施設内感染を起こす緑膿菌の水系環境における生息実態と病原性保有の

Შ଀ᚨϋज़௨ǛឪƜƢዯᐝᓏƷ൦ኒ࿢‫ؾ‬ƴƓƚǔ
ဃऒܱ७Ʊ၏Ҿࣱ̬ஊƷᚐௌ
٤྾ችඳੱ㧔ᗲᇫ⋵┙ⴡ↢ⅣႺ⎇ⓥᚲ㧕
‫⎇ޣ‬ⓥ⋡⊛‫ޤ‬
ᣣᧄߥߤߩක≮వㅴ࿖ߢߪ㧘ᒝᲥ⩶ߦࠃࠆ޿ࠊࠁࠆવᨴ∛ߪᷫዋߒߚ߇㧘ᒙᲥ⩶ߦࠃࠆ
ᣣ๺⷗ᗵᨴ∝߿⮎೷⠴ᕈ⩶ߦࠃࠆᗵᨴ∝ߪ߻ߒࠈჇടߒߡ޿ࠆ‫ߪ⩶⤥✛ޕ‬ᄙߊߩᶖᲥ೷߿
᛫⩶೷ߦ⥄ὼᛶ᛫ᕈ߇޽ࠆ਄㧘ᓟᄤ⊛ߥ⮎೷⠴ᕈ߽₪ᓧߒ߿ߔ޿ߚ߼㧘޿ߞߚࠎ⊒∝ߔࠆ
ߣᴦ≮߇࿎㔍ߥ႐ว߇ᄙߊ㧘ᗵᨴ㒐ᓮ⢻ߩૐਅߔࠆ⠧ੱ߿♧ዩ∛ߥߤߩၮ␆∔ᖚࠍ᦭ߔࠆ
ੱߦߪ⢿ᆭߢ‫߽ᦨޔ‬㊀ⷐߥ∛㒮╬ߩක≮ᣉ⸳ౝᗵᨴේ࿃⩶ߩ㧝ߟߢ޽ࠆ 1)‫ߦ․ޕ‬㧘ᢌⴊ∝ࠍ
⊒∝ߒߚ႐วߩ⥌๮₸ߪ 80%એ਄ߢ㧘⥃ᐥ਄㊀ᄢߥ໧㗴ߣߥߞߡ޿ࠆ‫ޕ‬
✛⤥⩶ߪ‫↢ޔ‬ᵴ࿤޽ࠆ޿ߪ⥄ὼ⇇ߩ᳓♽ⅣႺߦᐢߊሽ࿷ߔࠆⅣႺᏱ࿷⩶ߢ㧘ක≮ᣉ⸳ౝ
ߦ㒢ቯߒߚኻ╷ߩߺߢߪ㒢⇇߇޽ࠆ‫ޕ‬᳓♽ⅣႺ߇ਥߚࠆᗵᨴḮߢ޽ࠆߦ߽߆߆ࠊࠄߕ‫✛ޔ‬
⤥⩶ߩ↢ᘒߦߪਇ᣿ߥὐ߇ᄙ޿‫੹ޕ‬࿁‫∛ޔ‬㒮ᖚ⠪߅ࠃ߮๟ㄝߩ᳓♽ⅣႺ߆ࠄಽ㔌ߐࠇߚ⩶
ᩣࠍ↪޿ߡ‫ޔ‬ⅣႺਛߢߩ↢ᕷታᘒ‫ޔ‬ㆮવሶဳ‫∛ޔ‬ේㆮવሶߩ଻᦭⁁ᴫߥߤࠍ⺞ߴߡ‫ޔ‬᳓♽
ⅣႺߩẜ࿷࡝ࠬࠢࠍᛠីߒ‫ޔ‬࿾ၞߩ଻ஜⴡ↢࡮ᗵᨴኻ╷ߦ⾗ߔࠆߎߣࠍ⋡⊛ߣߔࠆ‫ޕ‬
‫᧚ޣ‬ᢱߣᣇᴺ‫ ޤ‬
㧝㧚᳓♽ⅣႺ߆ࠄߩ✛⤥⩶ಽ㔌
ᗲᇫ⋵ߩᴡᎹ㧘ᴧḓ㧘ᴪጯߥߤ߆ࠄ᳓ࠍណขᓟ㧘⚦⩶↪ၭ࿾ߢ↢⩶ᢙࠍ᷹ቯߒߚ‫⤥✛ޕ‬
⩶ߩㆬᛯߪ㧘࠮࠻࡝ࡒ࠼ၭ࿾ߦ࠽࡝ࠫࠢࠬ㉄ࠍᷝടߒߚ߽ߩࠍ↪޿ߡⴕߞߚ‫ߩ⩶⤥✛ޕ‬ห
ቯߪ㧘↢‛ቇ⊛࡮↢ൻቇ⊛ᕈ⁁⹜㛎ࠍቯᴺߦߒߚ߇ߞߡታᣉߒ⏕⹺ߒߚ‫⊖ޕ‬ᢙචᩣߩ✛⤥
⩶ᩣࠍಽ㔌ߒߚ‫∛ޕ‬㒮ᖚ⠪߆ࠄណขߐࠇߚ⥃ᐥಽ㔌⩶ࠍኻᾖߣߒߡ↪޿ߚ‫ޕ‬
