Bacilli in Food-Eng

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Bacilliinfood
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Bacilliinfood
MICROFoodandEnvironmentalHygieneandcontrol
2011November21
MirjaSalkinojaSalonen
SalkinojaSalonen,M.,Bacillus
foodstuffs2011November21
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CausingfoodpoisoningBacillusismainlyintwogroups:
Bacilluscereussensibarn(includesspeciesofB.cereussensustricto,B.thuringiensis,B.
mycoides,B.pseudomycoidesB.weihenstephanensis,B.anthracis)
and
Bacillussubtilisgroup(includingB.subtilis,B.amyloliquefaciens,B.mojavensisB.
atrophaeus).
Ofthese,theFinnishlawotherbiologicalriskgroups,...
17.10.2000ENOfficialJournaloftheEuropeanCommunitiesL262/21
DIRECTIVE2000/54/ECOFTHEEUROPEANPARLIAMENTANDOFTHECOUNCIL
of18September2000,
https://translate.googleusercontent.com/translate_f
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theprotectionofworkersfromtherisksrelatedtobiologicalagents
atwork
(SeventhreferredtoinArticle16ofDirective89/391/EEC,Article1
individualdirective
...Otherthan#1,isonlyB.anthracis(riskcategory3).InsomeEUcountries
B.cereusisariskgroup2,butthusFinland.B.anthracisdoesnotcause
foodpoisoning,butitssporescauseadangerousinfection
inhaledorthroughtheskin.
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Table10.IntrinsicandextrinsicpropertiesofB.cereusandB.subtlisgroupnoticeforgrowthin
foods
Bacterialbiochemicalproperties
starchdegradingactivity
AbilitytohydrolyzestarchisacommonpropertyofB.cereusgroup
species 1 andBacillusisthemostnoticeamylolyticgenusfor
2.Most
Industrialproductionofdifferentstarchdegradingenzymes
emeticstrainsarenegativeforstarchHydrolysisalthoughtheyare
mostfrequentlytoreportedinstarchyfoods.
proteaseactivity
noticeformakingaminoacidsavailablefromproteinSubstrates
lipolyticactivity
noticeformakingfattyacidsavailablefromlipidSubstrates
salttolerance
Allowssurvivalinsaltstressenvironment.ImportantAlsoforthemotocross
protectionbetweenstresses.ForexampleinonestudyisB.cereus,
saltprotectedagainsthydrogenperoxide,Whichprotectedagainst
3
ethanol,Whichprotectedagainstheatshock.
4
Intrinsicparametersofthefood
pH
B.cereusandB.subtilisgroupgrowatapHof5to9.5.Thepresenceof
saltaprwidenthepHrange.ToolowortoohighpHalterthe
Functioningofenzymesandthetransportofnutrientsintothecell.
Moisturecontent
thewateractivity(a
w)Ofmostfreshfoodsisabove0.99andB.
subtilisforexamplecangrowatminimum0.95.Theminimuma
w
5.
toreportedforB.cereusgrowthis0.91to0.96infriedrice
Temperatureandnutrientsmightpermitgrowthatlowervalues
ofa
w:
therangeofa
w OverWhichgrowthoccursisgreatestattheoptimum
temperatureforgrowthandthepresenceofnutrientsincreasesthe
rangeofa w OverWhichtheorganismscangrow.Alowa
w increases
lengthofthelagphaseofgrowthanddecreasesthegrowthrate.
oxidationreduction
GenerallythemembersofthegenusBacilluspreferapositiveEh
PotentialEh
values
(oxidized)forgrowth.
nutrientcontent
includesthepresenceofwater,sourceofenergy(suchassugarsand
aminoacids),sourceofnitrogen(likeaminoacids),vitaminsand
relatedgrowthfactorsandminerals.
