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1.
Microorganism concentrations and viability can be better understood and clarified by using both culture and non-culture methods. Here, using pure suspensions of E. coli, three non-culture methods, namely, flow cytometry (FCM), epifluorescence microscopy (EFM), and real-time quantitative polymerase chain reaction (real-time qPCR), were compared with a traditional culture-based method. Using fluorocome-labeling methods with FCM and EFM applications, acridine orange (AO) and propidium iodide (PI) dyes were used to determine the total cell concentration and microorganism viability, respectively. The results indicated that total cell concentrations determined using FCM were statistically higher (2.62-4.94 times) than those determined using EFM. The difference might be due to cell losses induced by extensive preparations needed for EFM. In addition, EFM and FCM were highly associated for both the total cell concentration and viability. FCM-measured viability was the highest, whereas the culture-measured viability was the lowest. Furthermore, DNA concentrations measured by real-time qPCR with gene probe were highly associated with the total number concentrations measured by either the EFM or FCM. In summary, the three non-culture methods compared here could provide rapid and accurate information about microorganism concentrations and viabilities.  相似文献   

2.
Increasing evidences show that inhalation of indoor bioaerosols has caused numerous adverse health effects and diseases. However, the bioaerosol size distribution, composition, and concentration level, representing different inhalation risks, could vary with different living environments. The six-stage Andersen sampler is designed to simulate the sampling of different human lung regions. Here, the sampler was used in investigating the bioaerosol exposure in six different environments (student dorm, hospital, laboratory, hotel room, dining hall, and outdoor environment) in Beijing. During the sampling, the Andersen sampler was operated for 30 min for each sample, and three independent experiments were performed for each of the environments. The air samples collected onto each of the six stages of the sampler were incubated on agar plates directly at 26 °C, and the colony forming units (CFU) were manually counted and statistically corrected. In addition, the developed CFUs were washed off the agar plates and subjected to polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) for diversity analysis. Results revealed that for most environments investigated, the culturable bacterial aerosol concentrations were higher than those of culturable fungal aerosols. The culturable bacterial and fungal aerosol fractions, concentration, size distribution, and diversity were shown to vary significantly with the sampling environments. PCR-DGGE analysis indicated that different environments had different culturable bacterial aerosol compositions as revealed by distinct gel band patterns. For most environments tested, larger (>3 μm) culturable bacterial aerosols with a skewed size distribution were shown to prevail, accounting for more than 60 %, while for culturable fungal aerosols with a normal size distribution, those 2.1–4.7 μm dominated, accounting for 20–40 %. Alternaria, Cladosporium, Chaetomium, and Aspergillus were found abundant in most environments studied here. Viable microbial load per unit of particulate matter was also shown to vary significantly with the sampling environments. The results from this study suggested that different environments even with similar levels of total microbial cuturable aerosol concentrations could present different inhalation risks due to different bioaerosol particle size distribution and composition. This work fills literature gaps regarding bioaerosol size and composition-based exposure risks in different human dwellings in contrast to a vast body of total bioaerosol levels.  相似文献   

3.
Indoor air quality in health care facilities is a major public health concern, particularly for immunocompromised patients who may be exposed to microbiological contaminants such as molds, mycotoxins, endotoxins, and (1,3)-ß-D-glucans. Over 2 years, bioaerosols were collected on a monthly basis in a cancer treatment center (Centre F. Baclesse, Normandy, France), characterized from areas where there was no any particular air treatment. Results showed the complexity of mycoflora in bioaerosols with more than 100 fungal species identified. A list of major strains in hospital environments could be put forward due to the frequency, the concentration level, and/or the capacity to produce mycotoxins in vitro: Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus versicolor, Cladosporium herbarum, Purpureocillium lilacinum, and Penicillium brevicompactum. The mean levels of viable airborne fungal particles were less than 30.530 CFU per m3 of air and were correlated to the total number of 0.30 to 20 μm particles. Seasonal variations were observed with fungal particle peaks during the summer and autumn. Statistical analysis showed that airborne fungal particle levels depended on the relative humidity level which could be a useful indicator of fungal contamination. Finally, the exposure to airborne mycotoxins was very low (only 3 positive samples), and no mutagenic activity was found in bioaerosols. Nevertheless, some fungal strains such as Aspergillus versicolor or Penicillium brevicompactum showed toxigenic potential in vitro.  相似文献   

