Examining historical rates of leafcutting bee cell pathogens to establish baseline infectivity rates for alfalfa seed growers

The alfalfa leafcutting bee (Megachile rotundata) is one of the primary pollinators for the alfalfa seed industry. The alfalfa leafcutting bee is a solitary cavity nesting bee. Female Megachile rotundata bees will construct and provision individual brood cells lined with cut leaves (cocoon) and will gather nectar and pollen to place within the constructed cocoon. The female bee will lay a single egg within the constructed cocoon and leave the egg to undergo larval stage development and pupation into the adult stage. During this time multiple pathogens and parasitoids can prey on the developing larvae, resulting in the loss of the future adult bee. A major concern for commercial alfalfa seed growers is the presence of invertebrate pests and fugal pathogens. In the present study, we used historical data from the Parma Cocoon Diagnostic Laboratory to determine baseline rates of pathogen and parasite infection of Megachile rotundata cells and used this analysis to determine historical infection rates and cutoffs for management practices. Additionally, using a Faxitron (X-ray) analysis for Megachile rotundata cell obtained in 2020, we compared the presence of chalkbrood, pathogens, and parasitoids in samples collected from both growers stocks and newly purchased Canada bees. The results of the investigation demonstrate historical averages of the presence of chalkbrood, pathogens, and parasitoids. We also show a significant increase in chalkbrood and predators in 2007-2011 and a significant difference in chalkbrood and predators between bee samples obtained from Canada and grower stocks.

Multiplex PCR reveals unique trends in pathogen and parasitoid infestations of alfalfa leafcutting brood cells.

The alfalfa leafcutting bee Megachile rotundata Fabricius (HYMENOPTERA: Megachilidae) is an important pollinator for multiple agricultural seed commodities in the United States. Megachile rotundata is a solitary bee that forms brood cocoons where its larvae can develop. During the developmental stages of growth, broods can be preyed upon by multiple different fungal and bacterial pathogens and insect predators and parasitoids, resulting in the loss of the developing larvae. Larval loss is a major concern for alfalfa (Medicago sativa L.) seed producers because they rely on pollinator services provided by Megachile rotundata and reduced pollination rates result in lower yields and increased production costs. In the present study, we examined the taxonomic composition of organisms found within M. rotundata brood cells using a multiplex PCR assay which was developed for the detection of the most common bacterial, fungal, and invertebrate pests and pathogens of M. rotundata larvae. Known pests of M. rotundata were detected, including members of the fungal genus Ascosphaera, the causative agent of chalkbrood. Co-infection of single brood cells by multiple Ascosphaera species was confirmed, with potential implications for chalkbrood disease management. The multiplex assay also identified DNA from more than 2,400 total species including multiple new predators and pathogenetic species not previously documented in associated with M. rotundata brood cells.

Effect of Insecticides on Natural-Enemies

Pesticides management options for control of invertebrate pests in many parts of the world. Despite an increase in the use of pesticides, crop losses due to pests have remained largely unchanged for 30–40 years. Beyond the target pests, broad-spectrum pesticides may affect non-target invertebrate species, including causing reductions in natural enemy population abundance and activity, and competition between pest species. Assays of invertebrates against weathered residues have shown the persistence of pesticides might play an important part in their negative impacts on natural enemies in the field. A potential outcome of frequent broad-spectrum pesticide use is the emergence of pests not controlled by the pesticides but benefiting from reduced mortality from natural enemies and competitive release, commonly known as secondary pests.

Organic Control Strategies for Use in IPM of Invertebrate Pests in Apple and Pear Orchards

Growers of organic tree fruit face challenges in controlling some pests more easily suppressed by broad-spectrum insecticides in conventionally managed orchards. In recent decades, there has been a move towards organically growing varieties normally reliant on synthetic chemical pesticides (e.g., Gala), often to meet retailer/consumer demands. This inevitably makes crop protection in organic orchards more challenging, as modern varieties can be less tolerant to pests. In addition, there have been substantial reductions in plant protection product (PPP) approvals, resulting in fewer chemical options available for integrated pest management (IPM)-maintained orchards. Conversely, the organic management of fruit tree pests involves many practices that could be successfully implemented in conventionally grown crops, but which are currently not. These practices could also be more widely used in IPM-maintained orchards, alleviating the reliance on broad-spectrum PPP. In this review, we evaluate organic practices, with a focus on those that could be incorporated into conventional apple and pear production. The topics cover cultural control, biological control, physical and pest modifications. While the pests discussed mainly affect European species, many of the methods could be used to target other global pests for more environmentally sustainable practices.

Asexual Epichloë Fungi—Obligate Mutualists

Asexual Epichloë are obligate fungal mutualists that form symbiosis with many temperate grass species, providing several advantages to the host. These advantages include protection against vertebrate and invertebrate herbivores (i.e., grazing livestock and invertebrate pests, respectively), improved resistance to phytopathogens, increased adaptation to drought stress, nutrient deficiency, and heavy metal-containing soils. Selected Epichloë strains are utilised in agriculture mainly for their pest resistance traits, which are moderated via the production of Epichloë-derived secondary metabolites. For pastoral agriculture, the use of these endophyte infected grasses requires the balancing of protection against insect pests with reduced impacts on animal health and welfare.

