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.     

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。     

Impaction onto a Glass Slide or Agar versus Impingement into a Liquid for the Collection and Recovery of Airborne Microorganisms

To study impaction versus impingement for the collection and recovery of viable airborne microorganisms, three new bioaerosol samplers have been designed and built. They differ from each other by the medium onto which the bioaerosol particles are collected (glass, agar, and liquid) but have the same inlet and collection geometries and the same sampling flow rate. The bioaerosol concentrations recorded by three different collection techniques have been compared with each other: impaction onto a glass slide, impaction onto an agar medium, and impingement into a liquid. It was found that the particle collection efficiency of agar slide impaction depends on the concentration of agar in the collection medium and on the sampling time, when samples are collected on a nonmoving agar slide. Impingement into a liquid showed anomalous behavior with respect to the sampling flow rate. Optimal sampling conditions in which all three new samplers exhibit the same overall sampling efficiency for nonbiological particles have been established. Inlet and collection efficiencies of about 100% have been achieved for all three devices at a sampling flow rate of 10 liters/min. The new agar slide impactor and the new impinger were then used to study the biological factors affecting the overall sampling efficiency. Laboratory experiments on the total recovery of a typical environmental microorganism, Pseudomonas fluorescens ATCC 13525, showed that both sampling methods, impaction and impingement, provided essentially the same total recovery when relatively nonstressed microorganisms were sampled under optimal sampling conditions. Comparison tests of the newly developed bioaerosol samplers with those commercially available showed that the incorporation of our research findings into the design of the new samplers yields better performance data than data from currently available samplers.     

Quantitative sampling of indoor air biomass by signature lipid biomarker analysis

Exposure to airborne biocontaminants may result in a multitude of health effects and is related to a pronounced increase in adult-onset asthma. Established culture-based procedures for quantifying microbial biomass from airborne environments have severe limitations. Assay of the phospholipid fatty acid (PLFA) components of airborne microorganisms provides a quantitative method to define biomass, community composition and nutritional/physiological activity of the microbial community. By collecting airborne particulate matter from a high volume via filtration, we collected sufficient biomass for quantitative PLFA analysis. Comparing high (filtration) and low (impaction) volume air sampling techniques at 26 locations within the Eastern United States, we determined that PLFA analysis provided a viable alternative to the established but flawed culture-based techniques for measuring airborne microbial biomass and community composition. Compared to the PLFA analysis, the culture techniques underestimated the actual viable airborne biomass present by between one to three orders of magnitude. A case study of a manufacturing plant at which there had been complaints regarding the indoor air quality is presented. Phospholipid fatty acid characterization of the biomass enabled contamination point source determination. In comparison with samples taken outdoors, increases in the relative proportion of trans PLFA, reflecting shifts in the physiological status of viable airborne Gram-negative bacteria, were detected in the indoor air samples at a majority of sampling sites. Received 29 September 1998/ Accepted in revised form 8 January 1999     

微生物气溶胶检测技术的研究进展

生物气溶胶中微生物的种类复杂多样,其采集和鉴定是全面掌握微生物气溶胶生物特性的关键,对防控气溶胶病原体传播十分重要.本文简要综述了微生物气溶胶的生物特性和潜在危害性,介绍了微生物气溶胶的一般采集方法,采集器采集生物气溶胶的基本原理、特点和优缺点.将有助于研究人员根据实验目的在采集低浓度生物气溶胶时选择合适的采样器.本文...     

Detection and enumeration of airborne biocontaminants

The sampling and analysis of airborne microorganisms has received attention in recent years owing to concerns with mold contamination in indoor environments and the threat of bioterrorism. Traditionally, the detection and enumeration of airborne microorganisms has been conducted using light microscopy and/or culture-based methods; however, these analyses are time-consuming, laborious, subjective and lack sensitivity and specificity. The use of molecular methods, such as quantitative polymerase chain reaction amplification, can enhance monitoring strategies by increasing sensitivity and specificity, while decreasing the time required for analysis.     

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.     

