Exposure of Larvae of the Solitary Bee Osmia bicornis to the Honey Bee Pathogen Nosema ceranae Affects Life History

Wild bees are important pollinators of wild plants and agricultural crops and they are threatened by several environmental stressors including emerging pathogens. Honey bees have been suggested as a potential source of pathogen spillover. One prevalent pathogen that has recently emerged as a honey bee disease is the microsporidian Nosema ceranae. While the impacts of N. ceranae in honey bees are well documented, virtually nothing is known about its effects in solitary wild bees. The solitary mason bee Osmia bicornis is a common pollinator in orchards and amenable to commercial management. Here, we experimentally exposed larvae of O. bicornis to food contaminated with N. ceranae and document spore presence during larval development. We measured mortality, growth parameters, and timing of pupation in a semi-field experiment. Hatched individuals were assessed for physiological state including fat body mass, wing muscle mass, and body size. We recorded higher mortality in the viable-spore-exposed group but could only detect a low number of spores among the individuals of this treatment. Viable-spore-treated individuals with higher head capsule width had a delayed pupation start. No impact on the physiological status could be detected in hatched imagines. Although we did not find overt evidence of O. bicornis infection, our findings indicate that exposure of larvae to viable N. ceranae spores could affect bee development.

Combined Effects of Carbonation with Heating and Fatty Acid Esters on Inactivation and Growth Inhibition of Various Bacillus Spores

The effects of carbonation treatment (1 to 5 MPa, 30 min) plus heat treatment (30 to 80°C, 30 min) in the presence of various fatty acid esters (FAEs; 0.05 and 0.1%, wt/vol) on counts of viable Bacillus subtilis spores were investigated. FAEs or carbonation alone had no inactivation or growth inhibition effects on B. subtilis spores. However, carbonation plus heat (CH; 80°C, 5 MPa, 30 min) in the presence of mono- and diglycerol fatty acid esters markedly decreased counts of viable spores, and the spore counts did not change during storage for 30 days. The greatest decrease in viable spore counts occurred in the presence of monoglycerol fatty acid esters. Under CH conditions, inactivation and/or growth inhibition occurred at only 80°C and increased with increasing pressure. The greatest decrease in spore counts (more than 4 log units) occurred with CH (80°C, 5 MPa, 30 min) in the presence of monoglycerol fatty acid esters. However, this treatment was less effective against Bacillus coagulans and Geobacillus stearothermophilus spores.

Evaluation of methods for long term storage of the boysenberry downy mildew pathogen Peronospora sparsa

Downy mildew of boysenberry is caused by the biotrophic pathogen Peronospora sparsa To ensure supplies of viable spore inoculum for infection experiments six storage methods were assessed the leaf discs cut from sporulating areas of the leaf stored dry or in 20 glycerol and spores were suspended in 20 glycerol all three of which were stored at either 20C or 80C After 1 2 4 or 6 months storage spore viability and the capability to infect leaf discs were evaluated Storage methods had no significant effects on spore germination or infection Storage time and temperature significantly (P

Identification by Quantitative Carrier Test of Surrogate Spore-Forming Bacteria To Assess Sporicidal Chemicals for Use against Bacillus anthracis

ABSTRACT The spores of six strains of Bacillus anthracis (four virulent and two avirulent) were compared with those of four other types of spore-forming bacteria for their resistance to four liquid chemical sporicides (sodium hypochlorite at 5,000 ppm available chlorine, 70,000 ppm accelerated H2O2, 1,000 ppm chlorine dioxide, and 3,000 ppm peracetic acid). All test bacteria were grown in a 1:10 dilution of Columbia broth (with manganese) incubated at 37°C for 72 h. The spore suspensions, heat treated at 80°C for 10 min to rid them of any viable vegetative cells, contained 1 × 108 to 3 × 108 CFU/ml. The second tier of the quantitative carrier test (QCT-2), a standard of ASTM International, was used to assess for sporicidal activity, with disks (1 cm in diameter) of brushed and magnetized stainless steel as spore carriers. Each carrier, with 10 μl (≥106 CFU) of the test spore suspension in a soil load, was dried and then overlaid with 50 μl of the sporicide being evaluated. The contact time at room temperature ranged from 5 to 20 min, and the arbitrarily set criterion for acceptable sporicidal activity was a reduction of ≥106 in viable spore count. Each test was repeated at least three times. In the final analysis, the spores of Bacillus licheniformis (ATCC 14580T) and Bacillus subtilis (ATCC 6051T) proved to be generally more resistant than the spores of the strains of B. anthracis tested. The use of one or both of the safe and easy-to-handle surrogates identified here should help in developing safer and more-effective sporicides and also in evaluating the field effectiveness of existing and newer formulations in the decontamination of objects and surfaces suspected of B. anthracis contamination.

Spatial Patterns of Viable Spore Deposition of Gibberella zeae in Wheat Fields

An increased understanding of the epidemiology of Gibberella zeae will contribute to a rational and informed approach to the management of Fusarium head blight (FHB). An integral phase of the FHB cycle is the deposition of airborne spores, yet there is no information available on the spatial pattern of spore deposition of G. zeae above wheat canopies. We examined spatial patterns of viable spore deposition of G. zeae over rotational (lacking cereal debris) wheat fields in New York in 2002 and 2004. Viable, airborne spores (ascospores and macroconidia) of G. zeae were collected above wheat spikes on petri plates containing a selective medium and the resulting colonies were counted. Spores of G. zeae were collected over a total of 68 field environments (three wheat fields during 54 day and night sample periods over 2 years) from spike emergence to kernel milk stages of local wheat. Spatial patterns of spore deposition were visualized by contour plots of spore counts over entire fields. The spatial pattern of spore deposition was unique for each field environment during each day and night sample period. Spore deposition patterns during individual sample periods were classified by spatial analysis by distance indices (SADIE) statistics and Mantel tests. Both analyses indicated that the majority (93%) of the spore deposition events were random, with the remainder being aggregated. All of the aggregated patterns were observed during the night. Observed patterns of spore deposition were independent of the mean number of viable spores deposited during individual sample periods. The spatial pattern for cumulative spore deposition during anthesis in both years became aggregated over time. Contour maps of daily and cumulative spore deposition could be compared with contour maps of FHB incidence to gain insights into inoculum thresholds and the timing of effective inoculum for infection.