微生物气溶胶采集技术的特点及应用

微生物气溶胶是悬浮于空气中粒径差异显著的生物粒子。污水处理、垃圾填埋等污水和固体废弃物的处理过程会产生大量的微生物气溶胶。近年来,随着对微生物气溶胶的不断认识,对其产生、逸散以及危害环境和人体的研究越来越多。在过去的150年,研究者们研发了多种微生物气溶胶采集技术和仪器设备,每种采集技术各有特点和适用条件。本文阐述沉降法、惯性采样法和过滤法3种典型微生物气溶胶采集技术的特点和原理,分析各种采样设备的适用性,为微生物气溶胶的采集和研究提供参考。     

研究集中空调军团菌生物气溶胶的监测方法

目的:调查和研究军团菌生物气溶胶的污染状况及其监测方法。方法:用微生物气溶胶浓缩器与标准生物气溶胶采样器联合采集取样,采集集中空调的冷却水作为水样,检测其中军团菌生物气溶胶的样品数量,并鉴定它们血清型。结果:在水样中可以检测到军团菌生物气溶胶阳性样品。结论:表明集中空调内存在大量的军团菌生物气溶胶。我们应该加强对集中空调内军团菌生物气溶胶的监测和处理,防止其危害人们的生命健康,导致人们换上呼吸道感染疾病。     

废气生物处理设施排放的微生物气溶胶逸散特征、影响因素与暴露风险研究进展

生物处理技术因其具有高效、成本低廉、操作简便、清洁、无二次污染等特点,已被广泛应用于废气处理方面,但微生物气溶胶会作为二次污染物从废气处理设施排放到周围空气中。由于携带和传播有害微生物,微生物气溶胶对人体健康造成潜在危害和风险。废气生物处理设施既是微生物气溶胶的“汇”,也是微生物气溶胶的“源”。本文阐述了废气生物处理设施微生物气溶胶的逸散水平、群落结构和粒径分布特征,分析了其形成原因、主要来源、影响因素和暴露风险,为废气生物处理设施产生的微生物气溶胶的识别和控制技术研究提供科学依据和参考。     

人为源微生物气溶胶的分布特征及风险研究进展

近年来,新型冠状病毒肺炎疫情全球肆虐,引起了公众对于微生物气溶胶潜在风险的极大关注,其中人为源微生物气溶胶潜在的健康危害逐步成为越来越多学者关注的热点之一。本文综述了近年来4类主要人为源微生物气溶胶的研究现状,比较了不同人为源微生物气溶胶的分布特征和微生物组成特性,并探究了影响微生物气溶胶特征的主要因素及其存在的潜在风险。结果表明,畜禽养殖场微生物气溶胶平均浓度最高,其次是污水处理厂和垃圾填埋场,医院最低。从微生物组成特性来说,不同人为源微生物气溶胶中微生物组成与其产生源密切相关;同时,其组成也受其所处环境条件影响。基于以上分析,本文进一步展望了未来人为源微生物气溶胶的主要研究方向,以期为微生物气溶胶控制标准的制定及控制技术的研发奠定基础。     

马桶冲水行为与微生物气溶胶传播

有关卫生间马桶冲水行为可能产生的微生物气溶胶扩散与病原微生物传播的情况一直受到广泛关注,相关研究或报道时有出现。本文意在总结与马桶冲水行为产生的生物气溶胶相关的研究及进展,讨论马桶冲水行为、开关马桶盖冲水行为以及马桶表面等因素在病毒、细菌及寄生虫等病原微生物通过生物气溶胶进行传播的过程中起到的作用。此外,本文还探讨了与马桶相关的微生物气溶胶的产生可能为人体健康带来的潜在健康风险,并就公众健康防护以及未来研究方向提出建议。     

