AN AUTOMATIC VOLUMETRIC SPORE TRAP

A suction trap has been made in which the spores entering a narrow orifice, directed into the wind, are impacted on a Vaseline-coated microscope slide moved across the orifice at 2 mm./hr. Estimates of spore content of the air can be made, with higher efficiency than by previous traps, at different times of day and thus be more closely correlated with variations in weather.
Wind-tunnel tests with spores of Lycopodium clavatum showed maximal and minimal efficiencies of 93.8 and 62.4% respectively, with a suction rate of 10.0 1./min., in the range of wind speeds from 1.5 to 9.3 m./sec.     

Dispersal of Septoria nodorum Pycnidiospores by Simulated Rain and Wind

The influence of wind on the splash dispersal of Septoria nodorum pycnidiospores was studied in a raintower/wind tunnel complex with single drops or simulated rain falling on spore suspensions or infected stubble with windspeeds of 1.5 to 4 m/sec. When single drops fell on spore suspensions (depth 0.5 mm, concentration 7.8 × 10⁵ spores/ml) most of the spore-carrying droplets collected on fixed photographic film between 0–4 m downwind (windspeed 3 m/sec) were >200 μm in diameter. However, most spores were carried in droplets with diameter > 1000 μm, 70 % of which carried more than 100 spores. When simulated rain fell on infected stubble most of the spore-carrying droplets collected beyond 1 m downwind (windspeeds 1.4 and 4 m/sec) were <200 μm in diameter and none were >600 μm; most of these droplets carried only one spore. The distribution of splash droplets (with diameter >100 μm) deposited on chromatography paper showed a maximum at 40–50 cm upwind of the target but many more droplets were deposited 20–30 cm downwind, when single drops fell on a spore suspension (concentration 1.2 × 10⁵ spores/ ml) containing fluorescein dye with a windspeed of 2 m/sec; droplets were collected up to 3 m downwind but not more than 70 cm upwind. With a windspeed of 3 m/sec, numbers of sporecarrying droplets and spores collected on film decreased with increasing distance downwind; most were collected within 2 m of the target but some were found up to 4 m. When simulated rain fell on infected stubble, increasing the windspeed from 1.5 to 4 m/sec greatly increased the number of spores deposited more than 1 m downwind. At 1.5 m/sec none were collected beyond 2 m downwind, whereas at 4 m/sec some were collected at 4 m. A few air-borne S. nodorum spores were collected by suction samplers at a height of 40 cm at distances up to 10 m downwind of a target spore suspension on which simulated rain fell.     

The Effect of Wind Speed on Assimilation Rate--a Re-assessment

Deneke concluded from wind-tunnel experiments that CO₂ uptakeis increased by wind, but that this stimulation is proportionatelyless at higher wind speeds, and that above Ioo m./min. (I₆₇cm./sec.) no further increase results. His data in fact indicatea linear relationship between wind and assimilation rate overthe whole range investigated. The wind speeds he used were ofthe same order as those occurring in vegetation in the field. Published experiments on the effect of wind on dry-weight increase,on the other hand, show that wind reduces growth; but theseexperiments have used winds equivalent to 'gale' or 'hurricane',and at such speeds other, harmful effects of wind become important. Wind-tunnel investigations on stands of Brassica napus reconcilethese observations by showing growth-rates to increase withwind speed up to an optimum near 30 cm./sec., and to fall offat higher speeds. It is considered that the stimulating effectof wind on assimilation shown by Deneke's and these experimentsis likely to be significant in the field.     

An Optimum Wind Speed for Plant Growth

Observations on the growth-rate of stands of young plants ofBrassica napus in a wind tunnel show that relative growth-raterises with wind speed at low wind speeds but falls again whenwind speed is further increased. A wind speed of 0.3m./sec.caused optimal growth in these experiments. Changes in relativegrowth-rate were small.     