㧞㧚✛⤥⩶ᩣߩㆮવሶ࠲ࠗࡇࡦࠣ
ㆮવሶဳ࠲ࠗࡇࡦࠣߪ㧘ߘࠇߙࠇߩ⩶ᩣࠃࠅࠥࡁࡓ DNA ࠍ᛽಴ߒ㧘೙㒢㉂⚛ߢᶖൻߒߚ
ᓟ㧘ࡄ࡞ࠬࡈࠖ࡯࡞࠼㔚᳇ᵒേࠍታᣉߒߚ‫↹ࠍࡦ࡯࠲ࡄ࠼ࡦࡃߩߘޕ‬௝⸃ᨆߒߡ㧘⋧หᕈ
ࠍၮߦ ࠣ࡞࡯ࡇࡦࠣࠍⴕߞߚ‫ޕ‬
㧟㧚✛⤥⩶ᩣߩ∛ේㆮવሶ଻᦭⁁ᴫ
✛⤥⩶ߩ∛ේᕈ⊒⃻ߣ㑐ㅪߔࠆࠢࠝ࡜ࡓ࠮ࡦࠪࡦࠣ(Quorum Sensing: QS)ㆮવሶ(lasI,
lasR, rhlI, rhlR)߅ࠃ߮࠲ࠗࡊ III ಽᴲᯏ᭴(Type III Secretion System: TTSS)ㆮવሶ(popB,
㧙㧙
popD, pcrV, exoS, exoT, exoU, exoY)ߩ଻᦭⁁ᴫࠍ‫ޔ‬ฦ⩶ᩣߩࠥࡁࡓ DNA ࠍ㍌ဳߣߒߡ
PCR ᴺߢ⺞ߴߚ‫ޕ‬
‫⚿ޣ‬ᨐߣ⠨ኤ‫ޤ‬
㧝㧚⥃ᐥ࡮ⅣႺಽ㔌ᩣߩㆮવሶဳߩ․ᓽ
ᗲᇫ⋵ౝߩᴪጯ࡮ᴡᎹߥߤߩ⥄ὼⅣႺ߅ࠃ߮∛㒮ᖚ⠪߆ࠄ✛⤥⩶ࠍಽ㔌ߒߚ‫ޕ‬ណขߒߚ
႐ᚲ㧘߅ࠃ߮ಽ㔌ߒߚ✛⤥⩶ᩣࠍ࿑㧝ߦ␜ߔ‫⩶ޕ‬ᩣ㑆ߩࠥࡁࡓㄭ✼ᐲࠍ⍮ࠆߚ߼ߦ㧘ฦ⩶
ᩣ߆ࠄᨴ⦡૕ DNA ࠍ᛽಴ߒ㧘೙㒢㉂⚛ SpeI ߢಣℂᓟ㧘ࡄ࡞ࠬࡈࠖ࡯࡞࠼㔚᳇ᵒേ(PFGE)
ࠍታᣉߒߚ‫ࠄ߆ࡦ࡯࠲ࡄ࠼ࡦࡃߩߘޕ‬㧘ࠥࡁࡓႮၮ㈩೉ߩㄭ✼ᐲࠍ⸃ᨆߒߚ㧔࿑㧞㧕‫ޕ‬60%
⋧หᕈࠍၮߦ 13 ࠣ࡞࡯ࡊߦ߹ߣ߼ࠄࠇߚ‫⥃ޕ‬ᐥᩣߪోߡ߇⇣ߥࠆ PFGE ࡃࡦ࠼ࡄ࠲࡯ࡦࠍ
␜ߒㆮવሶဳ߇ᄙ᭽ߢ޽ߞߚ‫ߪߣߎߩߎޕ‬㧘ห৻∛㒮ౝߢಽ㔌ߐࠇߚ⩶ᩣߢ޽ࠆ߇㧘㒮ౝ
ᗵᨴߦࠃࠆવ᠞ߢߪߥߊ㧘ߘࠇߙࠇߩᖚ⠪ߪ୘೎ߦᗵᨴߒߚߎߣࠍᗧ๧ߒߡ޿ࠆ‫ޕ‬ⅣႺᩣ
ߢߪ⥝๧޽ࠆ⍮⷗߇ᓧࠄࠇߚ‫ߜࠊߥߔޕ‬㧘ᴡᎹਛᵹၞ㧘ਅᵹၞߩಽ㔌ᩣߪ㧘ᄙ᭽ߥㆮવሶ
ဳࠍ␜ߒߚ߇㧘ᴡᎹ਄ᵹၞ㧘ᴡญၞ㧘߅ࠃ߮ᴪጯᶏ᳓ၞߩಽ㔌ᩣߪᄙ᭽ᕈ߇ૐߊ㧘ห৻ߩ
