潜艇舱室不同状态微生物气溶胶的富集及调查分析

发布时间：2018-03-01 10:12

本文关键词： 潜艇微生物气溶胶质谱聚合酶链反应　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:建立适合潜艇舱室的微生物气溶胶富集技术,跟踪监测潜艇在开放状态及封闭状态下作业舱室内微生物气溶胶浓度、菌谱组成及粒径分布变化,评估其空气污染程度及潜在的生物风险。方法:比较不同微生物气溶胶采样器(LWC-1和安德森)的富集效率,采用固体撞击式采样进行微生物气溶胶的富集;采用MALDI-TOF质谱技术对潜艇舱室空气中可培养微生物进行鉴定;应用实时荧光PCR方法对潜艇舱室空气中非可培养微生物进行检测;评价标准依据《室内空气质量标准》(GB/T 1883-2002)。结果:与LWC-1型和安德森采样器空气微生物采样器相比,安德森采样器更适于作为本研究的微生物气溶胶采集器,安德森采样器的6级不同直径范围的孔道可以把空气中的微生物粒子按照人体呼吸道结构进行分级,同时可以将粒径作为重要参数之一与微生物气溶胶浓度一起评价空气微生物污染程度,同时可进行非可培养微生物气溶胶的富集鉴定;舱室内的微生物气溶胶,航行后浓度比航行前明显升高(P0.05),不同舱室以及不同级平皿上的菌落数均增加;长期封闭作业时,航行前可培养微生物为21个属56个种,革兰氏阳性球菌和革兰氏阳性芽孢杆菌占总菌数为55.11%,革兰氏阴性杆菌占总菌比例为4.23%,并且均为非发酵糖革兰阴性杆菌,航行后为37个属80个种,革兰阴性杆菌占总菌比例为29.75%,肠杆菌科占革兰阴性菌的比例为24.4%,航行后革兰阴性杆菌增加明显,革兰阳性球菌和革兰阳性芽孢杆菌占总菌数25%,真菌类占总菌比例为24.75%,生物多样性增加显著,丝状真菌和革兰阴性杆菌明显增加;航行前后优势细菌种群发生明显改变;FQ-PCR未检到甲型流感病毒、乙型流感病毒、呼吸道合胞病毒、副流感病毒、腺病毒、嗜肺军团菌、肺炎衣原体、肺炎支原体。结论:选择的安德森采样器适于潜艇狭小空间的微生物气溶胶富集及构成分析,可以同时使用微生物气溶胶的粒径和浓度一同评价空气微生物污染程度;航行前后潜艇舱室微生物气溶胶的浓度及构成复杂性明显增加,不同舱室以及不同级平板的菌落数均增加,以条件致病菌增加为主要特征;所分离鉴定的菌种都可以在安德森6级采样器的各级采样培养平皿上培养获得,没有选择性和特异性,说明细菌在空气中存在的状态决定于其所附着的尘埃颗粒大小;长航前后微生物气溶胶的浓度未超出国家关于室内空气质量的标准要求,未检出呼吸道病毒、嗜肺军团菌、肺炎衣原体和肺炎支原体,但是从微生物粒径构成及菌谱组成变化考虑,对作业人员的卫生健康仍存在潜在风险,应采取合理有效的空气净化消毒措施,从而保证作业人员的健康及工作效率。
[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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R84

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