Dispersal of Rhynchosporium secalis conidia from infected barley leaves or straw by simulated rain

Simulated rain (mean drop diameter c. 1 or 3 mm) was allowed to fall for 10 – 15 min on to barley leaves or straw infected by Rhynchosporium secalis (leaf blotch). The leaves were supported on a mesh through which run-off water drained and the straw was supported on a rigid surface on which run-off water collected. The numbers of R. secalis conidia and spore-carrying splash droplets collected by horizontal samplers (microscope slides and pieces of photographic film) decreased rapidly with increasing distance from and increasing height above the sources, with half-distances of 2 – 10 cm. Less than 10% of the spores or droplets reached heights of more than 30 cm. Incident drops 3 mm in diameter produced more spore-carrying droplets and dispersed more conidia than did 1 mm drops. The size category of splash droplets with the greatest proportion of the spore-carrying droplets dispersed by 3 mm drops was 200 – 400 μm, whether the source was infected barley leaves or barley straw. For leaves or straw the greatest proportions of spores were carried in droplets > 1000 μm in diameter. The mean diameter of spore-carrying droplets (478 μm) dispersed from free-draining leaves was less than that of droplets from straw plus run-off water (563 μm). However, the leaf source had more spores cm^-2 and the mean number of spores per droplet was greater (113 as opposed to 6·8) than for the straw source.     

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.     

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.     

Patterns of unobstructed splash dispersal

Unobstructed splash dispersal patterns were measured in the absence of rain over mown grass using a fluorescent tracer, and a colorimetric method was used indoors in still air. When drops fell into a thin horizontal water film 0–1 mm deep, the volume of the incident drops dissipated as splash droplets was similar to the volume splashed from the film, irrespective of the distance of fall of the drops. Drop size, angle of inclination and distance of fall had significant effects on the volume of drops splashed from an inclined surface. The effects of rigidity, inclination and nature of surface were found to be significant when drops impacted onto surfaces with or without a wax covering and either rigidly or loosely supported. When splash- and dry-air-dispersed Lycopodium spores were simultaneously released, many more splashed spores were caught close to the source, but the dispersal gradient of splashed spores was steeper than that of dry-air-dispersed spores. Splash-dispersed spores were caught on slides, cylinders and rotorods but trap efficiency could not be evaluated.     

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.     

A photographic technique for investigating the splashing of water drops on leaves

The formation of splash from water drops falling onto bean leaves was investigated by multiple exposure flash photography. A relatively simple technique was developed which recorded up to four images at 2 ms intervals, of a splash on one photograph. The images were recorded on colour reversal film, and the flash sequence was colour coded to aid the interpretion of the superimposed images. Analysis of the photographs allowed measurement of the initial speed and angle of ejection of different sized splash droplets. Droplets were ejected at speeds of up to about 10 m s^-1although most speeds were less than about 3 m s^-1, and small droplets tended to travel faster than large ones. Droplets were ejected at all angles but the majority were ejected almost horizontally; there was little correlation between droplet size and angle of ejection. Deposition patterns estimated from calculated droplet trajectories compared well with measured ones.     

Myxosoma microspora n. sp. (Myxosporidia: Protozoa) parasitic in the gills of Mugil cephalus L.

A new histozoic species of myxosporidian, Myxosoma microspora n.sp., infecting the gill filaments of Mugil cephalus is described. Cysts measuring 0.5–1.0 mm in diameter were found attached to the gill filaments. Spherical or slightly oval, spores 4.8–5.2 μm in diameter, were present and possessed a thin outer mucous envelope which appeared as small conical protuberances at the ends of the equatorial axis. Polar capsules were pyriform in shape, equal in size and measured 1.6–2.0 × 1.0–1.2 μm; the polar filaments were 22–28 μm in length. There was a bean-shaped sporoplasm measuring 3.5 × 1.5 μm. No iodine vacuole was detected when the parasite was stained with Lugol's iodine.     

Splash dispersal of fungus spores and fungicides in the laboratory and greenhouse

A laboratory technique is described for the production of drops of simulated rain in which fungal spores were suspended. When such drops containing conidia of Botrytis fabae impacted on a target leaf the secondary droplets produced infections on receptor broad bean leaves. The capacity of fungicides applied to the target leaf to redistribute in secondary splash droplets was examined in terms of the infectivity of the spores in the droplets. The extent to which a copper fungicide reduced infection on the receptor leaves was related to the level and tenacity of the fungicide deposit on the target leaf. The effect of wetting agents on the redistribution of this fungicide could probably be explained by their influence on the tenacity of the initial deposit. In general the capacity of different fungicides to inhibit infection by the secondary droplets was related to the inherent toxicity of the fungicides to B. fabae. Implications of the dispersal of spores and fungicides by rain splash are briefly considered with reference to field conditions.     