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。     

污染土壤微生物群落结构多样性及功能多样性测定方法

土壤微生物在促进土壤质量和植物健康方面发挥着重要的作用,土壤微生物群落结构和组成的多样性及其变化在一定程度上反映了土壤质量.为了更好地了解土壤健康状况,非常有必要发展有效的方法来研究污染土壤微生物的多样性、分布以及行为等.回顾了近年来国内外污染土壤微生物群落结构多样性及功能多样性的测定方法,包括生物化学技术和分子生物学技术,现将它们的原理、优缺点、实用性及其发展动态作一阐述,同时指出结合这两种技术可为微生物群落分析提供一个更全面的、精确的方法.     

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.

     

Temporal and spatial changes in the microbial bioaerosol communities in green-waste composting

In this study, the microbial community within compost, emitted into the airstream, downwind and upwind from a composting facility was characterized and compared through phospholipid fatty acid analysis and 16S rRNA gene analysis using denaturing gradient gel electrophoresis and bar-coded pyrosequencing techniques. All methods used suggested that green-waste composting had a significant impact upon bioaerosol community composition. Daily variations of the on-site airborne community showed how specific site parameters such as compost process activity and meteorological conditions affect bioaerosol communities, although more data are required to qualify and quantify the causes for these variations. A notable feature was the dominance of Pseudomonas in downwind samples, suggesting that this genus can disperse downwind in elevated abundances. Thirty-nine phylotypes were homologous to plant or human phylotypes containing pathogens and were found within compost, on-site and downwind microbial communities. Although the significance of this finding in terms of potential health impact was beyond the scope of this study, it clearly illustrated the potential of molecular techniques to improve our understanding of the impact that green-waste composting emissions may have on the human health.     

The Efficient Method for Simultaneous Monitoring of the Culturable as Well as Nonculturable Airborne Microorganisms

Cultivation-based microbiological methods are a gold standard for monitoring of airborne micro-organisms to determine the occupational exposure levels or transmission paths of a particular infectious agent. Some highly contagious microorganisms are not easily culturable but it is becoming evident that cultivation and molecular methods are complementary and in these cases highly relevant. We report a simple and efficient method for sampling and analyzing airborne bacteria with an impactor-type high-flow-rate portable air sampler, currently used for monitoring culturable bacteria or fungi. A method is reported for extraction of nucleic acids from impacted cells without prior cultivation and using agarose as a sampling matrix. The DNA extraction efficiency was determined in spiked samples and, samples taken from a wastewater treatment plant and an alpine area. The abundance, diversity and quantity of total bacteria were analysed by a quantitative polymerase chain reaction, and by construction and analysis of clone libraries. The method does not interfere with downstream PCR analysis and can cover the gap between traditional culture and molecular techniques of bioaerosol monitoring.     

Microbial contamination of storerooms at the Auschwitz-Birkenau Museum

Biodeterioration of heritage collections caused by microorganisms is a worldwide problem. To avoid degradation caused by biological contaminants transported into the indoor environment by air, proper bioaerosol protection is required. The aim of this study was to assess the level of microbial contamination of the Auschwitz-Birkenau Museum collection based on qualitative and quantitative analyses of bacteria and fungi isolated from the atmosphere and settled dust of museum storerooms. The obtained results demonstrated that a correctly operated air-conditioning system and limiting the number of visitors in the studied storerooms can significantly inhibit microbial contamination of the air and decrease deposition of bacterial and fungal particulates on exhibit surfaces. The performed analyses confirm that an aerobiological assessment of museum premises is a useful tool in their hygienic evaluation and, if necessary, in decision-making regarding interventions to minimize biological decay of collections.     

Use of solid-phase PCR for enhanced detection of airborne microorganisms.

The solid-phase PCR (SP-PCR) was compared with a culture-based technique for the detection of aerosolized Escherichia coli DH1. Results with SP-PCR showed an increase in detection sensitivity over that of culture methods. Therefore, SP-PCR may be useful for the detection of airborne microorganisms which may be nonculturable because of aerosolization or sampling stress.     

