Comparison of two biological aerosol sampling methods

Two biological aerosol samplers, the Andersen two-stage microbial impactor and the May three-stage glass impinger, were examined to determine the benefits and effectiveness of the May sampler compared with the Andersen sampler, one of the most widely accepted samplers. Side-by-side samples were collected during simulated wastewater spray irrigation dispersion studies. Escherichia coli colony counts and air concentrations were statistically treated to determine the dependability of the May results with respect to the Andersen results. After data pairs containing potentially overloaded Andersen counts were eliminated, a linear regression of the remaining data was performed. It indicates that although the May sampler reports 82% of the Andersen sampler value, the correlation between the two samplers is good with an r2 value of 0.84. This comparison indicates that although there are differences between the two samplers, they do give comparable results and that when both are used in a sampling program, they tend to complement each other.     

Evaluation of three portable samplers for monitoring airborne fungi

Airborne fungi were monitored at five sample sites with the Burkard portable, the RCS Plus, and the SAS Super 90 air samplers; the Andersen 2-stage impactor was used for comparison. All samplers were calibrated before being used simultaneously to collect 100-liter samples at each site. The Andersen and Burkard samplers retrieved equivalent volumes of airborne fungi; the SAS Super 90 and RCS Plus measurements did not differ from each other but were significantly lower than those obtained with the Andersen or Burkard samplers. Total fungal counts correlated linearly with Cladosporium and Penicillium counts. Alternaria species, although present at all sites, did not correlate with total count or with amounts of any other fungal genera. Sampler and location significantly influenced fungal counts, but no interactions between samplers and locations were found.     

Comparative study of airborne pollen counts located in different areas of the city of Córdoba (south-western Spain)

Airborne pollen counts are mainly determined using a volumetric suction sampler based on the impact principle, that is, a Hirst-type spore trap. As a consequence of their volumetric nature, samplers detect pollen from a wide area, and therefore, a single sampler is frequently used to acquire information on airborne pollen counts for the whole city. The main goal of the present study was to compare airborne pollen counts at two sites located at opposite ends (south-west vs. north-east) of the southern Spanish city of Córdoba, to assess the advantages and disadvantages of using more than one sampler in the city. Also, a comparative study was carried out using two samplers at the same site, in order to confirm the efficiency of the samplers. Results revealed that data from one volumetric sampler—located within a city of medium size with uniform topography and vegetation conditions—are sufficient to establish monitoring of the main airborne pollen types, the pollen seasons involved and the timing of peak counts. For clinical studies, however, data on pollen counts in specific areas of the city may be of value, since pollen intensity may vary from one district to another, mainly in the case of ornamental plants with a local distribution inside the city. Comparison of data obtained by the two samplers running at the same site indicated that potential inter-site differences could not be attributed to differences in sampler efficiency.     

Evaluation of three portable samplers for monitoring airborne fungi.

Airborne fungi were monitored at five sample sites with the Burkard portable, the RCS Plus, and the SAS Super 90 air samplers; the Andersen 2-stage impactor was used for comparison. All samplers were calibrated before being used simultaneously to collect 100-liter samples at each site. The Andersen and Burkard samplers retrieved equivalent volumes of airborne fungi; the SAS Super 90 and RCS Plus measurements did not differ from each other but were significantly lower than those obtained with the Andersen or Burkard samplers. Total fungal counts correlated linearly with Cladosporium and Penicillium counts. Alternaria species, although present at all sites, did not correlate with total count or with amounts of any other fungal genera. Sampler and location significantly influenced fungal counts, but no interactions between samplers and locations were found.     

Monitoring airborne fungal spores in an experimental indoor environment to evaluate sampling methods and the effects of human activity on air sampling.

Aerobiological monitoring was conducted in an experimental room to aid in the development of standardized sampling protocols for airborne microorganisms in the indoor environment. The objectives of this research were to evaluate the relative efficiencies of selected sampling methods for the retrieval of airborne fungal spores and to determine the effect of human activity on air sampling. Dry aerosols containing known concentrations of Penicillium chrysogenum spores were generated, and air samples were taken by using Andersen six-stage, Surface Air System, Burkard, and depositional samplers. The Andersen and Burkard samplers retrieved the highest numbers of spores compared with the measurement standard, an aerodynamic particle sizer located inside the room. Data from paired samplers demonstrated that the Andersen sampler had the highest levels of sensitivity and repeatability. With a carpet as the source of P. chrysogenum spores, the effects of human activity (walking or vacuuming near the sampling site) on air sampling were also examined. Air samples were taken under undisturbed conditions and after human activity in the room. Human activity resulted in retrieval of significantly higher concentrations of airborne spores. Surface sampling of the carpet revealed moderate to heavy contamination despite relatively low airborne counts. Therefore, in certain situations, air sampling without concomitant surface sampling may not adequately reflect the level of microbial contamination in indoor environments.     