Splash dispersal of Coniothyrium minitans in the glasshouse

Coniothyrium minitans, a mycoparasite with biocontrol activity against Sclerotinia sclerotiorum, was found to disperse during glasshouse trials where overhead irrigation was used. Consequently, the role of water splash in dispersal of C. minitans was investigated using soil-incorporated inoculum and a range of irrigation regimes found to occur in the glasshouse. The resulting inoculum deposition over horizontal distances up to 2 m was measured. Using drops < 6 mm diameter at 680 mm h^-1, C. minitans was splash-dispersed at least 2.0 m, whereas with drops > 6 mm diameter at 30 mm h^-1 it was dispersed to only 1.75 m. Irrigation with droplets < 1mm diameter at 49 mm h^-1 failed to disperse inoculum beyond 0.5 m. The dispersal gradient produced by drops < 6 mm diameter at 680 mm h^-1 was best described mathematically by the power function, whereas irrigation with drops > 6 mm diameter at 30 mm h^- resulted in a gradient described well by power or exponential functions. The latter regime produced a significantly steeper gradient than irrigation with drops < 6 mm diameter at 680 mm h^-1. C. minitans was isolated using an Andersen air sampler at concentrations of 2839 cfu m^-3 or 22 cfu m^-3 during irrigation with drops < 6 mm diameter at 680 mm h^-1 or > 6 mm diameter at 30 mm h^-1, respectively. After irrigation, deposition of C. minitans-canying aerosol particles declined exponentially and distance from source had no effect on the amount of inoculum isolated. Conidia of C. minitans, splash-dispersed by irrigation with drops < 6 mm diameter at 680 mm h^-1 were able to infect sclerotia of S. sclerotiorum such that almost all sclerotia at 0.5 m from the inoculum source, and c. 50% of those at 2.0 m, became infected with the mycoparasite.     

Mushrooms as Rainmakers: How Spores Act as Nuclei for Raindrops

Millions of tons of fungal spores are dispersed in the atmosphere every year. These living cells, along with plant spores and pollen grains, may act as nuclei for condensation of water in clouds. Basidiospores released by mushrooms form a significant proportion of these aerosols, particularly above tropical forests. Mushroom spores are discharged from gills by the rapid displacement of a droplet of fluid on the cell surface. This droplet is formed by the condensation of water on the spore surface stimulated by the secretion of mannitol and other hygroscopic sugars. This fluid is carried with the spore during discharge, but evaporates once the spore is airborne. Using environmental electron microscopy, we have demonstrated that droplets reform on spores in humid air. The kinetics of this process suggest that basidiospores are especially effective as nuclei for the formation of large water drops in clouds. Through this mechanism, mushroom spores may promote rainfall in ecosystems that support large populations of ectomycorrhizal and saprotrophic basidiomycetes. Our research heightens interest in the global significance of the fungi and raises additional concerns about the sustainability of forests that depend on heavy precipitation.     

Sphaerospora epinepheli N. Sp. (Myxosporea: Sphaerosporidae) Observed in Grouper (Epinephelus malabaricus)

Sphaerospora epinepheli n. sp. is described from grouper, Epinephelus malabaricus , in cage-cultured and wild fish collected from both coastal lines of southern Thailand. Subspherical to spherical spores and mono- or disporous pseudoplasmodia were observed in the lumen of kidney tubules. Pseudoplasmodia were round to elongate, size range 15.6–22.9 μm (length) × 8.4–21.6 μm (width). Spores were 7.8–10.0 μm (length) × 12.3–14.5 μm (thickness), and 7.0–9.5 μm (width) with two spherical polar capsules of equal size measuring 2.9–4.4 μm in diameter and containing polar filaments with six or seven windings. Two uninucleate sporoplasms showed iodine vacuoles. Blood stages, similar to C-blood protozoans observed from freshwater fish in Europe, were found from peripheral blood smears of grouper. Ultrastructural studies of blood stages showed a similar structure to unidentified mobile protozoans from the blood of carp. Electron dense bodies were observed in the cytoplasm of the primary cell blood stages. Infected proximal-tubular epithelial cells showed highly vacuolated cytoplasm and pycnotic nuclei.     

