Nosemosis is a microsporidian disease causing mortality and weakening of honey bee colonies, especially in the event of co-exposure to other sources of stress. As a result, the disease is regulated in some countries. Reliable and harmonised diagnosis is crucial to ensure the quality of surveillance and research results. For this reason, the first European Interlaboratory Comparison (ILC) was organised in 2017 in order to assess both the methods and the results obtained by National Reference Laboratories (NRLs) in counting Nosema spp. spores by microscopy. Implementing their own routine conditions of analysis, the 23 participants were asked to perform an assay on a panel of ten positive and negative samples of crushed honey bee abdomens. They were asked to report results from a qualitative and quantitative standpoint. The assessment covered specificity, sensitivity, trueness and precision. Quantitative results were analysed in compliance with international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). Three results showed a lack of precision and five a lack of trueness. However, overall results indicated a global specificity of 98% and a global sensitivity of 100%, thus demonstrating the advanced performance of the microscopic methods applied to Nosema spores by the NRLs. Therefore, the study concluded that using microscopy to detect and quantify spores of Nosema spp. was reliable and valid.

Vespa velutina nigrithorax is an invasive species of hornet accidentally introduced into Europe in 2004. It feeds on invertebrates, including honey bees, and represents a threat to European apiculture. In 2016, the first nest of this hornet was detected and destroyed on mainland UK. A further 8 nests were discovered between 2016 and 2019. Nest dissection was performed on all nests together with microsatellite analyses of different life stages found in the nests to address the reproductive output and success of nests found in the UK. None of the nests had produced the next generation of queens. Follow-up monitoring in those regions detected no new nests in the following years. Diploid males were found in many UK nests, while microsatellite analysis showed that nests had low genetic diversity and the majority of queens had mated with one or two males. All UK nests derived from the European zone of secondary colonisation, rather than from the native range of the species. None of the nests discovered so far have been direct offspring of another UK nest. The evidence suggests that these nests were separate incursions from a continental population rather than belonging to a single established UK population of this pest.

Waste biomass from the palm oil industry is currently burned as a means of disposal and solutions are required to reduce the environmental impact. Whilst some waste biomass can be recycled to provide green energy such as biogas, this investigation aimed to optimise experimental conditions for recycling palm waste into substrate for insects, farmed as a sustainable high-protein animal feed. NMR spectroscopy and LC-HRMS were used to analyse the composition of palm empty fruit bunches (EFB) under experimental conditions optimised to produce nutritious substrate rather than biogas. Statistical pattern recognition techniques were used to investigate differences in composition for various combinations of pre-processing and anaerobic digestion (AD) methods. Pre-processing methods included steaming, pressure cooking, composting, microwaving, and breaking down the EFB using ionic liquids. AD conditions which were modified in combination with pre-processing methods were ratios of EFB:digestate and pH. Results show that the selection of pre-processing method affects the breakdown of the palm waste and subsequently the substrate composition and biogas production. Although large-scale insect feeding trials will be required to determine nutritional content, we found that conditions can be optimised to recycle palm waste for the production of substrate for insect rearing. Pre-processing EFB using ionic liquid before AD at pH6 with a 2:1 digestate:EFB ratio were found to be the best combination of experimental conditions.

The productivity ofMusca domestica larvae reared on poultry litter and the nutritional parameters of insect meal as an animal feed ingredient have been evaluated. In laboratory bioassays, a seeding density of 0.32 g eggs/kg wet weight litter produced maximal yields of >50 g of naturally egressing larvae/kg substrate. Substrate composition and moisture content were critical determinants for insect development and generally litter wetted to water holding capacity (approx. 23% dry weight) was optimal for larval production. Four different processing methods were evaluated, but had no significant effects upon the nutritional quality of insect meals and all were effective in reducing microbial (Enterobacteriacae) populations to levels considered acceptable by feed regulations. Processed samples contained an average of 59.9% protein and 24.2% fat. Differences in levels of fatty acids, particularly palmitic, linoleic and α-linolenic were observed in larvae reared on different substrates. By contrast, highly similar amino acid profiles were derived from larvae reared on different substrates. With well-balanced amino acid profiles, superior to soymeal and more comparable to fishmeal,M. domestica larval meals have considerable potential for use as a protein-rich animal feed.