Antimicrobialconstituents
Includesessentialoilsinplants(likeEugenolincloves,allicinin
Preservativesandfood
garlic,thymolinsageandoregano,etc),Lysozymeineggsandmilk,
Lactoferrininmilk,etc.FoodPreservativesaffectingsporeFORMERS
includenisin,benzoate,sorbate.
biologicalstructure
thenaturalcoveringofsomefoodsprovidesprotectionagainstthe
entryofspoilageorganisms(ietestaofseeds,shellsofeggs,ofnuts,
theskin,etc).Presenceoforganicmatteraprprotectbacteriafrom
theactionofdisinfectants.
4
Extrinsicparameters
temperatureofstorage
Generally,atemperaturelowerthan47Corhigherthan55Cwould
impedethegrowthofanymemberfromB.cereusorB.subtilisgroup.
(Table1andTable6).
relativehumidityof
Whenthea w ofafoodisforexample0.6,itisnoticetostoreit
environment
underconditionsThatPreventthefoodabsorbingmoisture.
thepresenceandconcentration
CO SalkinojaSalonen,M.,Bacillus
2 isthemostnoticeatmosphericgasThatisusedtocontrol
ofgases
microorganismsinfoods.Bacillussp.areamongthemostsensitive
microorganismstoCO
2 relativetomodifiedatmospherepackaging
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B.cereusisacommonbacterialeverywhereintheenvironment,especiallyplants(including
crops)intheroot(rhizosphere)andalsoplantendophyte.B.cereusisnot
harmfultoplantsandisfoundintubersandgreenparts,includingwoody.
Plantsarenotcontainedsterile(withtheexceptionoffruit)asanimals.Plant
watercanalsfoundinbacteriathatshallcomeintosoilwaterwiththerootsystem.
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Table2.EnvironmentalnichesofB.cereusgroup
soil a
Species
water b
food c
mammalian
gut d
+s
insect
gut e
+
+
B.cereussensustricto+
+
+
B.anthracis
+
B.thurigiensis
+
+
+
+s
B.mycoides
+
+
+
B.pseudomycoides
+
B.weihenstephanensis+
+
a Vilainetal.,2006Jensenetal.,2003vonStettenetal.,1999Thorsenetal.,2006
earthworm
gut f
plant
rhizosphere
or
endophyte
+
+
+
+
+
+
+
+
+
+
+
b stensviketetal,2004..Jensenetal,2003
c AstraightforwardArnesenetal.,2008Povetal.,2003Kajikazawaetal.,2007Frederiksenetal.
2006Hansonetal.,2005
d Wilksetal.,2007Jensenetal.,2003
e Jensenetal.,2003Cooketal.,2007
f Jensenetal.,2003
g Jensenetal.,2003Okunishietal.,2005SaileandKoehler,2006Jafraetal.,2006
s transient
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Table2.ThepublishedcompletegenomesequencesofB.cereusstrains.
Numberof
genes
BacilluscereusstrainSize(bp)
Links
Reference
ATCC10987
5224283
5603
NCBIRefseqNC003909Raskoetal.,2004
ATCC14579
5411809
5234
NCBIRefseqNC004722Ivanovaetal.,2003
5300915
5134
4087024
4165
NCBIRefseqNC009674
Unpublished
5300915
5269
NCBIRefseqNC006274
Hanetal.,2006
ZK
NVH39198
E33L
TIGR,Tax288681
Hanetal.,2006
Bacilluscereusisthe2000'sintensiveresearchastrack,becausethe
ruokamyrkyttjnhasincreasedtheimportanceoflongtermfoodlogisticschains
timeandnumberofmoveseatingcomfortfoods(heatandchill"kitchens").B.
cereuPRINCIsporesarehighlyresistantkuumuttaandaccumulateinthefood
productionprocesses,withtheheatingstep(=rivalsarereduced).
Formorethan10B.theentiregenomescereusstrainissequenced.
B.cereusandB.somestockssubtilisgroupproducingThermotolerantpeptiditoksiineja,
whicharehighlytoxictohumans,andhavecausedkuoleemankinled
foodpoisoning.Thesetoxinsremaininthefood,eventhoughitisbyheating
destroyedalllivingbacteria.
