Bees Scavenge Airborne Bacteria.

An air conditioned wind tunnel system was designed, fabricated, and tested to determine whether tethered bees scavenge microbeads or Bacillus subtilis var. niger spores from aerosols. Tests showed that microbeads and spores were scavenged by bumblebees and honeybees, respectively. Five independent variables and their interactions were used in a stepwise multiple regression. Two of them, the cube root of the electrostatic charge on the honeybee and the dose of the spore aerosol, accounted for most of the statistically significant fit to the model's two dependent variables: the percentage of the dose adsorbed by honeybees and the number of spores adsorbed by the same bees. Both dependent variables increased directly so that an increase in electrostatic charge on the bee (i.e., cube root 32 pC) resulted in an increase (i.e., approximately 1%) in the spore dose adsorbed and the number of spores adsorbed by the bees. It was theorized that the spores were in an adsorption/desorption equilibrium that responded to the concentration "pressure" of the spore aerosol. Further, the charge on the bee affected the adsorption force on the bee's surface, as well as increasing the effective aerosol volume accessible for the bee's scavenging. In short, relating these findings to bees scavenging bacteria from the ambient atmosphere, it appears that the spore exposure (where exposure means the product of the ambient concentration, the time the bee is exposed, and air volume through which the bee flies) controls the number of spores adsorbed by a bee, and the static charge on the bee controls the adsorption/desorption equilibrium and presumably the scavenging volume.

Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying.

The microorganisms Escherichia coli DH5 alpha and Bacillus thuringiensis HD-1 show an increased tolerance to freeze-drying when dried in the presence of the disaccharides trehalose and sucrose. When the bacteria were dried with 100 mM trehalose, 70% of the E. coli and 57% of the B. thuringiensis organisms survived, compared with 56 and 44%, respectively, when they were dried with sucrose. Only 8% of the E. coli and 14% of the B. thuringiensis organisms survived drying without the sugars. Fourier transform infrared spectroscopy was used to investigate the role of membrane phase transitions in the survival of the organisms during drying and rehydration. Both E. coli and B. thuringiensis showed an increase of 30 to 40 degrees C in the temperature of their phospholipid phase transition when dried without the sugars, while phase transition temperatures of those dried with the sugars remained near those of the hydrated cells. A Fourier transform infrared spectroscopy microscope made it possible to investigate the effects of drying on the protein structure in the intact cells. The amide II peak shifts from 1,543 cm-1 in the hydrated cells to about 1,533 cm-1 in the cells dried without sugar. There is no shift in the amide II peak when the cells are dried with trehalose or sucrose. We attribute the increased survival to the sugars' ability to lower the membrane phase transition temperature and to protect protein structure in the dry state.(ABSTRACT TRUNCATED AT 250 WORDS)

Airborne Bacteria in the Atmospheric Surface Layer: Temporal Distribution above a Grass Seed Field.

Temporal airborne bacterial concentrations and meteorological conditions were measured above a grass seed field in the Willamette River Valley, near Corvallis, Oreg., in the summer of 1993. The concentration of airborne bacteria had a maximum of 1,368.5 CFU/m(sup3), with a coefficient of variation of 90.5% and a mean of 121.3 CFU/m(sup3). The lowest concentration of bacteria occurred during the predawn hours, with an average of 32.2 CFU/m(sup3), while sunrise and early evening hours had the highest averages (164.7 and 158.1 CFU/m(sup3), respectively). The concentrations of bacteria in the atmosphere varied greatly, with a maximum difference between two 2-min samples of 1,995 CFU/m(sup3). The concentrations of bacteria in the atmosphere could be divided into five time periods during the day that were thought to be related to the local diurnal sea breeze and Pacific Coast monsoon weather conditions as follows: (i) the nighttime minimum concentration, i.e., 2300 to 0600 h; (ii) the sunrise peak concentration, i.e., 0600 to 0800 h; (iii) the midday accumulating concentration, i.e., 0800 to 1515 h; (iv) the late-afternoon sea breeze trough concentration, i.e., 1515 to 1700 h; and (v) the evening decrease to the nighttime minimum concentration, i.e., 1700 to 2300 h. The sunrise peak concentration (period ii) is thought to be a relatively general phenomenon dependent on ground heating by the sun, while the afternoon trough concentration is thought to be a relatively local phenomenon dependent on the afternoon sea breeze. Meteorological conditions are thought to be an important regulating influence on airborne bacterial concentrations in the outdoor atmosphere in the Willamette River Valley.

Protection of freeze-dried Escherichia coli by trehalose upon exposure to environmental conditions.

Freeze-dried cultures of wild-type and genetically engineered strains of Escherichia coli lost their colony-forming ability upon exposure to air, visible light, and certain relative humidity levels. Both strains could be maximally protected from these lethal effects with 100 mM trehalose, a concentration calculated to just saturate the interphospholipid spaces in the cell membrane, thus preserving the liquid-crystalline structure. The trehalose protection was observed for at least 96 h. Trehalose increased viability as much as 2000-4000% over nontreated populations. In all cases, exposure to environmental conditions was more damaging to the genetically engineered strain.