Dispersal of Erysiphe graminis and Lycopodium clavatum spores near to the source in a barley crop

In experiments to study dispersal of spores in a crop of barley, a 4-m wide strip of the cultivar Zephyr (a mildew-susceptible variety) was a source of mildew (Eyrsiphe graminis) conidia. Small suction traps, previously calibrated in a wind tunnel, were used to measure spore concentration within and above the crop. Large concentrations of conidia were measured in the crop at the downwind edge of the Zephyr strip but these decreased rapidly with distance downwind. At 1 m concentrations were halved and by 4 m they were no greater than the background concentration measured in the crop upwind of the source. Next to the source, concentrations were much greater within than above the crop and net spore movement (flux) was upwards out of the crop; by 4 m downwind concentrations were greater above the crop and spore flux was reversed. Lycopodium clavatum spores were released in the same crop from a line of point sources. Concentrations also decreased rapidly downwind but, with no background of spores, numbers remained greater within than above the crop further from the source than for E. graminis. Even so by 7 m downwind concentrations in the crop had declined to less than those above. Deposition of L. clavatum spores onto horizontal glass slides in the crop agreed with that expected by settling. However, impaction onto vertical cylinders among plants was much greater than predicted. The reason is not known although turbulent air-flow around plants may, in some way, enhance impaction. Many E. graminis conidia near the source were deposited in clumps. This prevented any accurate prediction of deposition rates as fall speeds of clumps (necessary for prediction) were not known. Not surprisingly, deposition on horizontal slides often exceeded that expected from settling of single spores although it was not always greatest where clumps predominated. The proportion of spores deposited on vertical cylinders and horizontal slides located among plants ranged from 0–02 to 0–27 and from 0–019 to 0–127 of the area dose, respectively. Although these may seem to be small trapping efficiencies, the same deposition rates in a crop with many leaves and stems would rapidly filter most spores from air in the crop and can explain why concentrations were observed to decline so rapidly.     

Effects of meteorological conditions on spore plumes

Fungal spores are an ever-present component of the atmosphere, and have long been known to trigger asthma and hay fever symptoms in sensitive individuals. The atmosphere around Tulsa has been monitored for airborne spores and pollen with Burkard spore traps at several sampling stations. This study involved the examination of the hourly spore concentrations on days that had average daily concentrations near 50,000 spores/m(3) or greater. Hourly concentrations of Cladosporium, Alternaria, Epicoccum, Curvularia, Pithomyces, Drechslera, smut spores, ascospores, basidiospores, other, and total spores were determined on 4 days at three sites and then correlated with hourly meteorological data including temperature, rainfall, wind speed, dew point, air pressure, and wind direction. On each of these days there was a spore plume, a phenomenon in which spore concentrations increased dramatically over a very short period of time. Spore plumes generally occurred near midday, and concentrations were seen to increase from lows around 20,000 total spores/m(3) to highs over 170,000 total spores/m(3) in 2 h. Multiple regression analysis of the data indicated that increases in temperature, dew point, and air pressure correlated with the increase in spore concentrations, but no single weather variable predicted the appearance of a spore plume. The proper combination of changes in these meteorological parameters that result in a spore plume may be due to the changing weather conditions associated with thunderstorms, as on 3 of the 4 days when spore plumes occurred there were thunderstorms later that evening. The occurrence of spore plumes may have clinical significance, because other studies have shown that sensitization to certain spore types can occur during exposure to high spore concentrations.     

Ventilation required to entrain small particles from leaves

Particles are blown from leaves when the wind at the height of the particles exceeds a minimum which is about 5 m/sec for some fungal spores. In the moderate winds typical within a canopy of leaves, the minimum is attained at spore height during brief changes in wind or puffs before the boundary layer grows to particle height. The requisite change in speed to remove spores occurs over a sizeable area only when the speed changes abruptly in a short distance in the direction of the wind.     

The Effect of Artificial Wind on the Growth-rate of Plants in Water Culture: With Plate and three Figures in the Text

Young plants of Brassica napus (rape), Hordeum vulgare, andPisum sativum growing in water culture have been exposed tofour continuous wind speeds for from 4 to 5 days. The plantswere exposed to natural daylight, humidity, and temperaturein a wind tunnel in which only air movement was controlled.The wind speeds found among the plants in the four sectionsof this tunnel were approximately 0·3, 0·7, 1·7,and 4·0 m./sec. The results showed no significant changein relative growth-rate or net assinilation rate with wind speed. Previously published results, obtained with plants in soil orsand culture, have differed from those in the present experimentsin showing a fall in the amount of growth as wind speed increased.It is suggested that the reduction in growth found in theseprevious experiments was caused by partial drying out of theplants, whilst in the present experiments water-supply to theroots was abundant and this effect was considerably reduced.     