检测病毒气溶胶存活的方法学研究

以钝齿棒状杆菌噬菌体B271血清型为病毒等小颗粒生物粒子的模拟剂,建立了一种适合这类小颗粒生物粒子气溶胶存活研究的方法。本文从该噬菌体耐气溶胶化特性、气溶胶粒谱、用气溶胶示踪剂求算物理衰亡的方法和气溶胶采样回收技术等方面探讨了病毒气溶胶存活研究中的几个关键技术问题,为病毒气溶胶存活研究提供了参考。     

分子生物学在空气微生物气溶胶研究中的应用进展

空气微生物气溶胶的研究方法很多,包括培养计数法、生物发光法、化学发光法、直接镜检法、免疫学方法、分子标志法、生物传感器和分子生物学方法(基因探针和PCR法)。分子生物学方法由于特异性强、操作简便、快速,尤其是最新发展的定量PCR的方法还可以实现对DNA或RNA的绝对定量分析,因此在空气微生物气溶胶的研究中有很好的发展前景。本文对应用PCR和实时定量PCR(QPCR)在空气微生物气溶胶领域的研究进行了综述,指出了传统PCR和QPCR的优缺点,着重对QPCR在空气微生物领域的应用进行了比较全面的分析,指出了应用QPCR检测空气微生物气溶胶还应解决的问题和前景。     

卫生填埋场微生物气溶胶的逸散及潜在风险

随着对微生物气溶胶认识的提高,其产生、来源、扩散及风险研究获得了越来越多的关注。卫生填埋场是微生物气溶胶的重要产生源之一。本文阐述了卫生填埋场气溶胶颗粒中微生物的浓度水平、粒径分布、种群结构,解析了微生物气溶胶的逸散特征及影响因素,介绍了微生物气溶胶对人体健康的潜在风险及评价方法,并展望了未来卫生填埋场逸散微生物的研究趋势及方向,为卫生填埋场微生物气溶胶的控制与削减提供了科学依据和参考。     

废气处理生物滤器微生物生态学研究进展

废气生物处理是一项新兴的气体污染控制技术,已成为当前环境领域的研究热点.本文概述了生物滤器系统(包括生物过滤器系统和生物滴滤器系统)结构及其去除气体污染物的原理,重点阐述了生物滤器微生物的分离、鉴定及其降解特性,微生物丰度、活性与微生物群落结构多样性之间的相关性,运行条件对微生物群落的影响,微生物群落结构时空变化规律,生物膜形成机理和模型等方面的研究进展,并对今后废气处理生物滤器微生物生态学研究方向进行了展望.     

近海潮间带植物及其生境中微生物的特性及药用前景

近海潮间带植物生境的微生物是最富有生物多样性的海洋种群,其生境的独特性决定了该区域微生物较高的药用价值.本文介绍了我国南北方海域近海潮间带植物生境和微生物的特性及药用前景,为该区域微生物的开发利用提供参考.     

Linking the conventional and emerging detection techniques for ambient bioaerosols: a review

Bioaerosols are biologically originated particles present in the atmosphere that can be formed from any process involving biological materials. They comprise of both living and non-living components including organisms, dispersal methods of organisms, and excretions. Bioaerosols such as airborne bacteria, fungal spores, pollen, and others possess diverse characteristics and effects. A large gap exists in the scientific understanding of the overall physical characteristics and measurement of bioaerosols. Consequently, this review aims to devise an appropriate approach to generate more scientific knowledge of bioaerosols. In addition to comparisons and discussions about the various factors affecting bioaerosols, sampling, handling, and the application of various devised analytical techniques, this review offers insight into the current state of bioaerosol research. The review focuses on instrumental and methodical strategies to understand bioaerosol measurement. Numerous studies have investigated conventional methods, advanced methods, and real-time methods that can be applied for bioaerosol monitoring. Each method is different in terms of working principle, characteristics, sensitivity, and efficiency. For the first time, this review explains and compares different methods of conventional, offline, online, and real-time detection methods of bioaerosols based on their working principles, sensitivity, and efficiency on a single platform. This will provide a clear concept and better options for selecting the appropriate method based on the research proposal. Furthermore, recent advances are summarized, and future outlooks are emphasized for bioaerosol identification and categorization. This study also encourages developing affordable and standardized methods to avoid the inter-laboratory and sampling variability to obtain a better understanding and comparison of bioaerosol measurements worldwide. Nevertheless, this work can assist researchers in selecting appropriate methods for bioaerosol measurement and investigation.