Survival differences among freeze-dried genetically engineered and wild-type bacteria.

Because the death mechanisms of freeze-dried and air-dried bacteria are thought to be similar, freeze-drying was used to investigate the survival differences between potentially airborne genetically engineered microorganisms and their wild types. To this end, engineered strains of Escherichia coli and Pseudomonas syringae were freeze-dried and exposed to air, visible light, or both. The death rates of all engineered strains were significantly higher than those of their parental strains. Light and air exposure were found to increase the death rates of all strains. Application of death rate models to freeze-dried engineered bacteria to be released into the environment is discussed.

Resuscitation effects of catalase on airborne bacteria.

Catalase incorporation into enumeration media caused a significant increase (greater than 63%) in the colony-forming abilities of airborne bacteria. Incubation for 30 to 60 min of airborne bacteria in collection fluid containing catalase caused a greater than 95% increase in colony-forming ability. However, catalase did not have any effects on enumeration at high relative humidities (80 to 90%).

Trajectory of aerosol droplets from a sprayed bacterial suspension.

Simulated droplet trajectories of a polydispersed microbial aerosol in a laminar air flow regimen were compared with observed dispersal patterns of aerosolized Bacillus subtilis subsp. niger spores in quasilaminar airflow. Simulated dispersal patterns could be explained in terms of initial droplet sizes and whether the droplets evaporated to residual aeroplanktonic size before settling to the ground. For droplets that evaporated prior to settling out, a vertical downwind size fractionation is predicted in which the microbial residue of the smallest droplets settles the least, and is found in the airstream at about sprayer height, and progressively larger droplet residues settle to progressively lower heights. Observations of spore particle size distributions downwind from a spray source support the simulation. Droplet and particle size distributions near the source had three size fractions: one containing large, presumably nonevaporated droplets of greater than or equal to 7 microns in diameter, and two smaller fractions, with diameters of 2 to 3 microns (probably the residue of droplets containing more than one spore) and 1 to 2 microns (probably the residue from single-spore droplets). As predicted by the simulation, the aerosol settled and progressed downwind, with the number of small droplets and particles increasing in proportion to the height and distance downwind.

Effects of betaine on enumeration of airborne bacteria.

The osmoprotectant betaine was incorporated into collection fluid and enumeration medium to determine its effects on the colony-forming abilities of airborne bacteria, which were collected from three separate locations: a wastewater treatment plant, the roof of a laboratory building, and an unobstructed farmland. At all locations, addition of 2 to 5 mM betaine caused a significant increase (from 21.6 to 61.3%) in colonial outgrowth, compared with the growth rate of controls without betaine. The presence of betaine in both the collection fluid and the enumeration medium had an additive effect on the colony-forming ability of airborne bacteria compared with the presence of betaine in either one alone. The effect of various betaine concentrations on the enumeration of aerosolized Pseudomonas syringae was determined. Betaine showed a threshold for maximum effect at a concentration of 2 to 5 mM. At higher concentrations (10 to 20 mM), the effects of betaine were negligible or possibly inhibitory. The significance of these results with respect to the development of protocols for monitoring airborne microorganisms, including genetically engineered microorganisms, is discussed.

Simulation of airborne microbial droplet transport.

The framework for a simulation model which describes the dispersion of individual droplets of water containing viable microbes is presented. The model accounts for physical, chemical, biological, and measured meteorological parameters of each droplet at each of many short time steps. Repeating the modeling process for many droplets will simulate a cloud of droplets. The model is compared with the Tulelake, Calif., release in 1988 and found to show very similar patterns of deposition within 30 m (the maximum observation distance of the source. A hypothesis for the survival sequence in the microbe-containing droplets is discussed.

Some Changes in Gut Bacterial Flora of Field-Grown Peridroma saucia (Lepidoptera: Noctuidae) When Brought into the Laboratory.

Removal of Peridroma saucia from the field to the laboratory caused little change in the quantity of facultative and aerobic bacteria in the gut but produced significant qualitative and quantitative changes in distinguishable groups of the family Enterobacteriaceae in the gut.

Estimating downwind concentrations of viable airborne microorganisms in dynamic atmospheric conditions.

A Gaussian plume model has been modified to include an airborne microbial survival term that is a best-fit function of laboratory experimental data of weather variables. The model has been included in an algorithm using microbial source strength and local hourly mean weather data to drive the model through a summer- and winter-day cycle. For illustrative purposes, a composite airborne "virus" (developed using actual characteristics from two viruses) was used to show how wind speed could have a major modulating effect on near-source viable concentrations. For example, at high wind speeds such as those occurring during the day, or with short travel times, near-source locations experience high viable concentrations because the microorganisms have not had time to become inactivated. As the travel time increases, because of slow wind speed or longer distances, die-off modulation by sunshine, relative humidity, temperature, etc., potentially becomes increasingly predominant.

Microbial aerosols: estimated contribution of combine harvesting to an airshed.