Sequential Use of Wright's and Ziehl-Neelsen's Stains for Demonstrating Phagocytosis of Bacterial Spores

Nongerminating spores, germinating spores, and vegetative cells of Clostridium botulinum type A were observed during phagocytosis in the peritoneal fluid of white mice. Since phagocytes are easily ruptured by heat, the method described avoids heating, as this has been employed in conventional spore staining methods. A thin smear of the fluid is air dried on the slide for 2 hr, and stained by Wright's method: stain, 2 min; dilution water, 2 min; and rinsed; then in 0.005% methylene blue for 30 sec, and rinsed. This is followed by Ziehl-Neelsen's stain for 3-4 min and destained with 1: acetone-95% ethanol for 10 sec. The slide is rinsed, and Wright's staining repeated: stain 1 min, dilution 2-3 min; and thereafter washed about 5 ml of Wright's buffer. Blotting and air drying completes the staining. Non-germinating spores stain light red with a red spore wall, germinating spores are deep red throughout, vegetative cells are blue, and leucocytes show a dark purple nucleus and light blue cytoplasm.     

Solving the aerodynamics of fungal flight: how air viscosity slows spore motion

Viscous drag causes the rapid deceleration of fungal spores after high-speed launches and limits discharge distance. Stokes' law posits a linear relationship between drag force and velocity. It provides an excellent fit to experimental measurements of the terminal velocity of free-falling spores and other instances of low Reynolds number motion (Re<1). More complex, non-linear drag models have been devised for movements characterized by higher Re, but their effectiveness for modeling the launch of fast-moving fungal spores has not been tested. In this paper, we use data on spore discharge processes obtained from ultra-high-speed video recordings to evaluate the effects of air viscosity predicted by Stokes' law and a commonly used non-linear drag model. We find that discharge distances predicted from launch speeds by Stokes' model provide a much better match to measured distances than estimates from the more complex drag model. Stokes' model works better over a wide range projectile sizes, launch speeds, and discharge distances, from microscopic mushroom ballistospores discharged at <1 m s(-1) over a distance of <0.1mm (Re<1.0), to macroscopic sporangia of Pilobolus that are launched at >10 m s(-1) and travel as far as 2.5m (Re>100).     

Settling rates and density of spores of Henneguya doori (Myxosporea) in water.

Using Henneguya doori from Perca flavescens (yellow perch), the current study provides a method for examining settling rates, attitudes, and densities of myxosporean spores. Fresh and fixed spores settled similarly at 8.34 x 10(-6) m/sec and 8.53 x 10(-6) m/sec, respectively. The spores settled in the anterior end first, did not rotate, and had a specific gravity of 1.062. Comparison of settling rates proved more effective for determining spore density than direct chemical estimation. The slow rate of settling permits effective dispersal in open water.     

Microbial validation of vent filters

The Upjohn Company uses filtration to remove microorganisms and particulates from air and other gases which may come in contact with sterile products. To validate the microbial retentivity of these filters, they were challenged with an aerosol of Bacillus subtilis var niger spores. An aerosol challenge was used because it more closely simulated the use for which these filters were designed. The test apparatus was constructed of autoclavable components using a jet-type nebulizer and heated air mixing tube. Characterization of the aerosol particle size distribution with a particle size analyzer demonstrated that 80% of the particles had a diameter of x 3.0 times;m and that the particles had a mean mass diameter of 1.9 times;m with a geometric standard deviation of 1.8 times;m. Studies conducted with aerosols of Bacillus subtilis var niger spores demonstrated that the test apparatus could recover ca. 50% of the spores that were aerosolized. Hydrophobic filters from various manufactures were challenged with an aerosol of at least 10(8) spores of Bacillus subtilis. All filters tested could retain at least 10(9) spores when physical integrity of the filter was verfield.     

Spread of directly evoked responses in the cat''s cerebral cortex

A study has been made of the electrical responses to direct stimulation of the exposed cerebral cortex of cats that had been immobilized with neuromuscular blocking drugs, and whose muscle and skin wounds had been locally anesthetized. The characteristics and spread of the first and second surface-negative responses are described. It was found that the first surface-negative response to weak stimuli decays linearly to zero at 3 to 6 mm. from the point of stimulation. Intermediate stimuli cause farther and non-linear spread: responses are re-initiated, or reinforced, at 6 to 10 mm.; and supramaximal stimulation produces reinforcement both at 5 and at 10 mm. The conduction velocity of these responses is uniform for linear spread (0.7 to 2.0 m./sec.), but reinforced responses occur 1 to 3 msec. earlier than would be expected for simple conduction. The phenomenon of re-initiation, or reinforcement, depends upon the excitatory state of the brain; circulation and previous stimulation are important factors. Connections outside the gyrus matter only in so far as they provide other sources of general excitation. It is concluded that two types of transmission: slow and fast, can lead to generation of similar surface-negative responses. The suggestion is made that the slowly conducted surface-negative potentials are due to direct or to synaptic excitation of pyramidal cells; while the responses with shortened latency are initiated synaptically on other pyramidal cells after fast conduction at about 10 m./sec. in tangential fibres.     

