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nocelmultiplic剖ionofstrainATCC33152wasobserved was48.40CwhenthebacteriawasgrowninBCYE-αbroth withoutcharcoalfor6Qh.Thisdiferenceincriticaltemper目 aturewasprobablydueto:(i)ag訂 beingusedratherthan bro出， (i)moreacuratetempera加recontrolyviththeTGI apara加 susedinourexperiment.Itcouldbeextrapolated thatwhenthewatertempera加 reinarecirculatingwatersup- plysystemiskepthigherthanat4.1oC，morethan88.9% ofwilds廿ainswouldnotsurvive，andthatwhenthetemper- a旬reiskepthigherthanat4.50C，nostrainswouldsurvive. However，thisex仕apolationwouldnotaplytobacteriaem- beddedinabiofilmstructure，because.thebiofilmwould protectthebacteriafromunfavorablegrowthconditions， andamuchhighertemperaturewouldbeneededtoki1lany bacteriaembeddedinthebiofilm.Inthiswork，bacteria weregrowninarichmedium(BCYE-α)whereasthereal- wor1dsystemofahotwaterdistributionsystemisexpected tobeoligotrophic，inwhichthenutrientconcentrationsare perhapstwoordersofmagnitudesmaler.Inthenextfu旬 re work，wewouldliketoverifシtheheterogeneousstructureof Legionellabiofilmbyusingbiofilmflowcelsystem，in whicholigotrophicenvironmentcanbedemonstrated.
ACKNOWLEDGMENTS
Thisresearchwassponsoredinpartbyagrant-in-aid(nos. 15037and1750324)forScienceResearch企omtheMinistry ofEducation，Cul同re，Sports，ScienceandTechnologyofJapan.
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