A Rapid and Cost-Effective Identification of Invertebrate Pests at the Borders Using MinION Sequencing of DNA Barcodes

The rapid and accurate identification of invertebrate pests detected at the border is a challenging task. Current diagnostic methods used at the borders are mainly based on time consuming visual and microscopic examinations. Here, we demonstrate a rapid in-house workflow for DNA extraction, PCR amplification of the barcode region of the mitochondrial cytochrome oxidase subunit I (COI) gene and Oxford Nanopore Technologies (ONT) MinION sequencing of amplified products multiplexed after barcoding on ONT Flongle flow cells. A side-by-side comparison was conducted of DNA barcode sequencing-based identification and morphological identification of both large (>0.5 mm in length) and small (<0.5 mm in length) invertebrate specimens intercepted at the Australian border. DNA barcode sequencing results supported the morphological identification in most cases and enabled immature stages of invertebrates and their eggs to be identified more confidently. Results also showed that sequencing the COI barcode region using the ONT rapid sequencing principle is a cost-effective and field-adaptable approach for the rapid and accurate identification of invertebrate pests. Overall, the results suggest that MinION sequencing of DNA barcodes offers a complementary tool to the existing morphological diagnostic approaches and provides rapid, accurate, reliable and defendable evidence for identifying invertebrate pests at the border.

Microbial Toxins in Insect and Nematode Pest Biocontrol

Invertebrate pests, such as insects and nematodes, not only cause or transmit human and livestock diseases but also impose serious crop losses by direct injury as well as vectoring pathogenic microbes. The damage is global but greater in developing countries, where human health and food security are more at risk. Although synthetic pesticides have been in use, biological control measures offer advantages via their biodegradability, environmental safety and precise targeting. This is amply demonstrated by the successful and widespread use of Bacillusthuringiensis to control mosquitos and many plant pests, the latter by the transgenic expression of insecticidal proteins from B. thuringiensis in crop plants. Here, I discuss the prospects of using bacterial and fungal toxins for pest control, including the molecular basis of their biocidal activity.

Phototoxicity of Ultraviolet-A against the Whitefly Bemisia tabaci and Its Compatibility with an Entomopathogenic Fungus and Whitefly Parasitoid

Ultraviolet (UV) radiation significantly affects insect life and, as a result, has been widely used to control different invertebrate pests. The current results demonstrate that when Bemisia tabaci first instar nymphs are exposed to UV-A light for 12, 24, 48, and 72 h, their developmental and biological parameters are negatively affected by UV-A exposure; the effect increased with an increase in exposure time. We hypothesized that UV-A light is compatible with other biological control agents. Results showed that when the entomopathogenic fungus Cordyceps fumosorosea was applied to third instar nymphs of B. tabaci previously exposed to UV-A light, the LC50 was 3.4% lower after 72 h of exposure to UV-A light compared to the control. However, when the fungus was exposed to UV-A light, its virulence decreased with an increase in UV-A exposure time. The parasitism rate of Encarsia formosa against 24 h UV-A-exposed third instar nymphs of B. tabaci increased while the adult emergence from parasitized nymphs was not affected after UV-A light exposure. Parasitism rate was significantly reduced however following E. formosa exposure to UV-A light; but again, adult emergence was not affected from parasitized nymphs. The percentage mortality of E. formosa increased with increasing exposure time to UV-A light. The enzyme activity of SOD, CAT, GST, and AChE and the energy reserve contents were negatively affected due to UV-A exposure. Collectively, this study has demonstrated that UV-A light significantly suppresses the immune system of B. tabaci and that UV-A light is compatible with other biological control agents if it is applied separately from the biological agent.

Management to Promote Flowering Understoreys Benefits Natural Enemy Diversity, Aphid Suppression and Income in an Agroforestry System

Agroforestry systems, where productive trees are integrated into agricultural land, can deliver benefits to biodiversity, natural pest control, and pollination, but the effects are highly variable. Recent advances in our understanding of flower strips in agricultural systems suggest that the management of the tree row understorey could be an important contributor to this variation. Here, we compare two cutting regimes for an understorey, originally seeded with the same flower mix, in the tree rows of an apple-arable agroforestry system: (i) uncut vegetation to promote a flowering understorey, and (ii) regularly mown vegetation. We recorded the effects of management on invertebrate pests, natural enemies, and pollinators, in both the apple and arable components. Apple trees above flowering understoreys supported significantly: (i) more natural enemies early in the season, (ii) fewer aphid colonies, (iii) fewer aphid-damaged fruits, and (iv) higher pollinator visitation, compared with those above mown understoreys. In the arable crop alleys, both the taxonomic richness and Shannon diversity of ground-based natural enemies were significantly higher adjacent to flowering understoreys, compared with those adjacent to mown understoreys, early in the season. Financial modelling based on aphid damage to apples, mowing costs, and income from Countryside Stewardship grants, indicated that flowering understoreys increased farm income by GBP 231.02 per ha of agroforestry compared with mown understoreys. Our results provide the first empirical evidence that management to promote flowering understoreys in agroforestry systems can be a win-win option to improve invertebrate diversity, associated ecosystem services, and farm income.

Phylogenomics of Tick Inward Rectifier Potassium Channels and Their Potential as Targets to Innovate Control Technologies

This study was conducted to enhance the identification of novel targets to develop acaricides that can be used to advance integrated tick-borne disease management. Drivers for the emergence and re-emergence of tick-borne diseases affecting humans, livestock, and other domestic animals in many parts of the world include the increased abundance and expanded geographic distribution of tick species that vector pathogens. The evolution of resistance to acaricides among some of the most important tick vector species highlights the vulnerability of relying on chemical treatments for tick control to mitigate the health burden of tick-borne diseases. The involvement of inward rectifier potassium (Kir) channels in homeostasis, diuresis, and salivary gland secretion in ticks and other pests identified them as attractive targets to develop novel acaricides. However, few studies exist on the molecular characteristics of Kir channels in ticks. This bioinformatic analysis described Kir channels in 20 species of hard and soft ticks. Summarizing relevant investigations on Kir channel function in invertebrate pests allowed the phylogenomic study of this class of ion channels in ticks. How this information can be adapted to innovate tick control technologies is discussed.