Comparison of methods for quantitative determinations of airborne bacteria and evaluation of total viable counts.

Three different methods of estimating airborne bacteria were compared. An Anderson sampler, a slit sampler, an impinger, and filter samplers with gelatine filters or membrane filters were tested for collection efficiency. The comparisons were made in laboratory experiments with an aerosol of Staphylococcus epidermidis or Serratia marcescens, in field experiments in two different industries, i.e., cotton mill and sewage plant, and in experiments with skin fragment sampling. Experiments were also performed estimating the total number of viable microorganisms on the airborne particles. The Andersen sampler gave the highest bacterial counts in all environments tested. The slit sampler gave statistically lower counts only in the aerosol experiments and cotton mill experiments, where the size of the majority of the particles carrying visible bacteria was 2 to 6 micrometers or smaller. In the sewage plant and skin fragment experiments, where the particles were mainly 5 micrometers or larger, the difference was not significant. The filters were efficient in sampling in skin fragment experiments only. With the agar impingement method, the total viable cell count showed a rising index value with increasing particle size. A mean of 13 bacteria was found per particle in the cotton mill, a mean of 24 in the sewage plant, and a mean of 147 in skin fragment experiments.     

Outdoor airborne fungi: sampling strategies

Summary A six-stage Andersens's sampler was compared with a single stage type (SAS) for the collection of airborne fungi particles. The efficiency of two non selective culture media, and namely malt agar (MA) and potato dextrose agar (PDA), which were both modified in order to inhibit bacterial growth, was compared as well. An overall assessment of quantitative results suggests that the two samplers have a comparable degree of efficiency. However, the SAS appears to collect a greater number of CFU/m³ in absence of wind and viceversa the Andersen one appears to be more efficient in the presence of wind. The number of species detected with the SAS device is smaller, thus requiring an increased number of suctions. Andersen sampler sieving, according to the aerodynamic characteristics of the particles, appears to be not too accurate: there is an overlap from stage to stage of the sampler and some larger particles settle on smaller particle-collecting stages and viceversa. The PDA, with streptomycin and chloramphenicol, is better and closer to the natural distribution pattern: the number of CFU/m³ is higher although the number of the collected species is about the same.     

Use of personal sporetraps to complement continuous aerobiological monitoring

We analysed pollen and spore data obtained from one continuous and two personal Burkard sporetraps during the spring months of three years (2007–2009). For the statistical analysis, the data was normalised with a log transformation, and then subjected to an ANOVA and a Pearson correlation analysis. The best time to use the personal samplers was determined from 15 years of continuous aerobiological monitoring pollen data to be between 11:00–16:00, when highest concentration was found and in a steady way. Height of sampling was compared at floor level and at 1.1 m with personal samplers; both of them were on a terrace at 6 m above the ground, but no statistically significant differences were found. The results revealed that there were apparently no differences between continuous and personal Burkard samplers for total pollen and spores. Nevertheless, distinguishing the main pollen types (i.e., Poaceae, Quercus, Olea, Cupressaceae, Plantago, and Platanus) revealed that there are some differences for Poaceae pollen only. In conclusion, personal samplers could be used to anticipate continuous monitoring data because their sampling is shorter and the results may be obtained quicker than with a continuous sampler, although they must never be considered as a replacement.     

Precision of the all-glass impinger and the andersen microbial impactor for air sampling in solid-waste handling facilities.

A method was devised to determine the precision of the all-glass impinger and the Andersen six-stage microbial impactor over a wide range of aerosol concentrations like those found in facilities which process solid waste. Simultaneous samples were collected inside a municipal solid-waste recovery system, and the data were treated statistically to estimate the precision of each air-sampling device. All-glass impingers yielded colony counts which indicated a linear relationship between samplers over an observed aerosol concentration of 1.1 X 10(3) to 2.8 X 10(7) colony-forming units per m3 of air. Impactors also yielded colony counts which indicated a linear relationship over an observed aerosol concentration range of 3.9 X 10(3) to 1.9 X 10(5) colony-forming units per m3 of air. The coefficients of variation for the all-glass impinger and the six-stage impactor in an environment with a high and variable dust level were determined to be 0.38 and 0.23, respectively.     