ࠢࡠ࡯ࡦߣᕁࠊࠇࠆ⩶ᩣ߽ሽ࿷ߒߚ㧔࿑ 2㧕‫ޕ‬ᴡᎹਛᵹၞ㧘ਅᵹၞߪಽ㔌⩶ᢙ߽ᄙߊ㧘ᄙ᭽
ߥ✛⤥⩶߇ᷙ࿷ߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ‫৻ޕ‬ᣇ㧘ᴡᎹ਄ᵹၞ㧘ᴡญၞ߅ࠃ߮ᴪጯᶏ᳓ၞߢߪ
㒢ࠄࠇߚㆮવሶဳߩ✛⤥⩶߇↢ᕷߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ‫ߚ߹ޕ‬㧘ਅᵹၞ㧘ᴡญၞ㧘ᴪጯၞ
ߩಽ㔌ᩣߩㆮવሶဳ߇⇣ߥߞߡ޿ࠆߎߣߪ㧘ᵹࠇߦߒߚ߇ߞߡᵹ౉ߒߚ⩶߇ߘߩ߹߹ቯ⌕
ߒ↢ᕷߔࠆ⸶ߢߪߥߊ㧘ᴡญߩᴁ᳓ၞ߿ᴪጯߩᶏ᳓ၞߦㆡᔕߒߚ߽ߩ߇↢ᕷߒߡ޿ࠆߣ⠨
߃ࠄࠇࠆ‫ޕ‬
㧞㧚⥃ᐥ࡮ⅣႺಽ㔌ᩣߩ∛ේᕈ㑐ㅪㆮવሶ଻᦭⁁ᴫ
✛⤥⩶ߩ∛ේᕈߦ㑐ߔࠆ⎇ⓥߪ㧘ਥߣߒߡ⥃ᐥಽ㔌ᩣࠍኻ⽎ߦታᣉߐࠇߡ߈ߚ‫ࠄࠇߎޕ‬
ߪࡅ࠻ߦᗵᨴߒჇ⩶ߢ߈ߚ⩶ߢ㧘∛ේᕈ߇㜞޿߽ߩ߇ㆬᛯߐࠇߡ޿ࠆน⢻ᕈ߇޽ࠆ‫⥄ޕ‬ὼ
ⅣႺਛߦ↢ᕷߔࠆ✛⤥⩶ߩ∛ේᕈߪߤ߁ߢ޽ࠈ߁߆‫ߩߎޕ‬ὐࠍ⺞ߴࠆ⋡⊛ߢ∛ේㆮવሶߩ
ሽ࿷ߩ᦭ήࠍᬌ⸛ߒߚ‫∛ޕ‬ේㆮવሶ୥⵬ߩਛ߆ࠄ㧘ࠢࠝ࡜ࡓ࠮ࡦࠪࡦࠣ(QS:Quorum
sensing)ㆮવሶ(lasI, lasR, rhlI, rhlR)㧘߅ࠃ߮࠲ࠗࡊ III ಽᴲᯏ᭴(TTSS:type III secretion
system)ㆮવሶ(effector ㆮવሶ: exoS, exoT, exoU, exoY, ߅ࠃ߮ translocator ㆮવሶ: popB,
popD, pcrV)ߩ 11 ୘ߩㆮવሶࠍኻ⽎ߣߒߚ‫∛ߩ⩶⤥✛߇ࠄࠇߎޕ‬ේᕈ⊒⃻ߦ߅޿ߡ㊀ⷐߢ
޽ࠆߎߣ߇㧘⥃ᐥᩣߩ⎇ⓥ߆ࠄ᣿ࠄ߆ߦߐࠇߡ޿ࠆ‫ޕ‬
QS ᯏ᭴ߪ⚦⩶߇๟࿐ߩ⩶ኒᐲࠍᗵ⍮ߔࠆᯏ᭴ߢ㧘⩶ኒᐲ߇৻ቯએ਄ߦߥࠆߣ․ቯߩㆮવ