4.
In this paper, copper ferrite nanoparticle (CFN) was synthesized, modified by cetyl trimethylammonium bromide, and characterized. Dye removal ability of the surface modified copper ferrite nanoparticle (SMCFN) from single system was investigated. The physical characteristics of SMCFN were studied using Fourier transform infrared, scanning electron microscopy, and X-ray diffraction. Acid Blue 92, Direct Green 6, Direct Red 23, and Direct Red 80 were used as model compounds. The effect of operational parameters (surfactant concentration, adsorbent dosage, dye concentration, and pH) on dye removal was evaluated. Response surface methodology (RSM) was used for the analysis of the dye removal data. The experimental checking in these optimal conditions confirms good agreements with RSM results. The results showed that the SMCFN being a magnetic adsorbent might be a suitable alternative to remove dyes from colored aqueous solutions.  相似文献   

5.
Recently, we reported the outcomes of feasibility studies of a technological approach allowing rapid detection of a wide range of bioaerosols by combining a personal bioaerosol sampler with a real-time PCR technology. The protocol was found suitable for detection of targeted microorganisms within relatively short time periods. Considering the crucial importance of the PCR procedure quality control, the current paper reports the results of the development of an internally controlled PCR system for utilization by the above technology. The suggested strategy is based on utilization of only two fluorescent dyes, which are used respectively for target and internal amplification control (IAC) DNA amplification. A bacteriophage T4 and recombinant phage fd (M13) were used in this research as target and IAC, respectively. The constructed IAC was added directly to the collection liquid of the personal bioaerosol sampler enabling quality control to be present throughout the entire sampling-analysis procedures. For performance evaluation, serial ten-fold dilutions of T4 phage were aerosolized and sampled over a 10 minutes time period. The results showed that T4 phage could be reliably detected at the concentration of around 200 PFU per litre of air over the 10 minutes sampling period. The developed PCR assay demonstrated high specificity and no cross reaction. It is concluded that the recombinant phage fd is suitable for utilization as an internal control enabling to significantly minimize false negative results for bioaerosol detection procedures.  相似文献   

6.
Traditional methods of assessing fungal exposure have been confounded by a number of limiting variables. The recent utilization of molecular methods such as internal transcribed spacer (ITS) sequencing of ribosomal RNA genes has provided improved insight into the diversity of fungal bioaerosols in indoor, outdoor and occupational environments. However, ITS analyses may also be confounded by a number of methodological limitations. In this study, we have optimized this technology for use in occupational or environmental studies. Three commonly used DNA extraction methodologies (UltraClean Soil kit, High Pure PCR Template kit, and EluQuik/DNeasy kit) were compared in terms of sensitivity and susceptibility to PCR inhibitors in dust for three common fungal bioaerosols, Aspergillus versicolor, Rhizopus microsporus and Wallemia sebi. Environmental dust samples were then studied using each extraction methodology and results were compared to viable culture data. The extraction methods differed in terms of their ability to efficiently extract DNA from particular species of fungi (e.g. Aspergillus versicolor). In addition, the ability to remove PCR inhibitors from dust samples was most effective using the soil DNA extraction kit. The species composition varied greatly between ITS clone libraries generated with the different DNA extraction kits. However, compared to viable culture data, ITS clone libraries included additional fungal species that are incapable of growth on solid culture medium. Collectively, our data indicated that DNA extraction methodologies used in ITS sequencing studies of occupational or environmental dust samples can greatly influence the fungal species that are detected.  相似文献   