Sequencing-Based Analysis of the Bacterial and Fungal Composition of Kefir Grains and Milks from Multiple Sources

Kefir is a fermented milk-based beverage to which a number of health-promoting properties have been attributed. The microbes responsible for the fermentation of milk to produce kefir consist of a complex association of bacteria and yeasts, bound within a polysaccharide matrix, known as the kefir grain. The consistency of this microbial population, and that present in the resultant beverage, has been the subject of a number of previous, almost exclusively culture-based, studies which have indicated differences depending on geographical location and culture conditions. However, culture-based identification studies are limited by virtue of only detecting species with the ability to grow on the specific medium used and thus culture-independent, molecular-based techniques offer the potential for a more comprehensive analysis of such communities. Here we describe a detailed investigation of the microbial population, both bacterial and fungal, of kefir, using high-throughput sequencing to analyse 25 kefir milks and associated grains sourced from 8 geographically distinct regions. This is the first occasion that this technology has been employed to investigate the fungal component of these populations or to reveal the microbial composition of such an extensive number of kefir grains or milks. As a result several genera and species not previously identified in kefir were revealed. Our analysis shows that the bacterial populations in kefir are dominated by 2 phyla, the Firmicutes and the Proteobacteria. It was also established that the fungal populations of kefir were dominated by the genera Kazachstania, Kluyveromyces and Naumovozyma, but that a variable sub-dominant population also exists.     

Effect of electrical charges and fields on injury and viability of airborne bacteria

In this study, the effects of the electric charges and fields on the viability of airborne microorganisms were investigated. The electric charges of different magnitude and polarity were imparted on airborne microbial cells by a means of induction charging. The airborne microorganisms carrying different electric charge levels were then extracted by an electric mobility analyzer and collected using a microbial sampler. It was found that the viability of Pseudomonas fluorescens bacteria, used as a model for sensitive bacteria, carrying a net charge from 4100 negative to 30 positive elementary charges ranged between 40% and 60%; the viability of the cells carrying >2700 positive charges was below 1.5%. In contrast, the viability of the stress-resistant spores of Bacillus subtilis var. niger (used as simulant of anthrax-causing Bacillus anthracis spores when testing bioaerosol sensors in various studies), was not affected by the amount of electric charges on the spores. Because bacterial cells depend on their membrane potential for basic metabolic activities, drastic changes occurring in the membrane potential during aerosolization and the local electric fields induced by the imposed charges appeared to affect the sensitive cells' viability. These findings facilitate applications of electric charging for environmental control purposes involving sterilization of bacterial cells by imposing high electric charges on them. The findings from this study can also be used in the development of new bioaerosol sampling methods based on electrostatic principles.     

Field Evaluation of Personal Sampling Methods for Multiple Bioaerosols

Ambient bioaerosols are ubiquitous in the daily environment and can affect health in various ways. However, few studies have been conducted to comprehensively evaluate personal bioaerosol exposure in occupational and indoor environments because of the complex composition of bioaerosols and the lack of standardized sampling/analysis methods. We conducted a study to determine the most efficient collection/analysis method for the personal exposure assessment of multiple bioaerosols. The sampling efficiencies of three filters and four samplers were compared. According to our results, polycarbonate (PC) filters had the highest relative efficiency, particularly for bacteria. Side-by-side sampling was conducted to evaluate the three filter samplers (with PC filters) and the NIOSH Personal Bioaerosol Cyclone Sampler. According to the results, the Button Aerosol Sampler and the IOM Inhalable Dust Sampler had the highest relative efficiencies for fungi and bacteria, followed by the NIOSH sampler. Personal sampling was performed in a pig farm to assess occupational bioaerosol exposure and to evaluate the sampling/analysis methods. The Button and IOM samplers yielded a similar performance for personal bioaerosol sampling at the pig farm. However, the Button sampler is more likely to be clogged at high airborne dust concentrations because of its higher flow rate (4 L/min). Therefore, the IOM sampler is a more appropriate choice for performing personal sampling in environments with high dust levels. In summary, the Button and IOM samplers with PC filters are efficient sampling/analysis methods for the personal exposure assessment of multiple bioaerosols.     