ሶߩ⊒⃻߇㐿ᆎߐࠇࠆ 2)‫ޕ‬QS ᯏ᭴ߪ⩶ߩ∛ේᕈ⊒⃻ߦ߅޿ߡ㊀ⷐߢ㧘ᗵᨴዪᚲߢ⩶߇৻ቯ
ᢙჇᱺߒߚᲑ㓏ߢ∛ේ࿃ሶߩวᚑ߇㐿ᆎߔࠆߎߣߪ㧘∛ේ૕ߦߣߞߡวℂ⊛ߣ⠨߃ࠄࠇߡ
㧙㧙
޿ࠆ 3, 4)‫৻ޕ‬ᣇ㧘ᴡᎹ߿ᴪጯߩ᳓ၞߪੱ૕ߥߤߣᲧߴߪࠆ߆ߦ⽺ᩕ㙃⁁ᘒߢ޽ࠅ㧘ណขߐࠇ
ࠆ⩶ᢙ߽ዋߥߊ㧘⩶ኒᐲ߇ߐ߶ߤ㜞ߊߥࠆߣߪ⠨߃ߦߊ޿‫ߡߞ߇ߚߒޕ‬㧘⥄ὼⅣႺࠍਥߚ
ࠆ↢ᕷ႐ᚲߣߔࠆ⩶ߪ㧘QS ㆮવሶߩᔅⷐᕈ߇ૐ޿ߩߢߪߥ޿߆ߣ੍ᗐߐࠇߚ‫ߒ߆ߒޕ‬㧘Ⅳ
Ⴚᩣߪ⥃ᐥᩣߣห᭽ߦ QS ㆮવሶࠍ଻᦭ߒ㧘⺞ߴߚ 154 ᩣߪోߡ QS ㆮવሶ(lasI, lasR, rhlI,
rhlR)ࠍ᦭ߒߡ޿ߚ㧔⴫ 1㧕‫ߪߣߎߩߎޕ‬㧘QS ᯏ᭴߇∛ේᕈ⊒⃻ߦ㑐ਈߔࠆߩߺߥࠄߕ㧘⥄
ὼⅣႺਛߢߩㆡᔕ࡮↢ሽߦ߅޿ߡ߽㊀ⷐߥᓎഀࠍᨐߚߒߡ޿ࠆߎߣࠍ␜ໂߔࠆ‫ޕ‬
TTSS ߪ࠾࡯࠼࡞᭴ㅧࠍㅢߓߡᮡ⊛ኋਥ⚦⢩ߦࠛࡈࠚࠢ࠲࡯࠲ࡦࡄࠢ⾰ࠍᵈ౉ߔࠆᯏ᭴
ߢ㧘ࠛࡈࠚࠢ࠲࡯ߪ⚦⢩↢ℂᯏ⢻ࠍᡬੂߒ㧘∛ේᕈߩ⊒⃻ߦ߅޿ߡ㊀ⷐߢ޽ࠆ 5)‫ޕ‬TTSS ߪ
⚦⩶ߩჇᱺߦߪᔅ㗇ߢߥߊ㧘ኋਥߦᗵᨴߒߚ㓙ߦᮡ⊛⚦⢩ߦ௛߈߆ߌࠆߩߢ޽ࠆ߆ࠄ㧘ᴡ
Ꮉ࡮ᴪጯߩ⥄ὼⅣႺਛߢߪᔅⷐᕈ߇ૐ޿ߩߢߪߥ޿߆ߣ੍ᗐߐࠇߚ‫ߒ߆ߒޕ‬㧘154 ᩣߩⅣႺ
ᩣ߅ࠃ߮⥃ᐥᩣߪోߡ TTSS translocator ㆮવሶ(popB, popD, pcrV)ࠍ଻᦭ߒߡ޿ߚ㧔⴫ 2㧕‫ޕ‬
effector ㆮવሶߦ㑐ߒߡ߽㧘ోߡߩᩣ߇ exoT ࠍ଻᦭ߒ㧘߶ߣࠎߤߩᩣ߇ exoY ࠍ଻᦭ߒߡ
޿ߚ㧔⴫ 2㧕‫ߡߞ߇ߚߒޕ‬㧘TTSS ߽⥄ὼⅣႺਛߢᯏ⢻ߒߡ޿ࠆߣᗐቯߐࠇࠆ‫৻ޕ‬ᣇ㧘exoS
ߣ exoU ߪ 2 ⠪ᛯ৻⊛ߦ଻᦭ߐࠇࠆ߇㧘ߘࠇߙࠇߩ଻᦭₸߇ಽ㔌႐ᚲߦࠃߞߡ⪺ߒߊ⇣ߥߞ
