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研究生:林文海
研究生(外文):Lin, Wen-Hai
論文名稱:真菌氣膠採樣技術之評估
論文名稱(外文):Evaluation of Impingement and Filtration Methods for Fungal Bioaerosol Sampling
指導教授:李芝珊李芝珊引用關係
指導教授(外文):Li Chih-Shan
學位類別:博士
校院名稱:國立臺灣大學
系所名稱:公共衛生研究所
學門:醫藥衛生學門
學類:公共衛生學類
論文種類:學術論文
論文出版年:1998
畢業學年度:86
語文別:中文
論文頁數:200
中文關鍵詞:樣品保存衝擊瓶薄膜濾紙果膠濾紙青黴菌酵母菌
外文關鍵詞:ImpingerNucleporegelatin filterPenicillium citrinumsample storageyeast
相關次數:
  • 被引用被引用:2
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近年來生物氣膠與呼吸道相關症狀被認為具高度相關性。而台灣地區地
處亞熱帶,整年高溫高濕,相當適宜生物氣膠生長。同時在居家環境及作
業環境空氣中均曾觀察到高濃度的生物 氣膠存在,尤其在作業環境中,
被發現空氣中存在著高濃度的微生物。然而目前用於偵測生 物氣膠的採
樣技術面臨許多問題和困難,因此,如何獲取具代表性的生物氣膠樣品仍
是評估 生物氣膠健康危害所要面臨的重要課題。本研究即選定AGI-30衝
擊瓶、Nuclepore濾紙及果 膠濾紙三種較常使用的生物氣膠採樣技術,
於實驗室產生青黴菌孢子及念珠菌細胞,以不同 採樣時間、採樣流量、
樣品保存時間及樣品保存溫度進行效能評估。
實驗結果發現,對青黴菌而言,AGI-30衝擊瓶之採樣效能隨採樣流量及採
樣時間增加而遞 減,Nuclepore薄膜濾紙及果膠濾紙則比較不受採樣流
量及採樣時間影響,且其總回收率高 於AGI-30衝擊瓶。而對念珠菌而
言,AGI-30衝擊瓶之採樣效能於高相對濕度(70%)環境中明顯 優於兩種濾
紙,且受採樣流量及採樣時間之影響不明顯。由上述實驗結果可知,不同
採樣方 法對不同之真菌種類有不同之採樣率能,以濾紙採集青黴菌孢子
可得較高之總回收率,反之,對酵母菌類之生物氣膠而言,則以AGI-30衝
擊瓶之總回收率較高。因此在現場採樣時,需針對主要採樣對象選擇適當
採樣方法,且當選擇總回收率較高之採樣方法 (即以濾紙採集青黴菌孢子
或以AGI-30衝擊瓶採集酵母菌細胞) 時,在本研究選定範圍內之採樣流量
及採樣時間並不會對總回收率有明顯之影響。
由於AGI-30衝擊瓶對青黴菌之採樣效能不佳,因此本研究針對AGI-30的採
樣效率與生物氣膠存活率部份作進一步實驗分析。研究發現物理性收集效
率部份,90%以上之真菌氣懸微粒被收 集於AGI-30衝擊瓶中,沈積於入口
彎管的微粒比例遠低於1%,其餘則自衝擊瓶出口逸出。高流量採樣之收集
效率略高於低流量採樣之收集效率,短時間採樣之收集效率也略高於長時
間採樣之收集效率。就存活率研究發現,被收集的真菌孢子之存活率遠低
於酵母菌細胞,且隨採樣流量及採樣時間之增加而遞減,由本研究結果可
知,此低總回收率主要是因為採集過程使青黴菌孢子存活率降低。
樣品保存對採樣結果之影響部份,實驗結果發現,對青微菌孢子而言,
AGI-30樣品之菌落數隨保存時間增加而減少,且其減少程度與保存溫度無
關,顯示AGI-30樣品宜儘快進行培養,而以濾紙採集之樣品可保存較長時
間。念珠菌細胞部份,AGI-30樣品之菌落數在以4℃保存時可 維持數天之
久,隨後會開始進行出芽生殖,使樣品之細胞總數及菌落數隨保存時間增
加而增加。以25℃保存時,因無低溫度抑制效果,微粒總數及菌落數皆隨
保存時間增加而增加。60分鐘樣品之微粒總數及菌落數會在增殖數天後維
時平穩現象,但5分鐘樣品則在保存七天後仍未見 平穩現象,初步研判此
現象是因念珠菌細胞以採集液中之成份 (如Tween80)為營養來源進行增殖
,而60分鐘樣品因細胞濃度較高,使其細胞濃度較快達到飽和。濾紙樣品
之酵母菌菌落數則隨保存時間增加而減少。由總回收率及保存效果之相關
性發現,高回收率之採樣方法,其樣品可保存較長時間,例如以濾紙採集
之孢子樣品及以衝擊瓶採集之酵母菌樣品,其中以衝擊瓶採集之酵母菌樣
品需冷藏以抑制細胞增殖。反之,總回收率較低之採樣方法,其樣本需儘
速進行培養,以防止菌落數減少。
The influences of sampling time and flow rates as well as
sample storage on total recovery of fungal bioaerosols were
evaluated in the laboratory test chamber by three sampling
methods: AGI-30 all glass impingers, Nucleporefiltration and
elution method, and the gelatin filters. A Pitt-3 generator
anda Collison nebulizer aerosolized spores of Penicillium
citrinum and cells ofCandida famata var. flareri, respectively.
The results demonstrated that total recovery of spores collected
by the AGI-30 impinger was in the range of 4% to 24%.Moreover,
the average totalrecoveries of spores collected by Nuclepore and