Dispersal of Pseudocercosporella herpotrichoides Spores from Infected Wheat Straw

Evidence from spore samples collected amongst infected straw spread on fallow ground supported the conclusion that spores of Pseudocercosporella herpotrichoides are dispersed mostly by rainsplash. Most spores travelled a short distance in the larger ballistic splash droplets, although some may have travelled further in smaller airborne droplets. Weekly spore counts from microscope slides under rainshields, a funnel and an impinger, evaluated as samplers for spores of P. herpotrichoides, showed a similar seasonal pattern. The funnel, as the largest sampler, generally collected most spores, but the impinger collected more spores per unit area of sampling surface. Slides sometimes collected spores when none was recovered from other samplers. Young wheat plants, exposed with the samplers and changed weekly, subsequently developed eyespot symptoms for most of the season.     

On the efficiency and correction of vertically oriented blunt bioaerosol samplers in moving air

The aspiration efficiency of vertical and wind-oriented Air-O-Cell samplers was investigated in a field study using the pollen of hazel, sweet chestnut and birch. Collected pollen numbers were compared to measurements of a Hirst-type Burkard spore trap. The discrepancy between pollen counts is substantial in the case of vertical orientation. The results indicate a strong influence of wind velocity and inlet orientation relative to the freestream on the aspiration efficiency. Various studies reported on inertial effects on aerosol motion as function of wind velocity. The measurements were compared to a physically based model for the limited case of vertical blunt samplers. Additionally, a simple linear model based on pollen counts and wind velocity was developed. Both correction models notably reduce the error of vertically oriented samplers, whereas only the physically based model can be used on independent datasets. The study also addressed the precision error of the instruments used, which was substantial for both sampler types.     

Airborne pollen grain concentrations at two different heights

The present study describes the airborne pollen grain concentrations at two different heights (1.5 m and 15 m, respectively). The survey was carried out in 1991 and 1992, using two Burkard spore-traps, both set up at the University of Córdoba, Faculty of Sciences. Generally, and for all herbaceous plants, pollen detection started and ended around the same date on both samplers. However, in the case ofOlea europaea, the pollen was detected in advance by the sampler located at 1.5 m compared with the one located at 15 m, probably due to the fact that olives growing close to the low sampler flower before the great olive plantations located some 60 km south of the city. No significant differences between the counts of both samplers have been observed, except in the case of Urticaceae, where the sampler situated on top of the building recorded higher pollen concentrations in both years. Similar annual peaks of Urticaceae are probably due to the buoyancy of their small, light grains and the explosive pollination mechanism which liberates pollen grains from the anthers of the Urticaceae family, includingUrtica andParietaria.     

Alfalfa weevil (Coleoptera: Curculionidae) larval sampling: comparison of shake-bucket and sweep-net methods and effect of training

Two field-sampling methods, shake-bucket and sweep-net, were compared for use in monitoring alfalfa weevil, Hypera postica (Gyllenhal), larvae in alfalfa hay of the high plains and intermountain region of North America. In this region, alfalfa grows to sufficient height to use both methods before the more damaging late instars peak in abundance. Both methods also were compared with extracting larvae by using Berlese funnels in the laboratory. The shake-bucket method was more sensitive in detecting small larvae (first and second instars) than large larvae (third and fourth instars), and the sweep-net method detected a lower proportion of small larvae. The number of larvae collected with the shake-bucket method was strongly correlated with number of larvae recovered from Berlese funnels (total larval counts, R2 = 0.85). Correlation of the sweep-net samples with the Berlese extraction was also significant but less strong (R2 = 0.56). In addition, sampler instruction was evaluated to determine whether demonstration training affects performance of inexperienced samplers using the two field-sampling methods. Training did not significantly change sampler performance in using the shake-bucket but did increase the number of large larvae when using the sweep-net. In addition, less variability was associated with the shake-bucket sampling method than with the sweep-net method for samplers who only had access to written sampling instructions. Therefore, when estimation of small larval abundance is desired for economic decision-making and sampling is performed by people with little or infrequent sampling experience, such as growers, the shake-bucket method is the preferred field-sampling method in the high plains and intermountain region of North America Sweep-net sampling is more variable than shake-bucket sampling, although demonstration training improves the usefulness of the sweep-net.     