     

Bioaerosol biomonitoring: Sampling optimization for molecular microbial ecology

Bioaerosols (or biogenic aerosols) have largely been overlooked by molecular ecologists. However, this is rapidly changing as bioaerosols play key roles in public health, environmental chemistry and the dispersal ecology of microbes. Due to the low environmental concentrations of bioaerosols, collecting sufficient biomass for molecular methods is challenging. Currently, no standardized methods for bioaerosol collection for molecular ecology research exist. Each study requires a process of optimization, which greatly slows the advance of bioaerosol science. Here, we evaluated air filtration and liquid impingement for bioaerosol sampling across a range of environmental conditions. We also investigated the effect of sampling matrices, sample concentration strategies and sampling duration on DNA yield. Air filtration using polycarbonate filters gave the highest recovery, but due to the faster sampling rates possible with impingement, we recommend this method for fine ‐scale temporal/spatial ecological studies. To prevent bias for the recovery of Gram‐positive bacteria, we found that the matrix for impingement should be phosphate‐buffered saline. The optimal method for bioaerosol concentration from the liquid matrix was centrifugation. However, we also present a method using syringe filters for rapid in‐field recovery of bioaerosols from impingement samples, without compromising microbial diversity for high ‐throughput sequencing approaches. Finally, we provide a resource that enables molecular ecologists to select the most appropriate sampling strategy for their specific research question.     

Enhancing Bioaerosol Sampling by Andersen Impactors Using Mineral-Oil-Spread Agar Plate

As a bioaerosol sampling standard, Andersen type impactor is widely used since its invention in 1950s, including the investigation of the anthrax attacks in the United States in 2001. However, its related problems such as impaction and desiccation stress as well as particle bounce have not been solved. Here, we improved its biological collection efficiencies by plating a mineral oil layer (100 µL) onto the agar plate. An Andersen six-stage sampler and a BioStage impactor were tested with mineral-oil-spread agar plates in collecting indoor and outdoor bacterial and fungal aerosols. The effects of sampling times (5, 10 and 20 min) were also studied using the BioStage impactor when sampling environmental bioaerosols as well as aerosolized Bacillus subtilis (G+) and Escherichia coli (G-). In addition, particle bounce reduction by mineral-oil-plate was also investigated using an optical particle counter (OPC). Experimental results revealed that use of mineral-oil-spread agar plate can substantially enhance culturable bioaerosol recoveries by Andersen type impactors (p-values<0.05). The recovery enhancement was shown to depend on bioaerosol size, type, sampling time and environment. In general, more enhancements (extra 20%) were observed for last stage of the Andersen six-stage samplers compared to the BioStage impactor for 10 min sampling. When sampling aerosolized B. subtilis, E. coli and environmental aerosols, the enhancement was shown to increase with increasing sampling time, ranging from 50% increase at 5 min to ∼100% at 20 min. OPC results indicated that use of mineral oil can effectively reduce the particle bounce with an average of 66% for 10 min sampling. Our work suggests that enhancements for fungal aerosols were primarily attributed to the reduced impaction stress, while for bacterial aerosols reduced impaction, desiccation and particle bounce played major roles. The developed technology can readily enhance the agar-based techniques including those high volume portable samplers for bioaerosol monitoring.     

Development of a fungal spore aerosol generator: test with Cladosporium cladosporioides and Penicillium citrinum

As the first step to develop efficient means to control fungal spore bioaerosols, we designed, manufactured, and evaluated a fungal spore aerosol generator. We studied the physical and biological properties of the fungal spore bioaerosols on two common fungal species. The results demonstrated that the fungal spore bioaerosol generator effectively produces fungal spore bioaerosols.     

