Microbial vectoring capacity by internal- and external-infesting stored product insects after varying dispersal periods between novel food patches: An underestimated risk.

Understanding the ability of internal- and external-infesting stored product insects to vector microbes is important for estimating the relative risk that insects pose to postharvest commodities as they move between habitat patches and in the landscape. Thus, the aim of the current study was to evaluate and compare the microbial growth in novel food patches at different dispersal periods by different populations of Sitophilus oryzae (e.g., internal-infesting) and Lasioderma serricorne (e.g., external-infesting). Adults of both species collected from laboratory colonies or field-captured populations were either placed immediately in a novel food patch, or given a dispersal period of 24 or 72 h in a sterilized environment before entering a surrogate food patch. Vectored microbes in new food patches were imaged after 3 or 5 days of foraging, and microbial growth was processed using ImageJ while fungal species were identified through sequencing the ITS4/5 ribosomal subunit. We found that increasing dispersal time resulted in multiple-fold reductions in microbial growth surrogate food patches by L. serricorne but not S. oryzae. This was likely attributable to higher mobility by S. oryzae than L. serricorne. A total of 20 morphospecies were identified from 13 genera among the 59 sequences, with a total of 23% and 16% classified as Aspergillus and Penicillium spp. Our data suggest that there is a persistent risk of microbial contamination by both species, which has important food safety implications at food facilities.

Density-mediated foraging behavioral responses of Rhyzopertha dominica (Coleoptera: Bostrichidae) and Tribolium castaneum (Coleoptera: Tenebrionidae).

Tribolium castaneum and Rhyzopertha dominica are cosmopolitan, destructive postharvest pests. Although research has investigated how high densities of T. castaneum affect attraction to the aggregation pheromone by conspecifics, research into the behavioral response of both species to food cues after high density exposure has been lacking despite its importance to foraging ecology. Our goal was to manipulate and observe the effects of crowding on the behavioral response of both species to common food and pheromonal stimuli and to determine how the headspace emission patterns from grain differed under increasing densities. Densities of colonies for both species was altered (10-500 adults) on a fixed quantity of food (10 g of flour or whole wheat), then the behavioral response to common food and pheromonal cues was evaluated in a wind tunnel and release-recapture experiment, while volatiles were examined through gas chromatography coupled with mass spectrometry. Importantly, at least for T. castaneum, crowded conditions attenuate attraction to food-based stimuli, but not pheromonal stimuli. Crowding seemed to have no effect on R. dominica attraction to food and pheromonal stimuli in the wind tunnel, but exposure to high density cues did elicit 2.1-3.8-fold higher captures in traps. The relative composition and abundance of headspace volatiles emitted varied significantly with different densities of beetles and was also species-specific. Overall, our results have implications for expanding our understanding of the foraging ecology of two economically important pests.

Attraction, mobility, and preference by Lasioderma serricorne (Coleoptera: Ptinidae) to microbially-mediated volatile emissions by two species of fungi in stored grain.

Insects and microbes are known to interact in a variety of ways at food facilities, compounding damage. However, little research has explicated how specific common fungal species affect the behavior of the cosmopolitan secondary stored product pest, Lasioderma serricorne. Enhanced knowledge about attraction to microbially-produced volatile organic compounds (MVOCs) may be used to manipulate insect behavior. Aspergillus flavus and Fusarium verticillioides are two common, widespread pre- and postharvest fungi on small cereals that produce aflatoxins and fumonisins, respectively, while directly competing with each other for nutrients. Our goals were to (1) characterize the volatile emissions from grain inoculated by A. flavus or F. verticillioides derived from the cuticle of L. serricorne compared to uninoculated and sanitized grain, and (2) understand how MVOCs from each fungal species affects mobility, attraction, and preference by L. serricorne. Headspace collection revealed that the F. verticillioides- and A. flavus-inoculated grain produced significantly different volatiles compared to sanitized grain or the positive control. Changes in MVOC emissions affected close-range foraging during an Ethovision movement assay, with a greater frequency of entering and spending time in a small zone with kernels inoculated with A. flavus compared to other treatments. In the release-recapture assay, MVOCs were found to be attractive to L. serricorne at longer distances in commercial pitfall traps. There was no preference shown among semiochemical stimuli in a still-air, four-way olfactometer. Overall, our study suggests that MVOCs are important for close- and long-range orientation of L. serricorne during foraging, and that MVOCs may have the potential for inclusion in behaviorally-based tactics for this species.

Grain Inoculated with Different Growth Stages of the Fungus, Aspergillus flavus, Affect the Close-Range Foraging Behavior by a Primary Stored Product Pest, Sitophilus oryzae (Coleoptera: Curculionidae).

Although some research has investigated the interactions among stored product insects and microbes, little research has examined how specific fungal life stages affect volatile emissions in grain and linked it to the behavior of Sitophilus oryzae, the cosmopolitan rice weevil. Thus, our goals were to 1) isolate, culture, and identify two fungal life stages of Aspergillus flavus, 2) characterize the volatile emissions from grain inoculated by each fungal morphotype, and 3) understand how microbially-produced volatile organic compounds (MVOCs) from each fungal morphotype affect foraging, attraction, and preference by S. oryzae. We hypothesized that the headspace blends would be unique among our treatments and that this will lead to preferential mobility by S. oryzae among treatments. Using headspace collection coupled with GC-MS, we found the sexual life stage of A. flavus had the most unique emissions of MVOCs compared to the other semiochemical treatments. This translated to a higher interaction with kernels containing grain with the A. flavus sexual life stage, as well as a higher cumulative time spent in those zones by S. oryzae in a video-tracking assay in comparison to the asexual life stage. While fungal cues were important for foraging at close-range, the release-recapture assay indicated that grain volatiles were more important for attraction at longer distances. There was no significant preference between grain and MVOCs in a four-way olfactometer. Overall, this study enhances our understanding of how fungal cues affect the close and longer range foraging ecology of a primarily stored product insect.

A Systematic Review of the Behavioral Responses by Stored-Product Arthropods to Individual or Blends of Microbially Produced Volatile Cues.

Microbes are ubiquitous and play important ecological roles in a variety of habitats. While research has been largely focused on arthropods and microbes separately in the post-harvest supply chain, less attention has been paid to their interactions with each other. Up to this point, there has been no attempt to systematically describe the patterns of behavioral responses by stored-product insects to microbially produced volatile organic compounds (MVOCs). Thus, our aims were to evaluate whether stored-product arthropods were primarily and significantly attracted, repelled, or had a net neutral effect (e.g., unaffected or mixed) by MVOCs presented as (1) complex headspace blends or (2) single constituents and known mixtures. In total, we found 43 articles that contained 384 sets of tests with different combinations of methodology and/or qualitative findings, describing the behavioral responses of 24 stored-product arthropod species from two classes, four orders, and 14 families to 58 individual microbial compounds and the complex headspace blends from at least 78 microbial taxa. A total of five and four stored-product arthropod species were significantly attracted and repelled by MVOCs across odor sources, respectively, while 13 were unaffected or exhibited mixed effects. We summarize the biases in the literature, including that the majority of tests have occurred in the laboratory with a limited subset of methodology and has largely only assessed the preference of adult arthropods. Finally, we identify foundational hypotheses for the roles that MVOCs play for stored-product arthropods as well as gaps in research and future directions, while highlighting that the behavioral responses to MVOCs are complex, context-, and taxon-dependent, which warrants further investigation.