ߡ޿ߚ㧔⴫ 2㧕‫ߜࠊߥߔޕ‬㧘⥃ᐥᩣߢߪ exoS(+)exoU(-)߇ 64.5%,exoS(-)exoU(+)߇ 35.5%ߢ
޽ ࠆ ߩ ߦ ኻ ߒ 㧘 ᴡ Ꮉ ᩣ ߢ ߪ exoS(+)exoU(+) ߇ 1.1%, exoS(+)exoU(-) ߇ 98.9% 㧘
exoS(-)exoU(+)߇ 0%ߢ޽ߞߚ‫ޕ‬ㅒߦ㧘ᴪጯᩣߢߪ exoS(+)exoU(-)߇ 11.4%, exoS(-)exoU(+)
߇ 88.6%ߢ޽ߞߚ‫ޕ‬exoS ߪ N ᧃ┵ߦ GAP(GTPase activating protein)㗔ၞࠍ㧘C ᧃ┵ߦ
ADP ribosylase 㗔ၞࠍ᦭ߔࠆੑᯏ⢻࠲ࡦࡄࠢ⾰ߢ㧘ᮡ⊛⚦⢩ߩ↢ℂᯏ⢻ࠍᄌൻߐߖࠆ‫৻ޕ‬
ᣇ㧘exoU ߪ phospholipase A2 ᵴᕈࠍ᦭ߒ㧘ᮡ⊛⚦⢩ࠍ⚦⢩ᱫߐߖࠆ‫ޕ‬
‫ ޤߦࠅࠊ߅ޣ‬
✛⤥⩶ߩਥߚࠆ↢ᕷ႐ᚲߪ㧘⥄ὼ⇇ߩḨẢߥⅣႺߢ޽ࠆ‫ᦨߡߒߣ⩶⚦ޕ‬ᄢࠢ࡜ࠬߩࠥࡁ
ࡓ߿ᄙᢙߩ೙ᓮ♽ㆮવሶ࡮ឃ಴♽ㆮવሶߩሽ࿷ߪ㧘ⅣႺㆡᔕ⢻ജߩ㜞ߐߣ㑐ㅪ߇޽ࠆߢ޽
ࠈ߁
11)‫ߩߤߥ࠻ࡅޕ‬ኋਥߦኻߔࠆ∛ේᕈߩ⊒⃻ߦ․ൻߒߡ⊒㆐ߒߚߣ⠨߃ࠄࠇߡ߈ߚ∛ේ
ㆮવሶ⟲߇㧘⥄ὼ↢ᘒ♽ߢߩㆡᔕ࡮↢ሽߩㆊ⒟ߢ₪ᓧߐࠇߚߩߢߪߥ޿߆ߣ߽⠨߃ࠄࠇࠆ‫ޕ‬
߹ߚ㧘ߎߩࠃ߁ߥ⎇ⓥߪ㧘ߚߛߜߦ㧘ⅣႺᓸ↢‛ߩẜ࿷࡝ࠬࠢࠍ⹏ଔߔࠆߎߣߦߥࠅ㧘Ⅳ
Ⴚ߇ᗵᨴḮ࡮ᗵᨴ⚻〝ߣߥࠆࠃ߁ߥࡅ࠻ᗵᨴ∝ߩ੍㒐ᚻᲑߩ⏕┙ߦ߅޿ߡ߽㊀ⷐߢ޽ࠆ‫ޕ‬
‫ޣ‬ਥߥᢥ₂‫ޤ‬
1) Stover CK, Pham XQ, Erwin AL et al: Complete genome sequence of Pseudomonas
aeruginosa PAO1, an opportunistic pathogen. Nature 2000;406:959–64.