gelatin filters were found to bein the rangeof 50% to 75%, and
50% to 90%, respectively.The observed low totalrecovery for
AGI-30 impinger was primary due to the biological stress during
sampling process. The total recovery of fungal sporescollected
in AGI-30impingers became lower as the sampling time and flow
ratesincreased. However,no significant influences of sampling
time and flow rateson the total recovery of both Nuclepore
filtration and gelatin-filter methodswere observed. In summary,
it was found that filtration methods could performmuch better
than impingers for sampling airborne fungal spores. To provide
a quantitative basis for determining the airborne
yeastconcentration, thebioefficiency results demonstrated that
the total recovery ofairborne yeast collectedby the AGI-30
impinger was above 80% at 70% relativehumidity, while
decreasedrapidly as relative humidity decreased beheath 60%.
This might be relatedto undergoing desiccation and losing
culturability before collection for yeastcells following
bioaerosol generation from liquid suspension. The
filtrationoutcome indicated that dehydration effect was
significant and total recoverywas less than 20% for Nuclepore
and gelatin filters. In addition, no significanteffects were
observed regarding sampling time and flow rates for these
threesampling methods. Effects of storage time and temperature
on the colony recovery of airborne fungal samples were evaluated
in the same laboratory test chamber. The resultsdemonstrated
that the bioefficiency of impinger for fungal spores decreased
asstorage time increased whatever sampling time and flow rates
were. In contrast,the yeast cells could survive in the impinger
fluid and even utiliz additive tobud more cells. In addition,
the inhibition effect of refrigerated samples wasobserved.
Therefore, it was suggested that the samples collected by
impingementmethod should be refrigerated and processed as soon
as possible to avoid theloss of spore culturability and increase
of yeast cells. Moreover, the effect of storage time filtration
collection for fungal spores was demonstrated to
beinsignificant. However, yeast recovery from filters was
demonstrated to decreaseas storage time increased. It was
concluded that the recovery would not decrease during storage if
bioefficiencies of the sampling methods were excellent,
forexample, using filters to collect fungal spores or impingers
to collect yeastcells.
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