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Incubation Numberof
temp.(OC) colonies Average S.D Average S.D. Average S.D.
37 103 39 56 27 15 37 15 40 58 189 77 99 37 52 12 42 15 359 85 154 21 74 8
50 0 r l -
sE 400トl {〉O口B、301ト
Y:
Z3 2 0 0 匂04
t10巳戸
4 0 E2 /~ 斗20
iω 1∞
O~
.
T
百
，- 10 0 →80
v 」。 36 38 40 42 44
Incubationtemperaturetc) Fig.7EfectsofincubationtemperatureonL.pneumophilabiofilmproperties.
SymbolsofPvaluefort-testingare牢:P<0.05and本* :P<0.0l.
Fig.8AshowsthelightmicroscopyphotographyofabiofilmofL.pneumophilain liquidflowofBCYE-αmediumat370C.Thebiofilmcomposedofmicrobialasemblages ofL.pneumophilawasobserved.Fig.8Bshowsmagnifiedlightmicroscopicphotographyof Fig.8A.Thebiofilmexhibitedatypicalholowmicrocolonywhichwascomposedofshort rodbacteria.Fig.8CshowsthephotographofthickL.pneumophilabiofilmdevelopedon theglaswal，ontheair-liquidat400C(16). Fig.8Dislightmicroscopicphotographyof Fig.8C.Thebiofilmexhibitedatypicalfilamentousmorphologythatdidnotdifermuch whenever the bacteria were located in the biofilm.Fig.8E shows the streak ofL.
pneumophiladevelopedonBCYE-αagarplateat370C.Thelightmicroscopicexhibiteda typical filamentous morphology in Fig.8F. Table 3 is a comparison of L. pneumophila bacterialengthofabiofilminliquidflowwithBCYE-αmedium，ontheglaswalinstatic liquidandonBCYE-αagarplate.Theaveragelengthwas4.4土l.7μmat370C，2.9土l.2 μmat40oC，3.5土 2.0μmat420Cintheliquidflow，respectively.Theaveragelengthwas
28士 15μmat370C，50士20μmat400Cand80士26μmat420Cinstaticliquid
respectively.Theaveragelengthonagarplatewas58士 36μmat370C，98土 44μmat400C and125土 68μmat420C，respectively.Figure9showsthelengthofL.pneumophila bacteria in static liquid and on BCYE-αagar plate increased with the temperature， respectively.However，thelengthofL.pneumophilabacteriainliquidflowdidnotdifer withthetemperature.
4.Discussion
MicrocoloniesofL.pneumophilabiofilmsonglassurfaceinliquidflowchanelwere observed by confocal laser scaning microscopy (Fig.4A). The average number of legionelabacteriapersinglecolonyincludes4.9x104inP2，l.3x105inP3and1.2x104
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