2) Bassler BL: Small talk. Cell-to-cell communication in bacteria. Cell 2002;109:421–4.
㧙㧙
3) Miller MB, Skorupski K, Lenz DH et al: Parallel quorum sensing systems converge to
regulate virulence in Vibrio cholerae. Cell 2002;110:303–14.
4) Wu L, Estrada O, Zaborina O: Recognition of host immune activation by
Pseudomonas aeruginosa. Science 2005;309:774–7.
5) Galán JE, Collmer A: Type III secretion machines: bacterial devices for protein
delivery into host cells. Science 1999;284:1322–8.
‫⻢ޣ‬ㄉ‫ޤ‬
ᧄ⎇ⓥࠍታᣉߔࠆߦ޽ߚࠅ㧘⎇ⓥഥᚑࠍ޿ߚߛ߈߹ߒߚ⽷࿅ᴺੱᄢห↢๮੐ᬺ࿅ߦෘߊ
ᓮ␞↳ߒ਄ߍ߹ߔ‫ޕ‬
‫૶⾌⚻ޣ‬ㅜౝ⸶‫ޤ‬
૶ ㅜ ౝ ኈ
⚦⩶ၭ㙃⹜⮎㧔᥉ㅢၭ࿾㧘࠮࠻࡝ࡒ࠼ၭ࿾㧘࠽࡝ࠫࠠࠪࡦ㉄㧕
ಽሶ↢‛ቇ⹜⮎㧔೙㒢㉂⚛㧘PCR ㉂⚛㧘วᚑࡊ࡜ࠗࡑ࡯㧕
ࡊ࡜ࠬ࠴࠶ౕེࠢ㘃㧔ࠪࡖ࡯࡟㧘㆙ᔃ࠴ࡘ࡯ࡉ㧘ࡇࡍ࠶࠻ࡑࡦ㧕
ߘߩઁ㧔ࠦࡇ࡯↪⚕㧘ᢥᚱౕ╬㧕
㊄㗵
98,040 ౞
130,250 ౞
68,960 ౞
27,50 ౞
ว ⸘
300,000 ౞
㧙㧙
⴫㧝㧚ࠢࠝ࡜ࡓ࠮ࡦࠪࡦࠣㆮવሶߩ✛⤥⩶ᩣߦ߅ߌࠆ଻᦭₸
✛⤥⩶ᩣ
lasI
lasR
rhlI
rhlI
⥃ᐥᩣ (31 ᩣ)
100%
100%
100%
100%
ᴡᎹᩣ (88 ᩣ)
100%
100%
100%
100%
ᴪጯᩣ (35 ᩣ)
100%
100%
100%
100%
⴫㧞㧚࠲ࠗࡊ III ಽᴲᯏ᭴㑐ㅪㆮવሶߩ✛⤥⩶ᩣߦ߅ߌࠆ଻᦭₸
✛⤥⩶ᩣ
Translocator ㆮવሶ
Effecter ㆮવሶ
pcrV
popB
popD
exoT
exoS
exoU
exoY
⥃ᐥᩣ (31 ᩣ)
100%
100%
100%
100%
64.5%
35.5%
87.1%
ᴡᎹᩣ (88 ᩣ)
100%
100%
100%
100%
100%
1.1%
98.9%
ᴪጯᩣ (35 ᩣ)
100%
100%
100%
100%
11.4%
88.6%
100%
㧙㧙

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