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Bacilluscereusisaveryheterogeneousgroup,eventhoughitsmembersseparationofthe
morphologicalorbiochemicalcharacteristicsisdifficult.Thistablecontainsteststhatcan
usedtodistinguishpathogenic(sub)speciesfromtheother.
GammafaagiapidetnasspecificB.anthracisspeciesof,butgammafaagilleherkkiB.cereusstrains
ismm.foundinthedairies'koticereuksista".TheyarenotexpectedtoproducehavattuB.anthracis
characterizedtoxins(lethaltoxinandedematoxin).B.anthracisthespeciesmostresemble
oksetustoksiinia(cereulide)producedbytheB.cereusstrainsthatarenegative(oralmost
negative)haemolysisasB.anthracis.
.
B.mycoides,andB.pseudomycoidesareknowntohavecausedfoodpoisoning
PhenotypictraitsusefultodifferentiatewithinthemembersofB.cereusgroup
Species
B.cereussensustricto+
B.anthracis
B.thurigiensis
B.mycoides
B.pseudomycoides*
B.weihenstephanensis+
haemolysismotilityparasporal
inclusion
+
+
+
+
+
susceptibility
topenicillin
colony
morphology
white
white
white/gray
rhizoid
white/cream,
rhizoidal
white
growth
temperature(C)
455
1540
1045
1040
1540
lysis
by
gamma
phage
738
*DistinguishedfromB.mycoidesbydifferencesinwholecellfattyacidcomposition12:0isoand13:0
anteisolevelsandfromB.cereusbydifferencesin12:0iso,12:0,15:0isoand16:0(Nakamura,1998).
4of5strainsofB.mycoideswereresistantand5of6strainsofB.pseudomycoideswereSusceptible
DatafromLunaetal.,2007
CompiledfromGranum,2007Gordon,1973Lechneret.al.,1998Nakamura,1998vonStettenetal.
1999Roberts,etal.,1996b.
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BiochemicaltestsusedintheidentificationofBacilli:
Hemolysis
Tyrosinase:positiveontheleft,ontheright
negative
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Lesitinaasitesti,Thepositiveleft,right,anegativeresult.Thetestmeasures
munankeltuaisagarinsisltmnlecithindegradationofglycerolandfattyacids.Fattyacids
precipitatefromthereactionzone
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B.cereusATCC14579:TEMimage
negatiivivrjtyistspores.Ranad
Shaheen,2008.Bar=1microns
B.cereussporesofthebacteriumresistanttomany
heating.Particularlydurableare
cereulide(oksetustoksiini)producing
Sporesofstocks.
B.cereusF4810/72:TEMimagespores
thinsection(MariaAndersson)
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ScanningElectronMicroscope
(SEM)imagesB.cereusbacterium
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sporesadheredtothesteelplate
surface.Theleftimageshows
B.cereus(sensulato)isthesurfaceofthebacterialspores
steelsurfacegranularity(granulelimits).
hydrophobic,i.e.waterrepellent.Thereuponit
RanadShaheen,2008.
sticksoutofthewaterorfoodhydrophobic
surfaces,especiallyplasticsandsteel.
Adheredtothesurfaceofthesporeisdesinfioitavuus
poorbecausethesurfaceoftheadheredsporesareresistantto
heatandchemicaldisinfectionmore
thesporessuspendedintheliquidphase.
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CharacteristicoftheTablecereulideProducingB.cereussporestestedinpaperIIIandIV
Source:RanadShaheen,PhD,
UniversityofHelsinki,DissBiocentr.Molec.
UnivHelsigiensisinVik34/2009
Cereulide(B.cereus
Cereulide
Cereulide
Numberof
Producing
non
cereulide
B.cereus
ProducingB.