From plate counts of the airborne microorganisms in the downwind dust plume of operating grass-seed combines, the mean source concentrations were calculated to be 6.4 x 10 and 4.7 x 10/m, respectively, potentially accounting for at least 41.9% of the bacteria and 35.1% of the fungi in the airshed in the Willamette Valley, Oregon.

Effects of certain cadmium species on pure and litter populations of microorganisms.

Cadmium inhibition of microorganisms was found to be bacterial and chemical species dependent. E. coli inhibition was a function of the cadmium-ion concentration irregardless of the presence of citrate, a chelator for cadmium that it could not metabolize. Whereas with a Pseudomonas sp. able to metabolize citrate, cadmium inhibition was a function of both the cadmium ion and the presence of citrate. With no citrate, inhibition of this organism occurred only at relatively high cadmium-ion concentrations (above 10(-4) M); when citrate was added to the same cadmium-containing growth medium, inhibition was observed at a 1000 times lower cadmium-ion concentration (i.e., 10(-7) M). This observation is contrary to the classical understanding where a chelate reduces the toxic form of a metal allowing increased growth of the organism. The species of cadmium also differentially inhibited the Douglas fir letter respiration and nitrogen-fixing community activities.

Enrichment of cadmium-mediated antibiotic-resistant bacteria in a Douglas-fir (Pseudotsuga menziesii) litter microcosm.

A set of Douglas-fir needle litter microcosms was amended with cadmium, acid, a combination of both, or neither. After 2 weeks of incubation, bacterial colony counts were made of litter homogenates inoculated onto agar media containing an antibiotic (streptomycin, chloromycetin, ampicillin, or gentamicin), cadmium, both, or neither. In all microcosms bacterial abundance was similar but the quality was very dissimiliar. Cadmium-treated microcosms had populations enriched for cadmium and gentamicin resistance and streptomycin and chloramphenicol sensitivity. Acid amendment had no consistent effect on the microcosm populations except that which could be attributed to the cadmium treatment amendment alone.

Estimation of downwind viable airborne microbes from a wet cooling tower-Including settling.

In recent years, reuse of municipal waste water as the coolant in drift-producing cooling towers at electrical generating plants has become increasingly common. A hueristic model is presented that can be used to estimate the concentrations of viable airborne microbes in the drift from a wet cooling tower given the concentration of microbes in the cooling tower. The purpose of this presentation is to allow the nonmeteorologist to understand the factors affecting airborne concentration and to make crude estimates of ground-level concentrations of airborne microorganisms. Concentrations are calculated using a standard meterological method, the Gaussian dispersion model, in which terms have been included for droplet settling and microbial death rate.

Estimation of viable airborne microbes downwind from a point source.

Modification of the Pasquill atmospheric diffusion equations for estimating viable microbial airborne cell concentrations downwind form a continuous point source is presented. A graphical method is given to estimate the ground level cell concentration given (i) microbial death rate, (ii) mean wind speed, (iii) atmospheric stability class, (iv) downwind sample distance from the source, and (v) source height.

A cluster analysis of some bacteria in the water column of Green Lake, Washington.

Mass inoculation and computer clustering techniques were used in an abbreviated procedure to group similar bacteria isolated from four depths in the water column of Green Lake, Washington. Four groups of bacteria were differentiable and were catagorized as orange-yellow Flavobacterium-Cytophaga, yellow Flavobacterium-Cytophaga, Vibrio-Aeromonas-, and Pseudomonas-like. Some of the groups were most prevalent in certain sample depths in the water column.

Survival of airborne bacteria in a high urban concentration of carbon monoxide.

Vegetative cells of Serratia marcescens 8UK, Sarcina lutea, and spores of Bacillus subtilus var. niger were held in aerosols, with and without an urban concentration of CO (85 muliters per liter or ppm), for up to 6 hr at 15 C and a relative humidity (RH) of approximately 0, 25, 50, 75, and 95%. It was found that CO enhanced the death rate of S. marcescens 8UK at least four- to sevenfold at low RH (ca. 1 to 25%), but protected the cells at high RH (ca. 90%). Death rates of S. lutea, with or without added CO, were comparatively low over the entire RH range. However, in the first hour, airborne S. lutea held in CO-containing air were more stable than those in air without added CO (i.e., CO protection). A marked increase in the death rate (up to 70-fold) occurred in the subsequent 5 hr within the RH range of approximately 0 to 75%. Statistical analysis indicated that aerosol decay rates of B. subtilus var. niger spores decreased significantly, when held in a CO-containing as compared to a non-CO-containing atmosphere, in the 0 to 85% RH range. Thus, the data presented indicate that CO in the urban environment may have a protective or lethal effect on airborne bacteria, dependent upon at least the microbial species, aerosol age, and relative humidity. A mechanism for CO death enhancement and protection of airborne S. marcescens 8UK is suggested to involve CO uncoupling of an energy-requiring death mechanism and an energy-requiring maintenance mechanism at high and low RH, respectively.