Observations on the production and dispersal of spores, and infection by Rhynchosporium secalis

The numbers of spores of Rhynchosporium secalis washed from samples of barley plants taken weekly, varied markedly. No consistent association with amount of previous rainfall or length of the preceding dry period was detected. On potted seedlings exposed within a crop most infection occurred during long rain periods or when rain fell late in the evening; fewest lesions usually developed on seedlings prevented from touching adjacent plants. Rotorod traps fitted with a 13 mm diameter disc at the apex of each arm were operated under 24 cm diameter covers. Spores were collected on circular cover slips fixed to each disc with glycerine jelly. At all stages of crop growth more spores were trapped at ground level than at other heights tested up to 1 m. The number of spores trapped was not related to the quantity of or duration of rainfall but was related to the mean rate of fall during brief showers only. Efficiency of droplet retention was assessed in a wind tunnel. It declined rapidly when more than c. 5 pl was presented to the disc surface and was less at a wind speed of 1.0 than 0.5 m s-^l. Spore distribution on discs indicated that spores were washed from the surface during long rain periods.     

Spore Dispersal and Plant Disease Gradients; a Comparison between two Empirical Models

Power law and exponential models were fitted to 325 sets of observations which described decreases with distance in deposition of air-borne or splash-borne spores, or pollen, or in amounts of plant disease caused by fungi, bacteria or viruses. There, was generally little difference between the models in the goodness of fit to these data, although deposition gradients for spores borne in splash droplets were fitted better by exponential models and gradients for fungi with air-borne spores less than 10 μm in diameter were fitted better by power law models. The exponential model has the property that the observed variable decreases by half as the distance from the source increases by a constant increment (the half-distance); this provides a measureof the gradient that is more easy to visualize than the exponent in power law model. The half-distances of gradients for air-borne pathogens were greater than those for splash-borne or soil-borne pathogens. The exponential model is easier to incorporate into models of disease development than the power law model because the boundary condition at the source (the estimated number of spores or amount of disease at the source) is finite rather than infinite. However, both these empirical models have limitations and should not be extrapolated to distances outside the observed range.     

THE EFFECT OF NISIN ON THE GROWTH OF CELLS AND SPORES OF CLOSTRIDIUM WELCHII IN GELATINE

SUMMARY: The action of various concentrations of nisin on the development of cells and spores from untreated and heated suspensions of Clostridium welchii added to gelatine has been investigated, using a tube colony count technique. The development of vegetative cells was prevented by the presence of 2 Reading Units (R.U.) of nisin/ml of the final culture medium, although the effective concentration may have been 6 R.U./ml. Approximately 40 R.U./ml prevented colony formation from spores. Gelatine containing nisin was dried. A considerable proportion of the activity of the antibiotic was still present after storage for 5 weeks.     

Deposition of spores and other particles on vegetation and soil

The rate of deposition of 20–30 μm diameter particles, including spores and pollen grains, on plant and other surfaces, is determined, first, by the frequency at which particles strike the surfaces and, secondly, by the proportion retained on the surface rather than rebounding into the airstream. Spores and pollen grains tagged with a radioactive marker were used to show that the impaction efficiency on leaves and stems depends very much on whether or not the surfaces are sticky or moist. If they are, the rate of deposition may approach that predicted aerodynamically. If the plant surfaces are dry, there is saltation of some spores and the effective rate of deposition is greatly reduced.     

The relative sensitivity of Renshaw cells to orthodromic group Ia volleys caused by static stretch and vibrations of extensor muscles.