While the idea of using insect based feeds (IBFs) offers great potential, especially in developing countries, the environmental impact of implementation remains poorly researched. This study investigates the environmental performance of IBF production in the geographical context of West Africa. Drawing on published life cycle inventory (LCIs) data, the impact of three different IBF production systems were ex-ante evaluated (ReCiPe method) and compared to conventional feed resources. The explorative life cycle study provides a basis for trade-off analysis between different insect rearing systems (Musca domestica and Hermetia illucens) and provides insights on the environmental performance of IBF in comparison with conventional animal- and plant based protein feeds (fishmeal, cottonseed and soybean meal). The impacts of IBFs were shown to be largely determined by rearing techniques and the environmental loads of rearing substrates, attesting advantages to the rearing of housefly (M. domestica) larvae on chicken manure and the use of natural oviposition, i.e., substrate inoculation through naturally occurring flies. A comparison with conventional feeds pointed out the environmental disadvantages of current IBF production designs (especially in comparison to plant based feeds) that were largely attributable to their different position in the trophic network (decomposers) and the systems' sub-standard capacity utilisation (insufficient economy of scale effect). When larvae are reared on substrates of low economic value (i.e., waste streams), IBF impacts were comparable to fishmeal. The results of the comparative assessment also highlighted a methodological limitation in the ReCiPe method, which does not account for impacts related to the use of biotic resources. As a consequence, the utilization of naturally grown resources, such as wild anchoveta, was treated as an ecosystem service of no environmental charge, providing disproportionate advantages to the fishmeal system.

The red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is a significant pest affecting a wide variety of different stored products around the Globe. Despite its economic impact, there is evidence that the lures currently used in traps to monitor for this species are largely ineffective. Based on the evolutionary history of T. castaneum, and the ecological niche it occupies, the volatiles of wheat germ oil and volatiles produced by grain-associated fungi have the potential to act as attractants for this species. We used electroantennography (EAG) to measure the electrophysiological response elicited by sixty-eight volatile compounds found in wheat germ oil and/or grain-associated fungi in two T. castaneum strains; an established lab population (CTC12 strain) and a recently caught wild population. Many volatile compounds from both sources elicited strong antennal depolarisations, and the responses of both strains were highly correlated. We then tested whether the compounds that triggered the strongest antennal depolarisations also elicited behavioural responses by using Y-tube olfactometer bioassays and identified several compounds attractive to both strains. The discovery of novel compounds that elicit strong EAG signals and behavioural responses could prove useful in the design of improved lures for T. castaneum and other stored product pests. Our future research will identify how effective these attractive volatiles might be when used in combination, and when used under conditions that more closely replicate a stored product environment.

The yellow-legged or Asian hornet (Vespa velutina colour form nigrithorax) was introduced into France from China over a decade ago. Vespa velutina has since spread rapidly across Europe, facilitated by suitable climatic conditions and the ability of a single nest to disperse many mated queens over a large area. Yellow-legged hornets are a major concern because of the potential impact they have on populations of many beneficial pollinators, most notably the western honey bee (Apis mellifera), which shows no effective defensive behaviours against this exotic predator. Here, we present the first report of this species in Great Britain. Actively foraging hornets were detected at two locations, the first around a single nest in Gloucestershire, and the second a single hornet trapped 54 km away in Somerset. The foraging activity observed in Gloucestershire was largely restricted to within 700 m of a single nest, suggesting highly localised movements. Genetic analyses of individuals from the Gloucestershire nest and the single hornet from Somerset suggest that these incursions represent an expansion of the European population, rather than a second incursion from Asia. The founding queen of the Gloucestershire nest mated with a single male, suggesting that sexual reproduction may have occurred in an area of low nest density. Whilst the nest contained diploid adult males, haploid ‘true’ males were only present at the egg stage, indicating that the nest was detected and removed before the production of queens. Members of the public reported additional dead hornets associated with camping equipment recently returned from France and imported timber products, highlighting possible pathways of incursion. The utility of microsatellites to inform surveillance during an incursion and the challenge of achieving eradication of this damaging pest are discussed.

While the concept of insect based feeds (IBFs) promises great potential, especially in developing countries, the sustainability performance of IBF production remains widely underexplored. Drawing on experimental data from rearing trials in West Africa, three different insect production systems were modelled ex-ante. The generic models served as a basis to analyse and compare the process performances of different IBF production systems using Musca domestica and Hermetia illucens reared on different substrates. The results show that the input efficiency in the production of IBF is largely determined by the quality of rearing substrates, the larval development time and the employed inoculation practises, i.e., the method by which eggs or larvae are added to rearing substrates. The H. illucens system ranked highest for conversion efficiency (substrate input per IBF output), but showed substantially higher inputs in labour, fossil energy and output of wastewater. M. domestica systems operated at lower conversion efficiencies, which resulted in higher outputs of residue substrates, together with higher emissions, land requirements, built infrastructure and water. By offering full disclosure of generic inventory data, this study provides data and inspiration for prospect research and development activities and offers a reference to future life cycle assessments (LCAs) on IBF.