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.     

Measurement of light gradients and spectral regime in plant tissue with a fiber optic probe

A method is described in which light gradients and spectral regime can be measured within plant tissue using fiber optics. A fiber optic probe was made by modifying a single optical fiber (200 μm diameter) so that it had a light harvesting end that was a truncated tip 20–70 μm in diameter. The probe was a directional sensor with a half-band acceptance angle of 17–20°. Light measurements were made as the fiber optic probe was driven through plant tissue by a motorized micromanipulator, and the light that entered the fiber tip was piped to a spectroradiometer. By irradiating green leaf tissue of the succulent Crassula falcata L. with collimated light and inserting the probe from different directions, it was possible to measure light quality and quantity at different depths. Collimated light was scattered completely by the initial 1.0 mm of leaf tissue, which also greatly attenuated all light except the green and far-red. Light scatter contributed significantly to light quantity and had a pronounced spectral structure. Immediately beneath the irradiated surface the amount of light at 550 nm was 1.2 times that of the incident light. The light gradient declined rapidly to 0.5 times incident light at 1.4 mm depth. In contrast, the amount of light at 750 nm increased during the initial 0.5 mm to 2.9 times incident light and then declined linearly to 0.5 times incident light at the dark side of the leaf (4.5 mm). The implications of the magnitude of the contribution of light scatter to the light gradient is also discussed.     

Estimation of the spectrum of the aerial dispersion of Bacillus thuringiensis

The present paper estimates the number of viable spores of Bacillus thuringiensis per droplet and reveals the importance of this data to improve B. thuringiensis treatments. For a given diameter, droplets from Futura formula contained two times more spores than droplets from the formulas used to date. It is preferable to have B. thuringiensis droplets larger than those of chemical insecticides. A Grumman AgCat aircraft calibrated to give the desired larger droplets resulted in successful B. thuringiensis treatments.     

Collection characteristics of a batch-type wetted wall bioaerosol sampling cyclone

The collection efficiency and sample retention of a batch-type wetted wall bioaerosol sampling cyclone (BWWC) were experimentally characterized. The BWWC is designed to sample air at 400 l/min and concentrate the particles into 12 ml of water. Aerosol is transported into a cylindrically-shaped axial flow cyclone through a tangential slot and the particles are impacted on the inner wall, which is wetted by air shear acting on a liquid pool at the base of the cyclone. The retention of collected particles and the aerosol collection efficiency of the BWWC were evaluated with polystyrene latex beads (PSL), sodium fluorescein/oleic acid droplets, and Bacillus atrophaeus (aka BG) spores. The retention of particles was determined by adding hydrosol directly into the device, running the BWWC for a pre-set period of time, and then determining the amount of particulate matter recovered relative to the initial amount. For 1-μm diameter PSL, 90% of the particles were recoverable from the cyclone body immediately after their introduction; however, only 10% were retained in the collection liquid after 8 h of operation. The aerosol sampling efficiency was determined by comparing the amount of particulate matter collected in the liquid with that collected by a reference filter. The collection efficiency was 50–60% for 1- and 3-μm polystyrene (PSL) particles, and 1.5% for 10-μm oleic acid particles. The efficiency for 3-μm oleic acid droplets was 35%. Explanations are provided for the difference between liquid and solid particle behavior, and for the low efficiency for the large liquid particles. The collection efficiency for single spore BG was slightly lower than that for 1-μm PSL.     

Spawning of Basilichthys microlepidotus (Jenyns)

Characteristics of the reproductive cycle of Basilichthys microlepidotus (Jenyns), an atherinid fish living in a South American Mediterranean arid zone, are analysed. Specimens 9-170 mm in total length were captured from August 1982 to April 1984 in the Rio Choapa, in northern Chile (31%%45' S, 71°15' W). Analysis of gonadosomatic index fluctuations, and determination of the proportion of mature males and females captured during the study period, allowed the estimation of the duration of the reproductive cycle.
Females 70–170 mm in total length contained 2000–9000 intraovarian oocytes, of which approximately 80% were vitellogenic (100–1300 μm in diameter) and exhibited a polymodal distribution. A further 4 to 15% of the total oocytes, measured 1700–2000 μm in diameter and appeared ready to be spawned. In 30% of the examined ovaries some of the largest oocytes exhibited signs of degeneration, or atresia. Juveniles approximately 30 mm in total length were captured over most of the study period. Results strongly suggest that B. microlepidotus has multiple spawnings.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