A comparative study of three air-lift samplers used for sampling benthic macro-invertebrates in rivers

SUMMARY. From 16 air-lift samplers described in the literature, three were selected for operation from a small boat: Mackey, Pearson et al. and Verollet & Tachet samplers. Random samples (number of sampling units n= 10) were taken in a large tank with a known number of dried peas, representing invertebrates, amongst stones of uniform size. Separate experiments were performed with three sizes of stones (modal sizes 2–4, 16–20 and 32–36 mm). Stratified-random samples (usually n= 10) were taken in rivers at lour sites with modal particle sizes of < 0.1 2–4. 32–128 and 64–128 mm. The samplers were compared with a Ponar grab in the tank experiments and the field, and with a Naturalist's dredge in the field. Rates of pumping at different air-flows were established for each sampler. Water-How increased with increasing air-How and increasing submergence of the riser-pipe. The Mackey sampler achieved the highest water-flow and lifted more substratum and larger stones (64–128 mm) than the other samplers. The Verollet & Tachet sampler had the lowest pumping rate and failed to lift stones > 8 mm long, whilst the Pearson et al. sampler, with an intermediate performance, lifted mud and stones < 32 mm long. In the tank experiments, the area from which the Mackey and Pearson et al. samplers collected peas varied with air-flow, operating time and type of substratum. The Maekey sampler over-sampled and the Pearson et al. sampler usually under-sampled their respective sampling areas, except the latter sampler at high air-flows gave approximately quantitative results comparable to those of the Ponar grab for peas on the surface of stones 16–20 mm. No sampler collected much at a depth of 3 cm on larger stones (32–36 mm), and the Verollet & Tachet sampler failed completely on all substrata. In field trials, the Mackey and Pearson et al. samplers provided good qualitative samples except on large stones (32–128mm). The Verollet & Tachet sampler lifted little material. The relative abundance of taxa often differed between samplers. Estimates of the number of invertebrates per m² differed widely between samplers except for the Pearson et al. and Ponar grab which gave similar quantitative results at three sites. Values were very low for the Verollet & Tachet sampler and Naturalist's dredge and always very high for the Mackey sampler which often grossly over-sampled its sampling area. Therefore, these three samplers cannot be considered as quantitative. The relationship between the variances and the means of samples of peas in the tank and most invertebrates in the field followed a power law with values of the exponent b in the range 1.13–2.51. The present study completes the evaluation of 14 samplers used to sample benthic macro-invertebrates in deep rivers, and the more important conclusions from the comparative studies of seven grabs, four dredges and three air-lift samplers are summarized in a table.     

Bioaerosol mass spectrometry for rapid detection of individual airborne Mycobacterium tuberculosis H37Ra particles

Single-particle laser desorption/ionization time-of-flight mass spectrometry, in the form of bioaerosol mass spectrometry (BAMS), was evaluated as a rapid detector for individual airborne, micron-sized, Mycobacterium tuberculosis H37Ra particles, comprised of a single cell or a small number of clumped cells. The BAMS mass spectral signatures for aerosolized M. tuberculosis H37Ra particles were found to be distinct from M. smegmatis, Bacillus atrophaeus, and B. cereus particles, using a distinct biomarker. This is the first time a potentially unique biomarker was measured in M. tuberculosis H37Ra on a single-cell level. In addition, M. tuberculosis H37Ra and M. smegmatis were aerosolized into a bioaerosol chamber and were sampled and analyzed using BAMS, an aerodynamic particle sizer, a viable Anderson six-stage sampler, and filter cassette samplers that permitted direct counts of cells. In a background-free environment, BAMS was able to sample and detect M. tuberculosis H37Ra at airborne concentrations of >1 M. tuberculosis H37Ra-containing particles/liter of air in 20 min as determined by direct counts of filter cassette-sampled particles, and concentrations of >40 M. tuberculosis H37Ra CFU/liter of air in 1 min as determined by using viable Andersen six-stage samplers. This is a first step toward the development of a rapid, stand-alone airborne M. tuberculosis particle detector for the direct detection of M. tuberculosis bioaerosols generated by an infectious patient. Additional instrumental development is currently under way to make BAMS useful in realistic environmental and respiratory particle backgrounds expected in tuberculosis diagnostic scenarios.     

