基于宏基因组测序的星级酒店集中空调系统微生物污染特征

Microbial contamination characteristics of centralized air conditioning systems in star-rated hotels based on metagenomic sequencing

  • 摘要:
    背景 公共场所集中空调通风系统(集通系统)的使用有利于改善室内空气质量,增加人群体感舒适度。但是,集通系统也可能成为室内空气致病微生物污染的潜在载体。
    目的 了解星级酒店集通系统微生物污染特征及病原微生物分布特点,为有效控制其污染提供依据。
    方法 按照WS 394—2012《公共场所集中空调通风系统卫生规范》要求,在2020年9月9日随机选取上海市中心城区2家星级酒店(A和B)的2套集通系统(启用年代分别为2002年、1998年)进行送风中微生物和送风中可吸入颗粒物(PM10)、风管内表面积尘量和微生物、冷却水和冷凝水中嗜肺军团菌等卫生学评价。同时在每套集通系统采集风管内表面样品3件,滤网表面、冷凝水、冷却水样品各1件,2套集通系统共采集12件样品进行Illumina HiSeq宏基因组测序,依据样品类型将其分为3组:风管组、滤网组、水样组。计算α多样性指数(包括Shannon指数、Simpson指数、Chao1指数、ACE指数、良好覆盖指数),反映微生物群落组成特征;通过主成分分析研究3组的β多样性,确定各组样品微生物群落相似程度。
    结果 酒店A集通系统送风中细菌总数、真菌总数最大值分别为1158、344 CFU·m−3,酒店B分别为2000、532 CFU·m−3。两酒店β-溶血性链球菌均未检出,送风中可吸入颗粒物、风管内表面微生物及积尘量、冷却水和冷凝水样品中嗜肺军团菌指标均符合卫生标准要求。Illumina HiSeq宏基因组测序结果表明,12个样本中共测得17322种微生物,其中细菌微生物群占已分类基因的97.31%,其余2.69%来自真菌、病毒和寄生虫。在种水平上,表皮葡萄球菌、假单胞菌、链格孢菌、马拉色菌等为本次调查测得的优势微生物类群。α多样性分析结果表明3组样品的Shannon、Simpson、Chao1、ACE指数值均为风管>滤网>水样。各组样品良好覆盖指数均接近于1。主成分分析表明,两个主成分贡献率分别为19.27%、14.25%,其中,滤网及风管组样品较好地聚为一类。
    结论 2家星级酒店除集通系统送风中微生物污染较严重外,总体卫生状况良好。宏基因组测序发现集通系统微生物类群结构复杂,物种组成包括细菌、真菌、病毒和寄生虫等;不同类型样品物种组成有差别,滤网和风管的样品物种组成接近且以人源性病原微生物为主,提示集通系统潜在的生物安全隐患不容忽视。

     

    Abstract:
    Background The use of heating and ventilation air conditioning (HVAC) in public places is conducive to improving indoor air quality and increasing the users comfort level. However, HVAC may also become potential carriers of indoor airborne microbial contamination.
    Objective To understand the characteristics of microbial pollution and distribution of related pathogenic microorganisms in HVAC of star-rated hotels, and to provide a basis for effective control of such pollution.
    Methods According to the requirements of the Hygienic specification of central air conditioning ventilation system in public buildings (WS 394-2012), two sets of HVAC in two star-rated hotels (A and B, inaugurated in 2002 and 1998, respectively) in the central area of Shanghai were randomly selected on September 9, 2020 for the hygienic evaluation of microorganisms in the air supply and respirable particulate matter (PM10) in the air supply, dust accumulation and microorganisms on the inner surface of the ducts, as well as Legionella pneumophila in cooling water and condensate water. At the same time, 3 samples from the inner surface of ducts, 1 sample from the surface of the filter, 1 sample from the condensate, and 1 sample from the cooling water were collected from each set of HVAC, a total of 12 sample from 2 sets of HVAC, for Illumina HiSeq metagenomic sequencing, and the samples are divided into 3 groups according to their types: duct group, filter group, and water sample group. The α-diversity indices (Shannon index, Simpson index, Chao1 index, ACE index, and goods_coverage index) were calculated to reflect the microbial community composition; and the β-diversity of the three groups were studied by principal component analysis to determine the similarity of the microbial communities.
    Results The maximum total number of bacteria and fungi in the air supply of the HVAC were 1158 CFU·m−3 and 344 CFU·m−3 for Hotel A respectively; and 2000 CFU·m−3 and 532 CFU·m−3 for Botel B respectively. β-hemolytic streptococci were negative in all samples; the respirable particulate matter, microorganisms and dust accumulation on the inner surface of air ducts, Legionella pneumophila IN cooling water and condensate samples all met the standards. The results of Illumina HiSeq sequencing showed that a total of 17322 microorganisms were reported in the 12 samples, with bacterial microbiota accounting for 97.31% of the classified genes and the remaining 2.69% were from fungi, viruses, and parasites. At the species level, Staphylococcus epidermidis, Pseudomonas, Alternaria, and Malassezia were the dominant microbial taxa measured in this survey. The results of α-diversity analysis showed that the values of Shannon index, Simpson index, and Chao1 index for the three groups of samples were duct > filter > water sample. The goods coverage indices of all sample groups s were close to 1. The principal component analysis showed that the contributions of two principal components were 19.27% and 14.25%, respectively, in which the samples of the filter and duct groups were better clustered into one category.
    Conclusion The overall hygiene conditions of the two hotels are good, except for the serious microbial contamination in the air supply of HVAC. Metagenomic sequencing reveals complex microbial communities of HVAC, including bacteria, fungi, viruses, and parasites. The species composition vary by sample groups, particularly the species compositions of the samples from filters and ducts are close and dominated by pathogenic microorganisms of human origin, suggesting that the potential biosafety hazards of HVAC should not be ignored.

     

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