Using a bioaerosol personal sampler in combination with real-time PCR analysis for rapid detection of airborne viruses

We have recently developed a new personal sampler and demonstrated its feasibility for detection of viable airborne microorganisms including bacteria, fungi and viruses. To accelerate the time-consuming analytical procedure involving 2-5 days of biological testing, we employed a real-time PCR protocol in conjunction with the personal sampler for collection of airborne viruses. The advantage of this approach is that if the presence of a particular pathogen in the air is detected by the PCR, the remaining collecting liquid can be further analysed by more time-consuming biological methods to estimate the number of airborne infectious/live microorganisms. As sampling of bioaerosols in natural environments is likely to be associated with substantial contamination by a range of microorganisms commonly existing in an ambient air, an investigation of the specificity of detection by targeted PCR analysis is required. Here we present the results of the study on the detection of Influenza virus in the ambient air contaminated with high concentrations of bacteria and fungi using real-time PCR protocol. The combined sampling PCR detection method was found to be fully feasible for the rapid ( approximately 2.5 h) and highly specific (no cross-reactivity) identification of the labile airborne virus in the air containing elevated concentrations of other microorganisms.     

Recent developments in monitoring and modelling airborne pollen,a review

Abstract

Public awareness of the rising importance of allergies and other respiratory diseases has led to increased scientific effort to accurately and rapidly monitor and predict pollen, fungal spores and other bioaerosols in our atmosphere. An important driving force for the increased social and scientific concern is the realisation that climate change will increasingly have an impact on worldwide bioaerosol distributions and subsequent human health. In this review we examine new developments in monitoring of atmospheric pollen as well as observation and source-orientated modelling techniques. The results of a Scopus® search for scientific publications conducted with the terms ‘Pollen allergy’ and ‘Pollen forecast’ included in the title, abstract or keywords show that the number of such articles published has increased year on year. The 12 most important allergenic pollen taxa in Europe as defined by COST Action ES0603 were ranked in terms of the most ‘popular’ for model-based forecasting and for forecasting method used. Betula, Poaceae and Ambrosia are the most forecast taxa. Traditional regression and phenological models (including temperature sum and chilling models) are the most used modelling methods, but it is notable that there are a large number of new modelling techniques being explored. In particular, it appears that Machine Learning techniques have become more popular and led to better results than more traditional observation-orientated models such as regression and time-series analyses.     

Comparison of aerosol and bioaerosol collection on air filters

Air filters efficiency is usually determined by non-biological test aerosols, such as potassium chloride particles, Arizona dust or di-ethyl-hexyl-sebacate (DEHS) oily liquid. This research was undertaken to asses, if application of non-biological aerosols reflects air filters capacity to collect particles of biological origin. The collection efficiency for non-biological aerosol was tested with the PALAS set and ISO Fine Test Dust. Flow rate during the filtration process was 720 l/h, and particles size ranged 0.246–17.165 μm. The upstream and downstream concentration of the aerosol was measured with a laser particle counter PCS-2010. Tested bioaerosol contained 4 bacterial strains of different shape and size: Micrococcus luteus, Micrococcus varians, Pseudomonas putida and Bacillus subtilis. Number of the biological particles was estimated with a culture-based method. Results obtained with bioaerosol did not confirmed 100% filters efficiency noted for the mineral test dust of the same aerodynamic diameter. Maximum efficiency tested with bacterial cells was 99.8%. Additionally, cells reemission from filters into air was also studied. Bioaerosol contained 3 bacterial strains: Micrococcus varians, Pseudomonas putida and Bacillus subtilis. It was proved that the highest intensity of the reemission process was during the first 5 min. and reached maximum 0.63% of total number of bacteria retained in filters. Spherical cells adhered stronger to the filter fibres than cylindrical ones. It was concluded that non-biological aerosol containing particles of the same shape and surface characteristics (like DEHS spherical particles) can not give representative results for all particles present in the filtered air.