Light and Electron Microscopy of the Spore of Myxobolus heckelii n. sp. (Myxozoa), Parasite from the Brazilian Fish Centromochlus heckelii (Teleostei, Auchenipteridae)

ABSTRACT. A myxosporean parasitizing the gill filaments of the freshwater teleost fish Centromochlus heckelii collected in the Tocantins River (Lower Amazonian Region, Brazil) is described using light and electron microscopy. This parasite produces spherical to ellipsoidal cyst-like plasmodia up to 250 μm in diameter, with a thick wall strengthened by several stratified juxtaposed crossed collagen layers, whose thickness varies according to the number of the layers. Several compressed fibroblasts are observed among the collagen fibrils. Deposits of spherical dense material are scattered at the internal periphery of the cysts. Plasmodia and different developmental stages, including immature and mature spores, filled the central region of the cysts. The spore body is ellipsoidal in valvar view and biconvex in sutural view. It is formed by two equal-sized and symmetric valves measuring 12.7 μm long (12.2–13.1) ( n =50), 6.6 μm wide (6.3–6.9) ( n =25), and 4.0 μm (3.7–4.4) ( n =20) thick. A thin layer formed by fine and anastomosed microfibrils is observed at the spore surface. Two equal, elongated pyriform polar capsules measure 2.9 μm (2.7–3.3) × 1.7 μm (1.4–2.0) ( n =25), each containing four or five oblique polar filament coils. The binucleated sporoplasm contains numerous spherical sporoplasmosomes, glycogen particles, and a large vacuole with fine granular matrix. Based on the morphological and ultrastructural differences and specificity of the host, we describe this isolate as a new myxosporidian, Myxobolus heckelii n. sp. (Myxozoa, Myxosporea).     

Resource utilization by the freshwater deposit feeder Ptychoptera townesi (Diptera: Ptychopteridae)

SUMMARY 1. False crane fly larvae, Ptychoptera townesi (Diptera), occurred in high densities in a flow-controlled section of stream where fine particulate organic matter (FPOM; 0.45 μm to 1 mm in diameter) had accumulated, but were quite rare both upstream and downstream from the section.
2. In laboratory studies, P. townesi grew only on FPOM less than 250 μm. Larvae consistently grew fastest when fed small particles (0.45–53 μm in diameter).
3. Ptychoptera townesi consumed relatively small amounts (0.002 mg per mg animal dry mass day⁻¹) of FPOM (0.45–53 μm). They had long gut content passage times (greater than 19 h) and relatively high efficiencies of conversion of ingested food to body substance (20.7%). Gut content passage times were variable, and depended partially on the nature of the substrate.
4. False crane fly larvae compacted FPOM into faecal pellets considerably larger in size than particles ingested. They lost mass when allowed to feed on their own faecal material, as well as on faeces greater than 250 μm in diameter produced by shredders. However, they survived and grew on shredder faeces (53–500 μm in diameter) that contained a mixture of smaller particles and particles too large for ingestion.
5. The overall pattern of resource utilization by P. townes involved slow handling of relatively small volumes of food, which probably passed once only through a complex alimentary tract.     

Ultrastructural aspects of the cytoplasmic origin and accumulation of oil in olive fruit (Olea europaea)

The development of oil bodies and oil droplets in fruits of olive was examined at the ultrastructural level. Both oil bodies that form in young fruits and oil droplets that develop with fruit maturation are cytoplasmic bodies. The formation of the small oil bodies occurs in localized regions of the cytoplasm. These bodies are closely associated and fuse together, forming a small oil droplet that protruded against and indented the vacuolar membrane. As the fruit matures, new oil bodies appear to form and fuse with the oil droplet, resulting in the formation of a single large oil droplet of about 30 μm in diameter in most mature mesocarp cells. The cytoplasmic region where the oil bodies formed had a granulate, ultrastructural appearance, and cytoplasmic components such as membranes and ribosomes were noticeably absent in these regions. The granulate material coated the oil bodies and oil droplets, and appeared as a thin, compressed band between the round inner surface of the droplets and the indented tonoplast. We suggest that this granulate material is involved in the synthesis of the oil and, with enlargement of the oil bodies, this coat becomes thinner in regions where they are closely associated, resulting in zones where confluence of the oil occurs.