7.
Personal aerosol samplers are widely used to monitor human exposure to airborne materials. For bioaerosols, interest is growing in analyzing samples using molecular and immunological techniques. This paper presents a personal sampler that uses a two-stage cyclone to collect bioaerosols into disposable 1.5 ml Eppendorf-type microcentrifuge tubes. Samples can be processed in the tubes for polymerase chain reaction (PCR) or immunoassays, and the use of multiple stages fractionates aerosol particles by aerodynamic diameter. The sampler was tested using fluorescent microspheres and aerosolized fungal spores. The sampler had first and second stage cut-off diameters of 2.6 microm and 1.6 microm at 2 l min(-1)(geometric standard deviation, GSD = 1.45 and 1.75), and 1.8 microm and 1 microm at 3.5 l min(-1)(GSD = 1.42 and 1.55). The sampler aspiration efficiency was >or=98% at both flow rates for particles with aerodynamic diameters of 3.1 microm or less. For 6.2 microm particles, the aspiration efficiency was 89% at 2 l min(-1) and 96% at 3.5 l min(-1). At 3.5 l min(-1), the sampler collected 92% of aerosolized Aspergillus versicolor and Penicillium chrysogenum spores inside the two microcentrifuge tubes, with less than 0.4% of the spores collecting on the back-up filter. The design and techniques given here are suitable for personal bioaerosol sampling, and could also be adapted to design larger aerosol samplers for longer-term atmospheric and indoor air quality sampling.  相似文献   

8.
Today's large-scale poultry production is often accompanied by high concentrations of airborne microorganisms at working places. However, the microbial communities in those bioaerosols are rarely characterised. In this study, we investigated the bacterial population in bioaerosols from a duck hatchery by both cultivation based and molecular methods and compared the results. Depending on used media, concentrations of airborne culturable bacteria varied between 6 × 10(1) and 7 × 10(6) CFU per m(3) air. The corresponding total cell count of DAPI stained cells was 2 × 10(7) cells per m(3) air. 16S rRNA gene analyses of bacterial isolates and clone libraries revealed a low species richness in hatcheries air, respectively. More than 50% of bacterial isolates were phylogenetically most closely related to bacterial species of the risk group 2 (German TRBA). The sequence composition in clone libraries supported the result of cultivation based approaches, whereby sequences assigned to Staphylococcus, Acinetobacter and Enterococcus are the most common. The high concentration of airborne bacteria which are most closely related to species of potential health risk requires further detailed investigations for these bacterial species.  相似文献   

9.
Aerosols of water contaminated with Legionella bacteria constitute the only mode of exposure for humans. However, the prevention strategy against this pathogenic bacteria risk is managed through the survey of water contamination. No relationship linked the Legionella bacteria water concentration and their airborne abundance. Therefore, new approaches in the field of the metrological aspects of Legionella bioaerosols are required. This study was aimed at testing the main principles for bioaerosol collection (solid impaction, liquid impingement and filtration) and the in situ hybridization (FISH) method, both in laboratory and field assays, with the intention of applying such methodologies for airborne Legionella bacteria detection while showering. An aerosolization chamber was developed to generate controlled and reproducible L. pneumophila aerosols. This tool allowed the identification of the liquid impingement method as the most appropriate one for collecting airborne Legionella bacteria. The culturable fraction of airborne L. pneumophila recovered with the liquid impingement principle was 4 and 700 times higher compared to the impaction and filtration techniques, respectively. Moreover, the concentrations of airborne L. pneumophila in the impinger fluid were on average 7.0 x 10(5) FISH-cells m(-3) air with the fluorescent in situ hybridization (FISH) method versus 9.0 x 10(4) CFU m(-3) air with the culture method. These results, recorded under well-controlled conditions, were confirmed during the field experiments performed on aerosols generated by hot water showers in health institutions. This new approach may provide a more accurate characterization of aerobiocontamination by Legionella bacteria.  相似文献   

