本文關(guān)鍵詞： 潛艇 微生物氣溶膠 質(zhì)譜 聚合酶鏈反應(yīng)　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:建立適合潛艇艙室的微生物氣溶膠富集技術(shù),跟蹤監(jiān)測潛艇在開放狀態(tài)及封閉狀態(tài)下作業(yè)艙室內(nèi)微生物氣溶膠濃度、菌譜組成及粒徑分布變化,評估其空氣污染程度及潛在的生物風(fēng)險(xiǎn)。方法:比較不同微生物氣溶膠采樣器(LWC-1和安德森)的富集效率,采用固體撞擊式采樣進(jìn)行微生物氣溶膠的富集;采用MALDI-TOF質(zhì)譜技術(shù)對潛艇艙室空氣中可培養(yǎng)微生物進(jìn)行鑒定;應(yīng)用實(shí)時(shí)熒光PCR方法對潛艇艙室空氣中非可培養(yǎng)微生物進(jìn)行檢測;評價(jià)標(biāo)準(zhǔn)依據(jù)《室內(nèi)空氣質(zhì)量標(biāo)準(zhǔn)》(GB/T 1883-2002)。結(jié)果:與LWC-1型和安德森采樣器空氣微生物采樣器相比,安德森采樣器更適于作為本研究的微生物氣溶膠采集器,安德森采樣器的6級不同直徑范圍的孔道可以把空氣中的微生物粒子按照人體呼吸道結(jié)構(gòu)進(jìn)行分級,同時(shí)可以將粒徑作為重要參數(shù)之一與微生物氣溶膠濃度一起評價(jià)空氣微生物污染程度,同時(shí)可進(jìn)行非可培養(yǎng)微生物氣溶膠的富集鑒定;艙室內(nèi)的微生物氣溶膠,航行后濃度比航行前明顯升高(P0.05),不同艙室以及不同級平皿上的菌落數(shù)均增加;長期封閉作業(yè)時(shí),航行前可培養(yǎng)微生物為21個(gè)屬56個(gè)種,革蘭氏陽性球菌和革蘭氏陽性芽孢桿菌占總菌數(shù)為55.11%,革蘭氏陰性桿菌占總菌比例為4.23%,并且均為非發(fā)酵糖革蘭陰性桿菌,航行后為37個(gè)屬80個(gè)種,革蘭陰性桿菌占總菌比例為29.75%,腸桿菌科占革蘭陰性菌的比例為24.4%,航行后革蘭陰性桿菌增加明顯,革蘭陽性球菌和革蘭陽性芽孢桿菌占總菌數(shù)25%,真菌類占總菌比例為24.75%,生物多樣性增加顯著,絲狀真菌和革蘭陰性桿菌明顯增加;航行前后優(yōu)勢細(xì)菌種群發(fā)生明顯改變;FQ-PCR未檢到甲型流感病毒、乙型流感病毒、呼吸道合胞病毒、副流感病毒、腺病毒、嗜肺軍團(tuán)菌、肺炎衣原體、肺炎支原體。結(jié)論:選擇的安德森采樣器適于潛艇狹小空間的微生物氣溶膠富集及構(gòu)成分析,可以同時(shí)使用微生物氣溶膠的粒徑和濃度一同評價(jià)空氣微生物污染程度;航行前后潛艇艙室微生物氣溶膠的濃度及構(gòu)成復(fù)雜性明顯增加,不同艙室以及不同級平板的菌落數(shù)均增加,以條件致病菌增加為主要特征;所分離鑒定的菌種都可以在安德森6級采樣器的各級采樣培養(yǎng)平皿上培養(yǎng)獲得,沒有選擇性和特異性,說明細(xì)菌在空氣中存在的狀態(tài)決定于其所附著的塵埃顆粒大小;長航前后微生物氣溶膠的濃度未超出國家關(guān)于室內(nèi)空氣質(zhì)量的標(biāo)準(zhǔn)要求,未檢出呼吸道病毒、嗜肺軍團(tuán)菌、肺炎衣原體和肺炎支原體,但是從微生物粒徑構(gòu)成及菌譜組成變化考慮,對作業(yè)人員的衛(wèi)生健康仍存在潛在風(fēng)險(xiǎn),應(yīng)采取合理有效的空氣凈化消毒措施,從而保證作業(yè)人員的健康及工作效率。
[Abstract]:Objective: to establish a microorganism aerosol enrichment technique suitable for submarine cabin, and to track and monitor the changes of microorganism aerosol concentration, bacterial spectrum composition and particle size distribution in submarine cabin under open and closed conditions. Methods: the enrichment efficiency of different microbial aerosol samplers LWC-1 and Anderson were compared, and solid impact sampling was used to enrich microbial aerosols. MALDI-TOF mass spectrometry was used to identify culturable microorganisms in submarine cabin air, and real-time fluorescent PCR method was used to detect non-culturable microorganisms in submarine cabin air. Results: compared with LWC-1 and Anderson sampler, Anderson sampler is more suitable to be used as microbial aerosol sampler in this study, and the evaluation standard is based on Indoor Air quality Standard (Indoor Air quality Standard) (GB / T 1883-2002.Results: compared with LWC-1 and Anderson sampler, Andersen sampler is more suitable to be used as microorganism aerosol sampler in this study. Andersen samplers have six levels of holes in different diameters that can classify microbial particles in the air according to the structure of the human respiratory tract. At the same time, the particle size can be used as one of the important parameters to evaluate the degree of microbial pollution in the air together with the concentration of microbial aerosol, and the enrichment and identification of non-culturable microbial aerosol can be carried out. The concentration after navigation was significantly higher than that before navigation, and the number of colonies in different compartments and dishes was increased. During the long period of closed operation, the number of microbes cultured before navigation was 21 genera and 56 species. Gram-positive cocci and Gram-positive Bacillus accounted for 55.11, Gram-negative bacilli accounted for 4.23, and all were non-fermentative gram-negative bacilli. After sailing, there were 37 genera and 80 species. Gram-negative bacilli accounted for 29.75% of the total bacteria, and Enterobacteriaceae accounted for 24.4% of Gram-negative bacteria. Gram-positive cocci and Gram-positive Bacillus accounted for 25% of the total bacteria, fungi accounted for 24.75% of the total bacteria, biodiversity increased significantly, filamentous fungi and Gram-negative bacilli increased significantly. There were significant changes in the dominant bacterial population before and after navigation. No influenza A virus, influenza B virus, respiratory syncytial virus, parainfluenza virus, adenovirus, Legionella pneumophila, Chlamydia pneumoniae were detected by FQ-PCR. Conclusion: the selected Anderson sampler is suitable for enrichment and composition analysis of microorganism aerosol in the narrow space of submarine, and the particle size and concentration of microorganism aerosol can be used to evaluate the degree of air microbial pollution at the same time. The concentration and complexity of microbial aerosol in submarine cabin increased obviously before and after the voyage, and the colony number of different cabin and flat plate increased, with the increase of conditional pathogenic bacteria as the main characteristic. All the bacteria isolated and identified can be cultured on the plate of Anderson 6 sampler without selectivity and specificity, which indicates that the presence of bacteria in the air depends on the size of the dust particles attached to them. Before and after long voyage, the concentration of microbial aerosol did not exceed the national standard on indoor air quality. Respiratory viruses, Legionella pneumophila, Chlamydia pneumoniae and Mycoplasma pneumoniae were not detected. However, considering the change of microbial particle size composition and bacterial spectrum composition, there is still a potential risk to the health and health of the workers, and reasonable and effective air purification and disinfection measures should be taken to ensure the health and working efficiency of the workers.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R84

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