Producingstrains
cereus
OutofThosetested
370
90
17/81
25120
2530
17/81
inthattheirspores
17/81
resistantheating
Resistanttoheatat90C
D(min)value
(min)value
log10Viabilitylossafter120min
producedbythebacterium
oksetustoksiini)
producingstrains
differfromotherB.
cereusstrains
IDandanalkalinewash
at90C
multiply
Log 10 (N 0/N)germinationat7
Cafter7days
0.2
1.82
17/81
Log 10 (N 0/N)germinationat30
Cafter50min
1.8
1.82.5
17/81
morethan
"Ordinary"antidiarrheal
diseasescausedbyB.
cereusstrains.
Particularlydifficultis
Resistanttohotwashingwith:
theirhightolerance
hotalkalinewash
Alkaline(1%NaOH+75C,
pH13.1)
RangeD(min)initial
11.413.6
1.167
3/20
andnitricacid
1.42.5
0.15
3/20
flushing,whichare
aremm.dairies
Rangeoflogkill15min
usedequipment
Acid(0.9%,65C,HNO3)
RangeD(min)initial
3.3
1.325
1/6
Rangeoflogkill15min
3.5
0.45.9
1/6
Cleaningmethods
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Cfu/ml 10 9
Whenthefoodproductispasteurized(eg.Milk,
72C)suchasB.cereusspores,exceptthat
10 8
retaintheirviability,pikakynnistvt
germination,sothatthevegetativeB.cereus
theconcentrationofbacteriainthefoodrises
10 7
eg.10100times.Ifnot
heatedwould,butthefood
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bemaintainedatalowertemperature,
sporegermination(germination=)starts
10 6
onlyslowly:forexample,7C.inmaytake
aweekbeforeeven10%ofthosepresent
getssporestogerminate.FrozenPastry(eg.
10 5
C)in
Karelianpasties)omapaisto(250
initiatesporegermination,ietheproduct
containedinB.cereucanquicklygs
10 4
multiplyandcausefoodpoisoning,
unlessthepietoeatsoon.
However,pasteurizationisoftennecessary
10 3
otherfoodpollutersandhumanhealthhazards
causingthedestructionofbacteria(eg.
mycobacteria,listeria,salmonella).
10 2
0
1
2
3
Cookingtime,minutes
10
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Bacilluscereusproducesmanydifferenttypesofproteintoxin.Theyareproducedby
fisheatenfoodgastrointestinalbeenvegetativebacteria,
Sporesandtheirdescendants.Startingthisway,diarrhealdiseaseisinfectiousand,therefore,
Cereulidethatis,
diseases(suchassalmonellosis,campylobacteriosis,yersiniosis,etc.),but
oksetustoksiinia,
usuallyofshortduration.
producingB.cereus
Table4.PhenotypicandgenotypicpropertiestoreportedasspecificforthecereulideProducing
positionsdiverge
strainsofB.cereus.
ripulitoksisistainthat
theyproducethetoxin
Properties
Phenotypictraits
Reference
weakhaemolysis
Anderssonetal.,2004
inabilitytostarchhydrolysis
Shinagawaetal.,1979
salicindecompositionnegative
Shinagawaetal.,1993
H1predominanceofserovar
Agataetal.,1996
lackoftheHaemolyticenterotoxin
Agataetal.,1996
formedalreadyinourfoodand
causeeatenalmost
immediatepoisoning
(toksikoosin).Cereulide
producingstrains
differinanumber
physiologicaland
genetic
properties
Production
otherB.
Genotypictraitsspecificribopattern
specificRAPDprofiles
Pirttijrvietal.,1999
cereusstrains.They
EhlingSchulzetal.,2005a
aremm.usually
hemolysisare
specificSDSPAGEprofilesexoproteinEhlingSchulzetal.,2005a,
Identical16SrRNAgenesequences
IdenticalMLSTsequencetype
IdenticalFTIRspectra
todegradestarch,and
growhigher
attemperaturesotherthan
B.cereugs(upto+
Until52C)
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Starch
hydrolyysikyky
(amylolyyttisyys)is
generallypositive,aswellas
B.cereusgroupthatB.
subtilisgroup
bacteria.
salicin
Salicin(orthohydroxybenzylalcohol)isusedasthetestsubstancebetaglucanase
fortestingtheactivity.Betaglukanaasiwhereintheenzymehydrolyzesanetherbond
formedsalisyylialkoholia.