10.
Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml(-1) in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml(-1) in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml(-1) of four real-life dust samples at p-value >0.05 using 2 x 10(4) spores for conventional PCR and 2 x 10(5) spores for real-time PCR. However, samples containing >0.2 mg ml(-1) real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments.  相似文献   

11.
Cladosporium is one of the most common airborne molds found in indoor and outdoor environments. Cladosporium spores are important aeroallergens, and prolonged exposure to elevated spore concentrations can provoke chronic allergy and asthma. To accurately quantify the levels of Cladosporium in indoor and outdoor environments, two real-time PCR systems were developed in this study. The two real-time PCR systems are highly specific and sensitive for Cladosporium detection even in a high background of other fungal DNAs. These methods were employed to quantify Cladosporium in aerosols of five different indoor environments. The investigation revealed a high spore concentration of Cladosporium (10(7) m(-3)) in a cow barn that accounted for 28-44% of the airborne fungal propagules. In a countryside house that uses firewood for heating and in a paper and pulp factory, Cladosporium was detected at 10(4) spores m(-3), which accounted for 2-6% of the fungal propagules in the aerosols. The concentrations of Cladosporium in these three indoor environments far exceeded the medical borderline level (3000 spores m(-3)). In a power station and a fruit and vegetable storage, Cladosporium was found to be a minor component in the aerosols, accounted for 0.01-0.1% of the total fungal propagules. These results showed that monitoring Cladosporium in indoor environments is more important than in outdoor environments from the public health point of view. Cladosporium may not be the dominant fungi in some indoor environments, but its concentration could still be exceeding the threshold value for clinical significance. The methods developed in this study could facilitate accurate detection and quantification of Cladosporium for public health related risk assessment.  相似文献   

12.
Airborne bacteria emissions from oxidation ditch with rotating aeration brushes were investigated in a municipal wastewater treatment plant in Beijing, China. Microbial samples were collected at different distances from the rotating brushes, different heights above the water surface, and different operation state over a 3-month period (April, May, and June) in order to estimate the seasonal variation and site-related distribution characteristics of the microorganisms present. The concentration of bacterial aerosol was analyzed by culture methods, while their dominant species, genetic structure and diversity were assayed using bio-molecular tools. Results showed that total microbial concentrations were highest in June and lowest in April. The mechanical rotation caused remarkable variation in concentration and diversity of culturable airborne bacteria before and after the rotating brushes. The highest concentration was observed near the rotating brushes (931?±?129–3,952?±?730 CFU/m3), with concentration decreasing as distance and height increased. Bacterial community polymerase chain reaction and denaturing gradient gel electrophoresis indicated that diversity decreased gradually with increasing height above the water surface but remained relatively constant at the same height. All dominant bacteria identified by DNA sequence analysis belonged to Firmicutes. Pathogenic species such as Moraxella nonliquefaciens and Flavobacterium odoratum were isolated from the bioaerosols. Due to the serious health risks involved, exposure of sewage workers to airborne microorganisms caused by brush aerators should be monitored and controlled.  相似文献   

13.
近年来,空气污染已成为长三角地区最关键的环境问题之一,气溶胶颗粒物(PM)是最主要的污染物之一.生物气溶胶作为颗粒物的重要组成部分,可能对空气质量和人体健康产生不利影响.利用高通量测序方法研究了江苏省泰州2019年11月至2020年1月期间发生沙尘和霾污染时生物气溶胶中细菌群落结构组成特征.结果表明:冬季沙尘天和霾天气...  相似文献   

