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題名：室內空氣中生物氣膠之特徵與微生物活性之現場快速評估
作者：龔佩怡
作者(外文)：Pei-Yikung
校院名稱：國立成功大學
系所名稱：資源工程學系碩博士班
指導教授：申永輝
溫紹炳
學位類別：博士
出版日期：2012
主題關鍵詞：室內空氣品質；生物氣膠；細菌；真菌；ATP生物冷光法；ATP bioluminescence；Bioaerosol；Microbial activity；Indoor air quality；Environmental hygiene.
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本研究於2009年至2010年間，調查台灣南部地區包含醫療院所、學校、辦公大樓、大賣場、圖書館、車站及戲院等39處具代表性之場所，於調查前預先以簡易直讀式儀器對室內環境進行巡檢(walk through)，初步了解各污染物於空間之分布狀況後，再以環保署公告之標準方法於巡檢出高值區域或有特殊意義之位置進行24小時長時間定點檢測。
結果顯示第一類場所中CO2濃度介於438-1527ppm(法規建議值為600ppm)，超標比例61.1%；CO濃度介於0.55-2.14ppm(法規建議值為2ppm)，超標比例11.1%；O3濃度介於0.005-0.072ppm(法規建議值為0.03ppm)，超標比例27.8%；PM10濃度介於22-116ppm(法規建議值為60ppm)，超標比例44.4%；TVOC濃度介於0.32-4.00ppm(法規建議值為3ppm) ，超標比例16.67%；總真菌數(Total Fungi Counts, TFC)介於62-TNTC CFU/m3(法規建議值為1000CFU/m3)，超標比例33.3%；總細菌數(Total Bacteria Counts, TBC)介於196-4875CFU/m3(法規建議值為500CFU/m3)，超標比例83.3%。第二類場所之CO2濃度介於488-1183ppm (法規建議值為1000ppm)，超標比例9.5%；TVOC濃度介於ND-5.6ppm(法規建議值為3ppm)，超標比例9.5%；TFC介於107-4500CFU/m3(法規建議值為1000CFU/m3)，超標比例19.1%；TBC介於178-4125CFU/m3(法規建議值為1000CFU/m3)，超標比例47.6%。其他HCHO及PM2.5在各類場所中皆符合建議值。
將CO2濃度與TBC進行相關性統計分析，結果顯示相關係數r=0.44，而針對醫療院所之分項統計相關係數r=0.55，皆有中度正相關的表現，由於CO2為人體呼吸作用之代謝產物，因此，可間接說明室內活動人數與細菌數之關聯性。另針對單一醫療院所進行生物氣膠調查，發現室內環境經由適當的清潔消毒可有效降低空氣中TBC，過程中TFC反而有驟增的現象，研判係受到沙塵暴事件及醫院旁農地土壤翻作之影響，經以二氧化氯消毒劑對真菌數較高區域進行投藥後即可獲得明顯改善。另針對高濃度超標場所外氣之真菌樣品進行優勢菌種鑑定，獲得三株絲狀真菌之菌種學名分別為CladosporiumPerangustum、Cladosporiumtenuissimum及Fusariumincarnatum。在台灣，最主要之室內空氣品質汙染問題為CO2、TBC及TFC。
ATP生物冷光技術（ATP bioluminescence technique）能快速偵測微生物活性，較一般傳統微生物分析培養方式來獲取微生物之基本資訊更具有操作簡便、節省成本且無須冗長的培養時間等優點，多年來廣泛被應用在食品、環境、醫療及公衛生等領域。本研究將生物冷光技術應用於偵測空氣中微生物活性，根據場所特性規劃空間偵測點位，再將偵測之微生物活性(相對光量，RLU)藉由繪圖軟體(SURFER 9.0)予圖形化處理，即可呈現微生物活性之分布狀況，經比對偵測場所之空間規劃與擺設狀況，發現微生物活性旺盛區域除了與活動範圍之人數多寡有關外，也涵蓋了室內空氣流動方向、室內植栽、垃圾桶、鞋櫃及候車區座位的陳設等區域；另針對空氣中真菌數超標之醫院，進行消毒劑噴灑前後之殺菌效率測試，也可在短時間內獲知空間之殺菌率，俾便提供公共場所進行空間清潔消毒時，規劃藥劑用量、施放位置及施放時間頻率之參考；本研究偵測空氣中微生物活性時，亦對其他污染物(CO2、CO、O3、PM10、PM2.5、HCHO、TVOC、Temp及RH)進行同步偵測，經相關係數矩陣的彙整，可明顯發現不同之室內環境與場所，各污染物之可能來源與彼此之相關特性亦不盡相同；於現場進行空氣中微生物活性與空氣中總細菌數同步採樣，並將分析結果進行比對，發現即便兩種方法之分析原理與所提供之微生物資訊不同，相關係數(r=0.251)仍有低度正相關之表現，代表現階段掌握之資訊不適於藉由微生物活性指標來推估細菌菌落數，儘管如此，生物冷光技術測得之微生物活性所呈現出之相對意義，仍可提供作為空間微生物密集程度之判斷依據。
空氣中微生物活性偵測技術，數分鐘內即可獲得空氣中微生物活性資訊，無需數日之培養過程，操作簡便、靈敏度高，在短時間內即可得到檢測結果，可提供場所進行篩選調查及消毒改善之依據，因此具有其他微生物檢測方法無可比擬的便捷優勢。
This study investigated indoor air quality (IAQ) at 39 public sites in southern Taiwan including hospitals, schools, office buildings, hypermarkets, libraries, railway stations, theaters, etc. Indoor air quality was preliminarily assessed using handy digital apparatus. Items detected include carbon dioxide (CO2), carbon monoxide (CO), formaldehyde (HCHO), total volatile organic compounds (TVOCs), total bacteria counts (TBC), total fungi counts (TFC), PM10, PM2.5, ozone (O3) and temperature. Based on the results of walk-through detection, the spatial distribution of indoor air contaminants was further measured over a 24 hour period using the EPA standard method. Major indoor air pollutants were found to include CO2, TBC, and TFC. The measured CO2 concentrations ranged between 438 and 1527 ppm, and only 38.9% of them met the Taiwan EPA suggested threshold of 600 ppm. In the schools and hospitals (Category 1), the measured TFC and TBC concentrations ranged from 62 to TNTC CFU/m3 and from 196 to 4875 CFU/m3, respectively. 