Due to the rapid growth of the world's population and the increasing demand for food, there is an urgent need for alternative, more sustainable sources of protein. Insects have an important role in the diet in some societies and current initiatives are exploring the potential that insects have to offer for the production of food and feed. In this context, the safety implications of both producing and consuming insects are an important aspect to investigate. Here we present a bioinformatics analysis of proteomics data obtained for larvae of four different species of fly to assess the homology of tropomyosin, arginine kinase and myosin light chain with the crustacean orthologous proteins and other known allergenic proteins. The results indicate that the three proteins share homology with known allergens and therefore it is likely that they are also potential allergens. The implications in relation to mass rearing of flies are discussed.

There is an urgent need to increase the supply of sustainable protein for use in animal feed and the use of insect protein provides a potential alternative to protein crops and fishmeal. For example, fly larvae are highly compatible with use in animal feed containing much digestible protein with levels of key amino acids that are comparable with those found in high value alternatives such as soybean. However, the safety of protein from insects and subsequently the meat and fish from animals fed on such a diet requires further assessment. Here we present safety data from the larvae of the four fly species that have perhaps the greatest economic relevance in relation to their use as animal feed being: house fly (Musca domestica), blue bottle (Calliphora vomitoria), blow fly (Chrysomya spp.) and black soldier fly (Hermetia illucens). Diverse rearing methods were used to produce larvae fed on a range of waste substrates and in four geographically dispersed locations being; UK, China, Mali and Ghana. Chemical safety data were collected by a fully accredited laboratory in the UK. The levels of the main subclasses of chemical contaminants considered for animal feed were determined, being; veterinary medicines, pesticides, heavy metals, dioxins and polychlorinated biphenyls, polyaromatic hydrocarbons and mycotoxins. The larvae analysed generally possessed levels of chemical contaminants which were below recommended maximum concentrations suggested by bodies such as the European Commission, the World Health Organisation and Codex. However, the toxic heavy metal cadmium was found to be of concern in three of the M. domestica samples analysed.

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The small hive beetle Aethina tumida is an endemic parasitic pest and scavenger of colonies of social bees indigenous to sub-Saharan Africa. In this region this species rarely inflicts severe damage on strong colonies since the bees have developed strategies to combat them. However, A. tumida has since ‘escaped’ from its native home and has recently invaded areas such as North America and Australia where its economic impact on the apiculture industry has been significant. Small hive beetle, should it become established within Europe, represents a real and live threat to the UK bee keeping industry. Here we review the biology and current pest status of A. tumida and up to-date research in terms of both chemical and biological control used against this honey bee pest.

Adult female Eulophus pennicornis require a source of nutrition, provided by sources such as pollen, nectar and honeydew or by host feeding, to promote longevity and facilitate egg production. There is potential for parasitoids to be exposed directly to contaminants, including gene products in transgenic crops, through feeding on plant materials, honeydew or hosts. Among such potential contaminants are lectins such as Galanthus nivalis agglutinin (GNA) and concanavalin agglutinin (Con A). The effect of direct exposure to honey diets containing GNA and Con A on the longevity and fecundity of E. pennicornis was examined. These lectins have been expressed in a number of plant species for the control for various insect pests. Both GNA and Con A significantly reduced longevity and fecundity at the highest concentration used (0.5% w/v). The effect on fecundity was shown to be related to a reduction in longevity. Examination of the gustatory response of adult female E. pennicornis to honey diet containing 1% w/v GNA or Con A revealed no significant differences in consumption rate on first exposure. A significant reduction in the time spent feeding on diet containing 1% Con A was found on the second exposure to the diet. This could have been the result of either a conditioned aversion response or the intoxication of the insect. The effect of Con A on longevity and fecundity could have been, in part, a result of reduced food intake. Studies on nutrition and egg resorption demonstrated that the availability of honey solution prolongs the longevity of E. pennicornis and the lack of a source of nutrition promotes oosorption. A greater understanding of feeding behaviour and ovigeny is required to understand fully the potential ecological consequence of transgenic crops on parasitoid species through routes of direct exposure to transgene products.