14.
沈阳市室内空气细菌与真菌粒子的关系   总被引:3,自引:0,他引:3  
用ANDERSEN生物粒子采样器和平皿沉降法观测了室内空气细菌和真菌粒子浓度、浓度分布、粒度分布、粒数中值直径和沉降量.结果表明,室内空气细菌粒子浓度为真菌粒子浓度的2.5倍,细菌粒数中值直径为真菌粒数中值直径的1.4倍,细菌粒子沉降量为真菌粒子沉降量的5,3倍。<8.2μm的空气细菌和真菌粒子浓度差别不大,而>8.2μm的空气细苗比真苗粒子浓度高4.1倍.<8.2μm的可吸入真菌粒数百分比大于<8.2μm的细菌粒数百分比.在一天的7:00时~22:00时,室内空气细菌和真菌粒子的浓度、沉降量均呈双峰变化;细苗和真菌粒子浓度的相关系数为0.822,细菌和真菌粒子沉降量的相关系数为0.896,均呈明显的正相关关系;细菌和真菌粒数中值直径的相关系数为-0.096,相关关系不大。  相似文献   

15.
Personal exposures to viable fungi and bacteria were compared with the concentrations being assessed by stationary samplers in home and workplace microenvironments. A random sample of 81 elementary school teachers in eastern Finland performed two 24-hour measurement periods in wintertime. Concentrations and prevalences of viable fungi and bacteria on the collection filters were determined by cultivation method. The geometric mean concentration was 3-12 cfu m(-3) for total viable fungi, 0.6-3.7 cfu m(-3) for Penicillium and mainly under 1 cfu m(-3) for other fungi. The samples with higher fungal concentrations also had higher diversity of fungi than samples with lower concentrations. The total number of fungal genera recovered was 39 for personal, 34 for home and 23 for work samples. The variation in concentration of Penicillium explained even 25-95% of the variations of total fungal concentration in personal exposure, home and workplace environments. There was an association between personal exposure and home concentration of viable fungi and between personal exposure and home and work concentrations of viable bacteria. Personal exposure and home concentrations of fungi were higher in rural areas than in urban areas. Our results also indicate that presence of a certain fungus in a microenvironment does not necessarily mean similar findings in personal exposure samples.  相似文献   

16.
This research studied the application of fish bone as a low-cost adsorbent for the removal of two basic dyes (C.I. Basic Blue 41 and C.I. Basic Yellow 28) from textile wastewaters. The surface morphology and functional groups of the fish bone was studied by scanning electron microscopy and Fourier transform infrared, respectively. The experiments were designed in two methods: classical method and response surface methodology (RSM). The effect of operating parameters including pH, initial dye concentration, adsorbent dosage, and inorganic salts was investigated by both methods. The optimum conditions to satisfy 75 % of dye removal for both dyes were predicted by RSM method and the results were very close to the experimental values. So, it was concluded that the fish bone can be used as a low-cost adsorbent for dye removal from effluents.  相似文献   

17.
Airborne transmission of bacterial pathogens from point sources (e.g., ranches, dairy waste treatment facilities) to areas of food production (farms) has been suspected. Determining the incidence, transport and viability of extremely low levels of pathogens require collection of high volumes of air and characterization of live bacteria from aerosols. We monitored the numbers of culturable bacteria in urban aerosols on 21 separate days during a 9 month period using high volume cyclonic samplers at an elevation of 6 m above ground level. Culturable bacteria in aerosols fluctuated from 3 CFU to 6 million CFU/L of air per hour and correlated significantly with changes in seasonal temperatures, but not with humidity or wind speed. Concentrations of viable bacteria determined by fluorescence staining and flow cytometry correlated significantly with culturable bacteria. Members of the phylum Proteobacteria constituted 98% of the bacterial community, which was characterized using 16S rRNA gene sequencing using DNA from aerosols. Aquabacterium sp., previously characterized from aquatic environments, represented 63% of all clones and the second most common were Burkholderia sp; these are ubiquitous in nature and some are potential human pathogens. Whole genome amplification prior to sequencing resulted in a substantial decrease in species diversity compared to characterizing culturable bacteria sorted by flow cytometry based on scatter signals. Although 27 isolated colonies were characterized, we were able to culture 38% of bacteria characterized by sequencing. The whole genome amplification method amplified DNA preferentially from Phyllobacterium myrsinacearum, a minor member of the bacterial communities, whereas Variovorax paradoxus dominated the cultured organisms.  相似文献   