33% TFC and 83% TBC concentrations exceeded the suggested threshold, and CO2 concentrations were moderately correlated with TBC levels. In a case study of hospital bioaerosols, high TBC and TFC levels were effectively lowered through disinfectant housekeeping as well as ClO2 spray. Three filamentous fungus genera were identified as Cladosporium perangustum, Cladosporium tenuissimum, and Fusarium incarnatum from outdoor samples with high TFC concentrations.
An ATP bioluminescence method was developed for detecting microbial activity in indoor air. This method was compared with the traditional method of collection, culture and count of CFUs. The comparison showed that ATP bioluminescence, expressed as RLUs, was moderately correlated with the entire set of CFU counts (r=0.607), and that correlation improved to r=0.963 (p value〈 0.001) when outlying CFU counts were removed from the calculation. The ATP bioluminescence method was applied at four different sites; a hospital Chinese medicine diagnostic room, a library, a government office, and a railway station lobby. Results showed that microbial activity was far higher in the railway station lobby than at the other three sites and this was seen as a result of the higher volume and density of people in this space. At all four sites, higher microbial activity was linked to indoor plants, garbage cans, shoe racks, and furnished waiting areas. PCA of the data showed that microbial activity in the Chinese medicine diagnostic room was closely related to room temperature and humidity and hence lowering the room humidity can reduce the microbial activity potential here. At all four sites, no correlation was identified between microbial activity and airborne pollutants. The ATP bioluminescence method was applied for the rapid evaluation of room disinfection using chloride dioxide and results showed that twenty minutes after spraying with 100 ppm ClO2, microbial activity was reduced to 38.7% of its original level. ATP bioluminescence is simpler, easier to operate, and more cost-effective than the conventional microbial culture method for evaluating microbial load. The results obtained in this research confirm that the proposed ATP bioluminescence technique is capable of instantaneously detecting microbial activity in an indoor environment. Moreover, the results can be implemented for on-line evaluation of room disinfection efficiency.
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