerminate
Thediseaseresemblestetanus
Theuseofinjectingdrugs
CasesdetectedintheAmericas,Europe,rare
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Clostridiumbotulinum
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taskanalysisinmice
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Clostridiumbotulinum
PCR/PFGE/ELISA
M.LindstrmFinland,studiedthemethod:
kytettnprimeriparia4,whichare
specificneurotoxinsbotulinumtoxinA,B,Eor
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F.Fourserotypesimultaneousdetection
possible.
ELISAreadyforadaywhenthetoxininthesample
Aviablecellgrowthandtoxin
generatingidentificationofgenesbyPCR
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B.Clostridiumperfringens
1.Theclassicfoodpoisoning
C.perfringensTypeAenterotoxinsecretedlast
years,themostcommonlyreportedcauseoffoodpoisoning
InFinland
Theriskoffoodmeatandfish,incompletelycooked,
slowlychilled,attoohighatemperature,storedand
incompletelycookedfoods
Causingdiseasedoseof10^6to10^8cells
Theincubationperiodof824h
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C.perfringenscellsincreaseinfood,someofthecellsdieofthestomach
acidity,butsomecellsremainaliveinthesmallintestine,where
theybegintoproliferate
Whenthebacterialcellitii,consistspoisoningcausingCPE
enterotoxin,ifthepositioniscpegeneencodingit(only5%
strains)
CPEtoxindamagethesurfaceoftheepithelialcellsofthesmallintestine
bymakingholesinthecellmembranestosmallmoleculessuchasanions,
cationsandorganicacidsleakout
Symptomsofabdominalpain,nauseaandintensediarrheathatlastfor1to2
days,theelderlyC.perfingenscancausepersistentdiarrhea
Enterotoxincanbedemonstratedinfaeces
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2.Enteritisnecroticans(pigbel)
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ClostridiumperfingensTypeCsecretedby
enterotoxincancausesevereenteritis
necroticansin(necrotizinginflammationofthebowel)
Thebacteriasecreteatoxinfromthefood,which
leadtoparalysis,haematomasandregional
necrosis,especiallyinthesmallintestine
Symptoms:vomiting,diarrhea,abdominalpainHard,
hemorrhage
Rare
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Clostridiumperfringens
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Clostridiumperfringensplating
TSCAgar(TryptoseSulfite
CycloserineAgar)
toproduceblackcolonies
cycloserinetopreventotherbacteria
growth
Inadditiontokanamycinandpolymyxin
inhibitionofbacterialflora
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TSC
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ClostridiumperfringensPCRandmicroarray
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Microarraytechnologywithantibodies
IdentificationofC.perfingenstoxins
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C.Clostridiumtetani
1.Tetanus,orlockjaw
C.tetaniproducestwopoison,andtetanoslysiinia
tetanospasmiinia,ofwhichthelatterissaidto
tetanustoxin
Tetanusneurotoxin,andthesecondstrongestpoison
Afterbotulinumtoxin
Thebacteriacancomeintothewound,whereitbeginstoincrease
underanaerobicconditions
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Infectionislocal,butthetoxinspreads
viathebloodstreamandlymphcirculation,aswellasdirectlytothe
thenervefibersalongthespinalcordandbrainstem
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toxinaffectsthecentralnervoussystem
hermosynapseihinandmotor
endplates,blockstheinhibitory
neurotransmitters(e.g.glycine)
release,whereinthemuscle
ispreventedandtherelaxationofthe
musclespasms
Theincubationperiodofdaystomonths
Atthebeginningofmusclespasmsofthewound
close,spreadstothemusclesofthejawand
allpartsofthevoluntarymuscles
Tetanusleads
ifuntreated,isalmostalways
death,manicuredmortality
1030%
Preventedthroughvaccination
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Clostridiumtetani
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C.tetanipenetration
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Clostridiumtetanidetermination
GeneticallyIgGELISAIgGclassaredetermined
antibodies
MouseTest
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Clostridiumtetani,theMousetest
Thelefthindlegofthemouseinjected
Tetanustoxin
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Clostridiumdifficile(40006000cases/yr)
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InsulationCDS(C.difficile
SELECTIVE)agarcontainingaDcycloseriniaand
Cefoksitinia
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Clostridiumdifficile
Clostridiumdifficileassociateddiarrhea
(CDAD)
DiagnostictestsforCDAD:in
thedeterminationofthemarket.They
oftoxinB(CytoToxassay)
ortoxinsAandB(immunoassay)
thegeneticanalysis.
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QuickTest:ClostridiumdifficileToxinAdetection
ofC.difficiletoxinAinfaeceswith
Rapydtest
1015min.
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Sources:
Austin,JWandDoddsKL1997.Clostridiumbotulinum.In:FoodMicrobiology.ASMPress.Pages
288304.
Hielm,S.clostridia,thequiet.In:M.SalkinojaSalonen(ed.).Microbiologicalcriteria.Microbiology
publications49/2000.
MicrobiologyDepartmentofAppliedChemistryandMicrobiology,UniversityofHelsinki.Pages652
655.
SagittariusSomer,H.andRistola,M.2003.Clostridiumspecies.In:P.Huovinen(ed.).
Microbiologyand
infectiousdiseases.Book1.DuodecimMedicalPublications.Pages228236.
Madigan,MT2003.BrockBiologyofMicroorganism.PearsonEducational.
Guidemicrobiologicalfoodanddrinkingwaterhazards.EvidentialEELAOpas1/2003.Pages3744.
McClaneBAin1997.Clostridiumperfringens.In:FoodMicrobiology.ASMPress.Pages305326.
SalkinojaSalonen,M.clostridialtoxins.In:M.SalkinojaSalonen(ed.).Microbiologicalcriteria.
Microbiologypublications49/2000.
DivisionofMicrobiology,DepartmentofAppliedChemistryandMicrobiology,Helsinki
University.Pages548549.
Pictures:
http://www.agen.ufl.edu/chyn/age2062/lect/lect_25/FG19_012.GIF
http://wwwuser.gwdg.de/hbruegg/ct/tetanus2.gif
http://www.agen.ufl.edu/chyn/age2062/lect/lect_25/FG19_011.GIF
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Fountains
Lindstrm,MiiaKeto,RiikkaMarkkula,AnnukkaNevas,MariHielm,
SebastianKorkeala,Hannu.MultiplexPCRAssayforDetectionand
IdentificationofClostridiumbotulinumtoxinTypesA,B,E,and...&Applied
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EnvironmentalMicrobiology,Dec2001,Vol.67Issue12,p5694,6p,4
charts,1bw(AN6063605)
http://www.cfsan.fda.gov/ebam/bam17.html
www.google.fi
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