A microbial sampler for freshwaters

A freshwater sampler using five sterile evacuated glass tubes is described. Water enters when a rubber stopper is mechanically removed from the end of a sterile hypodermic needle inserted into each tube. Plate counts of bacterial colonies were compared with those obtained with other samplers.     

Viable fungi in corn dust

Numbers of viable fungal propagules in corn dusts in southern Georgia were estimated during various farm and grain elevator operations in 1979, 1980, and 1982. A six-stage Andersen sampler for viable microbial particles was used to sample the dusts with various agar media. The most abundant fungi in corn dusts were species of yeasts: Aspergillus, Penicillium, Cladosporium, Alternaria. Helminthosporium, and Fusarium. However, the relative abundance of these fungi differed between years. There was a greater incidence of the Aspergillus flavus group in the hot, dry year of 1980 compared with the cooler, wetter years of 1979 and 1982. Fungi in the corn dusts sampled numbered between 10(4) and 10(9) viable propagules per m3 of air. By contrast, outdoor air often contained fewer than 10(4) viable fungal propagules per m3. Most A. flavus propagules were deposited at stages three and four of the Andersen sampler, with correspond to the trachea, primary bronchi, and secondary bronchi in the human respiratory system. In an assessment of the air spores by exposing sterile petri dishes, more large-spored fungi, like Alternaria tenuis, and fewer small-spored fungi, such as A. flavus, were detected when compared with colony counts from petri dishes exposed to air in the Anderson sampler.     

Viable fungi in corn dust.

Numbers of viable fungal propagules in corn dusts in southern Georgia were estimated during various farm and grain elevator operations in 1979, 1980, and 1982. A six-stage Andersen sampler for viable microbial particles was used to sample the dusts with various agar media. The most abundant fungi in corn dusts were species of yeasts: Aspergillus, Penicillium, Cladosporium, Alternaria. Helminthosporium, and Fusarium. However, the relative abundance of these fungi differed between years. There was a greater incidence of the Aspergillus flavus group in the hot, dry year of 1980 compared with the cooler, wetter years of 1979 and 1982. Fungi in the corn dusts sampled numbered between 10(4) and 10(9) viable propagules per m3 of air. By contrast, outdoor air often contained fewer than 10(4) viable fungal propagules per m3. Most A. flavus propagules were deposited at stages three and four of the Andersen sampler, with correspond to the trachea, primary bronchi, and secondary bronchi in the human respiratory system. In an assessment of the air spores by exposing sterile petri dishes, more large-spored fungi, like Alternaria tenuis, and fewer small-spored fungi, such as A. flavus, were detected when compared with colony counts from petri dishes exposed to air in the Anderson sampler.     

Evaluation of Four Aerobiological Sampling Methods for the Retrieval of Aerosolized Pseudomonas syringae

The Andersen six-stage impactor, the SAS (Surface Air System) impactor, the AGI-30 impinger, and gravity plates were evaluated for the retrieval of aerosol-released Pseudomonas syringae. The upper limits of the impactor samplers were exceeded at a spray concentration of 10⁷ CFU/ml, indicating that these samplers are not appropriate for monitoring high airborne concentrations. Decreased cell concentrations were retrieved with increased sampling time for the Andersen and AGI samplers, indicating that a minimum sampling time is preferable for monitoring aerosolized vegetative cells.     

Direct Method for Determining the Viability of a Freshly Generated Mixed Bacterial Aerosol

A direct method was developed to determine the viability of a freshly generated mixed bacterial aerosol. A mixed suspension of (32)P-labeled Staphylococcus aureus and (35)S-labeled Proteus mirabilis was nebulized, and the aerosol was collected and separated according to particle size with an Andersen sampler. Quantitative and qualitative bacteriological and radioisotopic techniques were used to obtain ratios of bacterial to radioactive counts for each organism in samples of the nebulizer suspension and aerosol. Loss of viability was calculated from the change that occurred between the ratio of the nebulizer suspension and the ratio of the aerosol. The viability of S. aureus was unaffected by aerosolization, whereas the viability of P. mirabilis declined by 20 to 60% and was inversely proportional to particle size. The advantages of this method over present indirect methods, as well as potential applications of the method, are discussed.