Cereulideproducingstrainsareusuallybothtrkkelysnegatiivisiathatsalisiininegatiivisia,
So,unlikeotherB.cereus.
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Bacilluscereusoksetustoksiini:cereulidemoleculeproperties.Cereulideinwater
insoluble,lipophilicmolecule,whichcarriespotassiumjonejathroughbiologicalmembranes,so
troublemakerse.g.humanbodypotassiumhomeostasis.Itisalsoamitochondrialpoison.Cereulide
withstandwithoutlosingitsactivityautoclaving,acidandalkalitreatment,drying.
Molecularstructure
Cyclicdodecadepsipeptide,1152g/mol
Synthesis
Nonribosomal
Sensoryproperties
ColourLess,odorless
Octanolwatercoefficient
LogKow6.0
Heatstability
Nolossofactivityuponcookingorautoclaving
pHstability
Stablebetween211
Blacklipidmembrane
Potassiumionophore
Isolatedratlivermitochondria
CatalysesafluxofK+ions
Modeofactioncell
Depolarizedmitochondrialmembrane
ofboarspermcells
ofNKcells
ofneuralcells
ofisolatedratlivermitochondria
Cereulidestructure1152g/molpeptiditoksiini:
cyclo(OValLValODLeuDAla)3
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Exposedcereulidepig
spermJC1colored.
Ispartofthemitochondria
lost
andtheelectricchargeofthesperm
hyperpolarisoitunutend.
Spermmitochondriaarelocatedsiittihnnn(=flagellin),thebasepart.Ofthespermheaddoesnothaveany
mitochonidria.Thisuniquearchitectureallowsthecellmitochondrialdamageeasy
Detectionbylightmicroscopy,e.g.,JC1stainedcellswithafluorogenic.FigureswimJC1
colored,healthypigspermatozoawithhead(=lowmembranepotential)fluorescesgreenandthetail
thebaseofthemitochondriaorange.
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BacillussubtilisbacterialspeciesgrouplivinginthesameenvironmentsasB.cereus.
Table7.EnvironmentalnichestoreportedforthemembersofB.subtilisgroup
Species
soil a
water b
food c
mammalian
gut d
+
insect
gut e
+
Industrial
fermentations f
+
+
B.subtilisssp.subtilis
+
+
+
B.amyloliquefaciens,
+
B.atrophaeus
+
B.licheniformis
+
+
+
+
+
B.mojavensis
+
+
B.pumilus
+
+
+
B.subtilisssp.spizizenii
+
B.vallismortis
+
+
B.sonorensis
+
+
a PriestF.1989Nakamuraetal.,1999Roberts,etal.,1996aNakamura1989Roberts,etal.,1994
intherhizosphereor
as
endophyteofplants g
+
+
+
+
+
+
Palmisanoetal.,2001
b PriestF.1989stensviketal,2004..Jeongetal,2007
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c PriestF.1989Frometal,2005..Sorokulovaetal,2003
d Tametal.,2006Leseryou.al.,2007
eKnigH.2006
f Ochiaietal.,2007PriestF.1989
g Idrissetal.,2002BaconandHinton,2001Manoetal.,2006Parketal.,2007
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Bacillusamyloliquefaciensoffoodbornebacterialtoxin
Amylose.
Amyloseisahydrophobicpeptide,1197Da,composedofleucine,proline,
serine,asparticacid,glutamicacidandtyrosine.Itisidentified_byandquantitated
basedonitsmolecularions(1197m/z)
AmyloseFormedcationselectivechannelsinlipidmembranes,witha
selectivityK
+ Na
+ Ca
2+ of26:15:3.5The
ExposuretoAmylosedepolarizedthetransmembranepotentialsofmitochondria
(
m)andoftheplasmamembrane(
p)ofprimaryboarsperm,felinelung,human
NKcellsandcelllines(lungCalu3,neuralPaju)uponwithin20minofexposure
Amyloseproducespotassium,sodiumandcalciumjonejapermeabletroublemakerchannels
mammaliancells.Innateimmunecells(macrophages)Amylose
sytokiinimyrskynstart,ifpresent,isatthesametimesomeofthemicrobialcell
componentssuchase.g.lipopolysaccharide(LPS).