Comparison of bioaerosol sampling methods in barns housing swine.

The air in livestock buildings contains bioaerosol levels that are sufficiently high to cause adverse health effects in animals and workers. These bioaerosols are complex mixtures of live and dead microorganisms and their products as well as other aeroallergens. The effectiveness of sampling methods used for quantifying the very high concentrations of microorganisms in these environments has not been well studied. To facilitate an accurate assessment of respiratory hazards from viable organisms in agricultural environments, three bioaerosol sampling methods were investigated: the Andersen microbial sampler method (AMS), the all-glass impinger method (AGI), and the Nuclepore filtration-elution method (NFE). These methods were studied in a parallel fashion in 24 swine confinement buildings. Measurements were taken in two seasons with three types of culture media in duplicate to assess total bacteria, gram-negative enteric bacteria, and total fungi. Methods were analyzed for the proportion of samples yielding data within the limits of detection, intraclass reliability, and correlation between methods. For sampling viable bacteria, the AMS had a poor data yield because of overloading and demonstrated weak correlation with the AGI. Conversely, the AGI and NFE gave sufficient numbers of valid data points (90%), yielded high intraclass reliabilities (alpha greater than or equal to 0.92), and were highly correlated with each other (r = 0.86). The AGI and the NFE were suitable methods for assessing bacteria in this environment, but the AMS was not. The AMS was the only method that consistently recovered enteric bacteria (73% data yield). For sampling fungi, the AGI and AMS both yielded sufficient data and all three methods demonstrated high intraclass reliability.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bioaerosols associated with animal production operations

Air emissions from animal housing and manure management operations include a complex mixture of biological, microbial, and inorganic particulates along with odorous volatile compounds. This report highlights the state of current issues, technical knowledge, and remaining challenges to be addressed in evaluating the impacts of airborne microorganisms, dusts, and odorants on animals and workers at animal production facilities and nearby communities. Reports documenting bioaerosol measurements illustrate some of the technical issues related to sample collection, analysis, as well as dispersion and transport to off-farm locations. Approaches to analysis, mitigation and modeling transport are discussed in the context of the risk reduction and management of airborne spread of bioaerosols from animal operations. The need for standardization and validation of bioaerosol collection and analytical techniques for indoor as well as outdoor animal agriculture settings is critical to evaluation of health effects from modern animal production systems that are increasingly situated near communities.     

Application of flow cytometry and fluorescent in situ hybridization for assessment of exposures to airborne bacteria.

Current limitations in the methodology for enumeration and identification of airborne bacteria compromise the precision and accuracy of bioaerosol exposure assessment. In this study, flow cytometry and fluorescent in situ hybridization (FISH) were evaluated for the assessment of exposures to airborne bacteria. Laboratory-generated two-component bioaerosols in exposures chambers and complex native bioaerosols in swine barns were sampled with two types of liquid impingers (all-glass impinger-30 and May 3-stage impinger). Aliquots of collection media were processed and enumerated by a standard culture technique, microscopy, or flow cytometry after nucleic acid staining with 4',6-diamidino-2-phenylindole (DAPI) and identified taxonomically by FISH. DAPI-labeled impinger samples yielded comparable estimates of bioaerosol concentrations when enumerated by microscopy or flow cytometry. The standard culture method underestimated bioaerosol concentrations by 2 orders of magnitude when compared to microscopy or flow cytometry. In the FISH method, aliquots of collection media were incubated with a probe universally complementary to eubacteria, a probe specific for several Pseudomonas species, and a probe complementary to eubacteria for detection of nonspecific binding. With these probes, FISH allowed quantitative identification of Pseudomonas aeruginosa and Escherichia coli bioaerosols in the exposure chamber without measurable nonspecific binding. Impinger samples from the swine barn demonstrated the efficacy of the FISH method for the identification of eubacteria in a complex organic dust. This work demonstrates the potential of emerging molecular techniques to complement traditional methods of bioaerosol exposure assessment.