Chemical insecticides are used to protect stored grain from infestation by stored product insects and mites. In the UK only a limited number of products are available for application and there are concerns about safety, pest resistance and environmental impact of these conventional pesticides. Biological control offers an alternative to the use of chemical insecticides. The potential for biological control of storage pests in the UK using an insect-specific fungus, Beauveria bassiana , to treat the structure of the stores, has previously been established. However, this study also highlighted areas where improvements were needed; specifically to improve the uptake of the fungal conidia by the pests and to improve their germination and penetration into the pests. In addition it was necessary to ensure that potential formulations had a good shelf-life and to develop a mass production method to consistently produce high quality fungal conidia. A four year project has recently been completed examining these  areas in detail. The work has concentrated on two different fungal isolates of B. bassiana , both of which were found from insects in UK grain stores. Optimisation of production methods, formulation and delivery systems has resulted in prototype formulations that exhibit good viability over periods up to one year and that have good efficacy against a range of storage insect pests under conditions that are likely to be found in UK grain stores. Pilot scale trials using three species of stored product beetle have shown that significant levels of control can be achieved. An overview of the key findings is presented. The study has made a significant contribution to the development of a biopesticide as a structural treatment for grain storage areas in the UK. Keywords : Biological control; Beauveria bassiana; Oryzaephilus surinamensis ; Structural treatment; Biopesticide

Fusion proteins have considerable potential as novel insect control agents because they enable the oral delivery of insecticidal peptides to the haemolymph of pests. Transport is achieved via fusion of the toxin to a carrier protein Galanthus nivalis agglutinin (GNA) that, after ingestion, binds to and crosses the insect gut epithelia. A fusion protein comprising a toxin from the South Indian red scorpion (Mesobuthus tamulus) that is fused to a GNA polypeptide (ButaIT/GNA) has a detrimental effect on the development of tomato moth Lacanobia oleracea (L.) (Lepidoptera: Noctuidae) larvae. The present study examines the effects of ButaIT/GNA and GNA, delivered orally or by injection, on the development of L. oleracea larvae, and the subsequent effects on the gregarious ectoparasitoid Eulophus pennicornis (Nees) (Hymenoptera: Eulophidae) developing on ButaIT/GNA- and GNA-treated hosts. The fusion protein, but not GNA, reduces the growth of fifth stadium L. oleracea larvae. The development of E. pennicornis is not affected by the presence of ButaIT/GNA in hosts that ingest the protein, although it is affected when hosts are injected with the protein. This difference is considered to be a result of higher levels of fusion protein being present when the fusion protein is injected. Intact ButaIT/GNA is detected by immunoassay in the haemolymph of L. oleracea larvae after ingestion of the fusion protein. More unexpectedly, negative effects are observed for the growth of E. pennicornis larvae developing on hosts that have either ingested, or been injected with GNA.

Coccinella septempunctata is known to actively avoid substrates treated with an insecticide containing the organophosphate dimethoate. This study examines the responses of C. septempunctata to a range of different insecticide products from three chemical classes, carbamates, organophosphates and pyrethroids. Five formulated product insecticides were compared with the active ingredients (AIs) at two different application rates and using two different spray patterns (conferring choice or no-choice designs) in a randomised block design. Coccinellids' responses differed between insecticide classes. Pyrethoid treatments significantly reduced locomotion. Organophosphates effected mixed locomotory responses, as found in previous studies. Carbamate treatments effected very few changes in locomotory activity. Similar results were found at both application rates tested and under different test designs. The results of the AI tests indicated that different components of the products were responsible for the different reactions, with the AIs being responsible for some responses but the carriers being responsible for others. Results are discussed in relation to the insecticides' modes of action and to their potential to increase the efficiency of integrated aphid control.

"Maintaining Aethina tumida (Coleoptera: Nitidulidae) under quarantine laboratory conditions in the UK and preliminary observations on its behaviour." Journal of Apicultural Research, 47(3), pp. 192–193

Commodities may be attacked by a multitude of pest species simultaneously and so to be cost-effective any hire must be attractive to a range of target species. The objective of this study was to test a multi-species lure formulation with different dispensers under conditions as close as possible to their use in practice. The attractant effect of the multi-species lure formulation was tested using two types of dispensers with populations of the three principal grain beetle pests in the UK (Oryzaephilus surinamensis, Sitophilus granarius and Cryptolestes ferrugineus). The trials were performed over a period of six weeks with the lures tested in PC(TM) Traps: first in a grain bulk and in six grain bins and second in the surrounding area using PC(TM) Floor Traps. The amounts of volatiles released in and around ill situ traps were measured using Solid-Phase Microextraction (SPME).The first lure dispenser tested was attractive to O. surinamensis and C ferrugineus but it attracted fewer S. granarius than the control traps. It was shown that most of the volatiles were released at the beginning of the trial and therefore lures were not effective over the whole six-week period. The second lure dispenser released the attractant volatiles more consistently over six weeks and exerted significant. attraction to O. surinamensis and C ferrugineus in PC(TM) Traps in the grain bulk and to O. surinamensis and S. granarius in floor traps. Too few C ferrugineus were caught in floor traps to make a comparison. The reason for the repellent effect on S. granarius in the grain bulk was unclear and this is discussed along with the importance of measuring volatiles released by the lure. Crown Copyright (C) 2008 Published by Elsevier Ltd. All rights reserved.