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Bacillussubtilisheatstabletoxinsproducedbythegroup:
Testcellsperm(swine)
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Humantestcell
colon
Adenocarcinoma
(Caco2)
Page19
IonophoricactivityofAmylosefromBacillusamyloliquefaciens
measuredbyblacklipidmembranes(BLM)
alogofelectricconductivity
106
Permeabilityratio
ofchannelsfor
cationsover
anions:
K + /Cl
ofthelipidmembrane
105
>26
2104
m
c
/
S103
n
,
ec
n102
as
c
u101
d
n
No
C100
logconcnofsubstance
Na
+ /Cl
Ca
2+
/Cl
=15
=3.5
ionchannels
101
0.1
1
Concentration,micrg/ml
30
Voltageacross
25
themembrane
KCI
NaCl
20
V 15
m
10
CaCI
5
0
Tefloncuvettewith50um
membranecoveredwindow
0.0
0.2
0.4
0.6
concentrationratioofions
thelogoftheconcentrationratio
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JC1stainingRevealstheeffectsoftheHPLCpurified
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Amyloseonboarspermcellsandhumancolon
epithelialcelllineCaco2
Boarsperm
Caco2
Amylosedepolarisedthemitochondrialand
plasmamembraneelectricpotentials:
StructureofAmylosewasRevealedby
MS/MSandH1NMR
Page21
Amyloseisadangersignaltothehuman
macrophages
Dangersignal
LPS
Source:StiinaRasimus&
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SampsaMatikainen
Page22
TheEUandEFSA(European
FoodSafety
Agency)compiledby
amassivedatabase
foodpoisonings
Europeshows
theBacilli,
inparticularBacillus
cereup,theroleis
Europe,significant
toxic
foodpoisoning
acause.
Toxicfoodpoisoningmeansthataman
makingsicktoxinwasalreadypresentinthefood.
Livemicrobesisnotnecessarilyfoundinfood(can
bekilledbyheating).Toxic
ruokamyrkytykstenmostcommoncausesare
fromtheEU/EFSAreport(theks.oikeallalist).
Otherfoodpoisoningsaremainlyinfections,ie,
microbial(eg.Bacilluscereus)hasenteredthefoodor
beveragewithgastrointestinalbegunand
theremultiplyandproducetoxinsor
virulencefactorsofwhichisthehumanbody
disadvantage.
Page23
Bacilli
and
standards
Bacilluscereus
determination
foodis
ThetwoISO
Standard:SFS
ENISO7932
(nextdoor)and
ISOEN21871:
Microbiologyoffood
andanimalfeeding
stuffshorixzontal
Methodforthe
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determinationof
presumptiveBacillus
Thus,foodshavenotbeenthresholdsB.cereusconcentration.Specialgroups
cereus.Most
(infants,immunocompromised)thepreparationoffood
ProbableNumber
processhygienemustbeofsuchqualitythattheB.cereusspores
(MPN)technique
contentdoesnotexceed50m=(satisfactory),M=500(acceptable)cfu/g.(EU
anddetection
CommissionRegulationNo1441/2007.Off.J.Eur.Lunion,322:1229).
method.Colony
countingtechnique
at30C.2005
Page24
Bacillussporesaredifficulttodisinfect.Sporesbitesglutaricandopalaldehydes.Theeffectdependson
pHandcontacttimewillbelong.Duetotoxicresiduesofthesesubstancesarenotsuitable
ofthefoodprocessingenvironment.
Bacillussubtilisspores
60
120
Escherichiacoli
180min
10
pH4
1
0.1
pH7.9
0.01
Glutaaridialdehydi,0.0044%
Glutaaridialdehydi2%
GTAalsoinactivatesvirusesandspores.Dangerousbyinhalation,allergenic,tosensitize.
