Vegetation type determines spore deposition within a forest–agricultural mosaic landscape

ABSTRACT Predicting fungal community assembly is partly limited by our understanding of the factors driving the composition of deposited spores. We studied the relative contribution of vegetation, geographical distance, seasonality and weather to fungal spore deposition across three vegetation types. Active and passive spore traps were established in agricultural fields, deciduous forests and coniferous forests across a geographic gradient of ∼600 km. Active traps captured the spore community suspended in air, reflecting the potential deposition, whereas passive traps reflected realized deposition. Fungal species were identified by metabarcoding of the ITS2 region. The composition of spore communities captured by passive traps differed more between vegetation types than across regions separated by >100 km, indicating that vegetation type was the strongest driver of composition of deposited spores. By contrast, vegetation contributed less to potential deposition, which followed a seasonal pattern. Within the same site, the spore communities captured by active traps differed from those captured by passive traps. Realized deposition tended to be dominated by spores of species related to vegetation. Temperature was negatively correlated with the fungal species richness of both potential and realized deposition. Our results indicate that vegetation may be able to maintain similar fungal communities across distances, and likely be the driving factor of fungal spore deposition at landscape level.

Laboratory sprayer for testing of microbial biocontrol agents: design and calibration

The lack of commercially available low-cost laboratory spraying equipment for testing microbial control agents can hinder advancement in the field of biocontrol. This study presents an inexpensive, portable sprayer that is calibrated utilizing laboratory consumables. The computer aided design files are made available so that it is freely modifiable and can be used for machine routing or 3D printing. Bioassay data was obtained by spraying Myzus persicae with spores of entomopathogenic fungi. Observed variation in droplet deposition within tested pressure and volume settings, and spore deposition within sprayed concentrations were low. Bioassay results show reproducible mortality for the tested doses.

A Physically Based Theoretical Model of Spore Deposition for Predicting Spread of Plant Diseases

A physically based theory for predicting spore deposition downwind from an area source of inoculum is presented. The modeling framework is based on theories of turbulence dispersion in the atmospheric boundary layer and applies only to spores that escape from plant canopies. A “disease resistance” coefficient is introduced to convert the theoretical spore deposition model into a simple tool for predicting disease spread at the field scale. Results from the model agree well with published measurements of Uromyces phaseoli spore deposition and measurements of wheat leaf rust disease severity. The theoretical model has the advantage over empirical models in that it can be used to assess the influence of source distribution and geometry, spore characteristics, and meteorological conditions on spore deposition and disease spread. The modeling framework is refined to predict the detailed two-dimensional spatial pattern of disease spread from an infection focus. Accounting for the time variations of wind speed and direction in the refined modeling procedure improves predictions, especially near the inoculum source, and enables application of the theoretical modeling framework to field experiment design.

Development of an Aerosol Surface Inoculation Method forBacillusSpores

ABSTRACTA method was developed to depositBacillus subtilisspores via aerosolization onto various surface materials for biological agent decontamination and detection studies. This new method uses an apparatus coupled with a metered dose inhaler to reproducibly deposit spores onto various surfaces. A metered dose inhaler was loaded withBacillus subtilisspores, a surrogate forBacillus anthracis. Five different material surfaces (aluminum, galvanized steel, wood, carpet, and painted wallboard paper) were tested using this spore deposition method. This aerosolization method deposited spores at a concentration of more than 107CFU per coupon (18-mm diameter) with less than a 50% coefficient of variation, showing that the aerosolization method developed in this study can deposit reproducible numbers of spores onto various surface coupons. Scanning electron microscopy was used to probe the spore deposition patterns on test coupons. The deposition patterns observed following aerosol impaction were compared to those of liquid inoculation. A physical difference in the spore deposition patterns was observed to result from the two different methods. The spore deposition method developed in this study will help prepare spore coupons via aerosolization fast and reproducibly for bench top decontamination and detection studies.

Surface Sampling of Spores in Dry-Deposition Aerosols

ABSTRACT The ability to reliably and reproducibly sample surfaces contaminated with a biological agent is a critical step in measuring the extent of contamination and determining if decontamination steps have been successful. The recovery operations following the 2001 attacks with Bacillus anthracis spores were complicated by the fact that no standard sample collection format or decontamination procedures were established. Recovery efficiencies traditionally have been calculated based upon biological agents which were applied to test surfaces in a liquid format and then allowed to dry prior to sampling tests, which may not be best suited for a real-world event with aerosolized biological agents. In order to ascertain if differences existed between air-dried liquid deposition and biological spores which were allowed to settle on a surface in a dried format, a study was undertaken to determine if differences existed in surface sampling recovery efficiencies for four representative surfaces. Studies were then undertaken to compare sampling efficiencies between liquid spore deposition and aerosolized spores which were allowed to gradually settle under gravity on four different test coupon types. Tests with both types of deposition compared efficiencies of four unique swabbing materials applied to four surfaces with various surface properties. Our studies demonstrate that recovery of liquid-deposited spores differs significantly from recovery of dry aerosol-deposited spores in most instances. Whether the recovery of liquid-deposited spores is overexaggerated or underrepresented with respect to that of aerosol-deposited spores depends upon the surface material being tested.

Variation in Rates of Spore Deposition of Fusarium circinatum, the Causal Agent of Pine Pitch Canker, Over a 12-Month-Period at Two Locations in Northern California

Patterns of spore deposition by Fusarium circinatum, the causal agent of pine pitch canker (PPC) of Monterey pine (Pinus radiata) and other conifers, were studied between May 2003 and April 2004 at two sites in Northern California using a novel spore trapping method combined with a real-time polymerase chain reaction (PCR) approach. At each study site, two plots were sampled by placing spore traps at 100 m intervals along transects 600 m in length. The air was sampled continuously by exchanging the spore traps every 2 weeks. The spore deposition rate (DR), ranged from 0 to 1.3 × 105 spores m–2. Spores were detected throughout the year, with higher trapping frequencies (TF) during the rainy season (November to April), than during the dry season (May to October). The detection of spores on traps at distances larger than 200 m from any Monterey pine, suggests at least midrange aerial dispersal. Finally, different inoculum loads were associated with trees displaying different levels of disease symptoms, suggesting infectiousness of the pathogen varies as the disease progresses. This study represents one of the first documenting continuous inoculum pressure values over an entire year for a forest pathogen, and provides important epidemiological information that will be invaluable in the development of disease progression models.