1. Activity of Renshaw cells monosynaptically excited by ventral root stimulation and disynaptically excited by electric stimulation of the group Ia afferents in the gastrocnemius-soleus (GS) nerve, was recorded in precollicular decerebrate cats. The response of these units to prolonged vibration applied longitudinally to the deefferented GS muscle was then compared with that elicited by static stretch of the homonymous muscle, for comparable frequencies of discharge of the group Ia afferents. 2. Small-amplitude vibration of the GS muscle at 200/sec for one second produced a sudden increase in the discharge rate of Renshaw cells, which gradually decreased within the first 100 msec of vibration to reach steady albeit lower level than that obtained during the first part of vibration. The response of the Renshaw cells during the first 100 msec of vibration (phasic response) and that elicited during the last 500 msec of vibration (tonic response) were evaluated for different frequencies of sinusoidal stretch. The mean increase in the firing frequency per imp./sec in the Ia afferents was also calculated using the total one-second period. 3. The response of Renshaw cells to muscle vibration increased with the frequency of vibration and, over the value of 10/sec, appeared to be linearly related to the frequency of the input, at least up to the frequency of 150/sec. Since vibration was of sufficient amplitude to produce driving of all the primary endings of muscle spindles, the responses were expressed as mean increases in the discharge rate of Renshaw cells per average impulse/sec in the Ia afferents. The discharge of the Renshaw cell increased on the average by 2.90 and 1.08 imp./sec per each imp./sec in the Ia afferents during the phasic and the tonic component of the response respectively, while the response calculated during the whole period of vibration corresponded on the average to 1.45 imp./sec per each imp./sec in the Ia afferents. 4. The Renshaw cells tested above responded also with increasing frequencies of discharge to increasing levels of static extension of the GS muscle. In particular the discharge frequency of Renshaw cells was on the average linearly related to muscle extension, at least for values ranging from 0 to 8 mm. The mean increase in discharge rate as a function of the static extension corresponded on the average to 0.89 imp./sec/mm. Since the discharge rate of the primary endings of muscle spindles recorded from the deefferented GS muscle increased by 2.62 imp./sec/mm, it appears that the mean increase in the discharge rate of Renshaw cells as a function of static extension corresponded to 0.34 imp./sec per each imp./sec in the Ia afferents.     

Dispersal of Septoria nodorum Pycnidiospores by Simulated Raindrops in Still Air

Simulated raindrops, diameter c. 3 or 4 mm, fell 13 m down a raintower onto suspensions of Septoria nodorum pycnidiospores, depth 0.5 mm, or infected straw pieces. Splash droplets were collected on pieces of fixed photographic film. It was estimated that one drop generated c. 300 spore carrying splash droplets, containing c. 6000 spores, from a concentrated spore suspension (6.5 × 10⁵ spores/ml) and c. 25 spore-carrying droplets, containing c. 30 spores, from infected straw pieces (11 × 10⁶ spores/g dry wt). When the target was a spore suspension in water without surfactant, most spore-carrying droplets were in the 200—400 μm size category and most spores were carried in droplets with diameter >1000 μm. When surfactant was added to spore suspensions, most spore-carrying droplets were in the 0–200 μm category and most spores were carried in droplets with diameter 200–400 μm and none in droplets >1000 μm. Regression analyses showed a significant (p < 0.001) relationship between square root (number of spores per droplet) and droplet diameter; the slope of the regression line was greatest when surfactant was added to the spore suspensions. The distribution of splash droplets with distance travelled from the target was better fitted by an exponential model than by power law or Gaussian models. The distributions of spore-carrying droplets and spores with distance were fitted better by an exponential model than by a power law model. Thus regressions of log, (number collected) against distance were all significant (p < 0.01); the slopes of the regression lines were steepest when surfactant was added to the spore suspension. At a distance of 10 cm from target spore suspensions most splash droplets and spore-carrying droplets were collected at height 10–20 cm, with none above 40 cm; at a distance of 20 cm there were most at heights 0–10 cm and 40–50 cm.     

Deposition of Erysiphe graminis Conidia on a Barley Crop

Naturally released Erysiphe graminis conidia were trapped (on horizontal slides, on vertical sticky cylinders and in suction traps) in a barley crop infected with powdery mildew and the numbers of single spores and of clumps of different sizes deposited on the traps were counted. The efficiencies of impaction calculated from deposits and wind speed measurements were higher than expected from mean wind speed measurements. The values were consistent with the hypothesis that spores were removedpredominantly in gusts. More than half the conidia were removed in clumps of two or more spores. The measurements suggest that clumps were more effectively deposited than single spores. The measurements demonstrate that spore release mechanisms can influence spore deposition significantly, especially close to the source.