OPAismoreeffectiveformycobacteriaandfatsolublesubstancesinanenvironmentlikeGTA,
apopulardisinfectantinhospitals,especiallyequipmentmaintenance,aneffectivemicrobicideandsporisidiand
moreresistanttoorganicdirtloadthanmanyothermaterials.Itdoesnotattackrubber,metals,
plastics,instrumentsorotherinstruments.
GTAishighlytoxictohumans,skincontactandinhalation.
OPAisaseffectivemicrobicidethantheGTA,butweakersporisidi.Itissafertotheuser
asGTA(evaporatesless).
Page25
DisinfectantsBacillusspores.
Chart2.2.4b.Sporeproducingpowerofperaceticacid
bacteria.Thefiguresrepresenttheminimumkilltime
7 cfuml
nuutteina.Bacterialinitialconsistencyof10
Bosher1987,andPaulus1993basedonthefacts.
Chart2.2.4.aBacillussubtilissporesPRINCI
BacterialSpecies
hydrogenperoxidetolerance.
Ksittelyl
mptila
AccordingtoW.Paulus(1993)
H
Pit%
Survival%,and
Bacilluscereus
exposuretime(min)
2
30
60
Peraceticacid
contentinmgl
200
>60
20
>60
40
40
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85
22
2.1
10
35
0.0027
15
22
0,0022
Bacillussubtilis
Clostridium
perfringens
5
20
>60
10
40
10
>60
20
>60
40
2.5
CereuPRINCIBacillussporesaremoredurablethanBacillussubtilis.Foodand
thepackagingindustryusedforthedisinfectionofsporesofhydrogenperoxide(bestpowerwill
heated)andperaceticacid,whichissoldinaceticacid,peraceticacidand
hydrogenperoxidetasapainoseoksena.Ofthese,donotleaveanytoxicresiduestotheenvironment.
CereulideproducingstrainsofB.cereussporesstocksareevenmoredurablethanotherB.
cereusten.
Page26
StandardmethodsforsporisidisenpowermeasurementI.
Thesporesfordisinfection
soldmaterial
sporisidinen(=spores
Lethal)power
measurable
standardized
methods.
Elintarvikeymprists
Theweatheruse
theintendedsubstances
powerstudy
appended
standards,
oftestis
accordingtothestandard
B.usesubtilis
DSM347(=ATCC6633)
orB.cereus
CIP105151(=ATCC
12826)spores.
Surfacedisinfection
protocolsfor
sporisideilleis
separatemethods.
Page27
Standardmethodsforsporisidisenpowermeasurement2.
15/16
9/13/2015
Bacilliinfood
Sporisidinenpowerbyanumericalvaluelog[N
/N],whereN
incontrol(=lumeksitelty,orusedasdisinfectantneutralizinginactivated)andN
istheamountofsporeswithgrowth
istheamountofsanitizingprocessingpassesthroughthesample.OutputNumber=0means,therefore,
thesituationinwhichbothhadthesamenumberofviablesporeinactivationisnotatapahtnunut.
Sporisidinenpowerofthechemicalishighlydependentonthedurationoftheprocessingoperation,temperature,pH
,theactivesubstanceconcentration,andinthedegreeofcontamination(dirtconsumingdisinfectionefficacy).
Researchingthepowerofsubstancesintendedforhumanandanimalskindisinfectionisnotsafety
reasonsusedBacillussporesexperimentaleliin.AccordingtoEN1499:1997,isusedtoE.coliK12
position(=tamedversionofthehumangutbacteriumE.coli"to).Itdoesnotproducespores.Themostcommonlyused
skindisinfectant,70%ethanol,spikedwithorwithoutalowconcentrationofchlorhexidine.Skin
isinvolvedintheprotectionofglycerol.Sporisidistakinthistreatmentiseffectiveifandwhentheskinisnot
aftertreatment,rinsewithwater,butlettheethanolevaporate.
16/16

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