18.
In the present study, organismic-level acute toxicology profile of three reactive azo dyes, viz. Reactive Blue 221, Reactive Red 195, and Reactive Yellow 145, was investigated, by using bacterial (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis), fungal (Trichoderma asperellum, Aspergillus flavus, Fusarium fujikuroi, and Rhizoctonia solani), plant (Raphanus sativus, Triticum aestivum, Sorghum bicolor, and Phaseolus mungo), and aquatic (Artemia salina and Daphnia magna) specimens. Microbial test organisms (all the six bacteria and two fungi, i.e., T. asperellum and A. flavus) and D. magna were found to be relatively more sensitive towards the reactive azo dyes and their mixture, as the EC50 values were in the range of 80–330, 135–360, and 108–242 ppm for bacteria, fungi, and D. magna, respectively (but the effect was not acutely toxic). Moreover, the effect of dye mixture was comparable tothe individual dyes in almost all the tested microbial specimens. For plant seeds, the dye mixture was found to be relatively more inhibitory towards T. aestivum and R. sativus than the individual dyes. For S. bicolor and P. mungo seeds, the effect of the dye mixture was almost identical to the individual dyes. However, in all cases, EC50 values were in the range of 950–3500 ppm, which indicates a non-toxic effect on plant seed germination potential. Likewise, the dyes and their mixture were not acutely toxic for Artemia salina larvae (more sensitive to the dye mixture) and Daphnia magna neonates (EC50, 516–950 and 108–242 ppm, respectively).  相似文献   

19.
The aim of this study was to compare the personal exposure to particles and bioaerosols with that measured by stationary samplers in the main microenvironments, i.e., the home and the workplace. A random sample of 81 elementary school teachers was selected from the 823 teachers working for two councils in eastern Finland for the winter time measurement period. Bioaerosol and other particles were collected on filters by button samplers using personal sampling and microenvironmental measurements in homes and workplaces. The 24-hour sampling period was repeated twice for each teacher. Particle mass, absorption coefficient of the filter and the concentration of viable and total microorganisms were analyzed from each filter. In this paper, the study design, quality assurance principles and results of particle and bioaerosol exposure are described. The results show that particle mass concentrations, absorption coefficient and fungi were higher in personal exposure samples than in home and workplace samples. Furthermore, these concentrations were usually lower in the home than in the workplace. Bacterial concentrations were highest in heavily populated workplaces, while the viable fungi concentrations were lowest in workplaces. The fungi and bacteria results showed high variation, which emphasises the importance of quality assurance (duplicates and field blanks) in the microbial field measurements. Our results indicate that personal exposure measurements of bioaerosols in indoor environments are feasible and supplement the information obtained by stationary samplers.  相似文献   

20.
气溶胶与降尘中多环芳烃的含量分布研究   总被引:13,自引:1,他引:12  
通过广东省茂名市区四个不同功能点大气气溶胶和降尘中多环芳烃的含量分布研究发现 :1、气溶胶中优控多环芳烃大大高于降尘中的含量 ,为降尘的 5.97~ 1 9.3倍 ;以石化厂区为例 ,非优控多环芳烃在气溶胶中的相对含量更高 ,为降尘的 2 4 .7倍。2、气溶胶中优控多环芳烃和非优控多环芳烃的分布为随分子量增加而含量增高的趋势 ,但降尘中优控多环芳烃的高含量相对集中于萤蒽至苯并 (b)萤蒽之间。3、不同功能区由于排放源的差别所表现出的气溶胶和降尘中优控多环芳烃总量及总量比值、部分强致癌和致癌物含量及含量比值均存在差异。4、对气溶胶和降尘中多环芳烃研究可以对降尘中非优控多环芳烃降解和溶解量进行估算。以石化厂区为例 ,降尘中非优控多环芳烃比原始含量已减少76%。  相似文献   

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