An international inter-laboratory study on Nosema spp. spore detection and quantification through microscopic examination of crushed honey bee abdomens.

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.

Managing incursions of Vespa velutina nigrithorax in the UK: an emerging threat to apiculture.

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.

From waste to food: Optimising the breakdown of oil palm waste to provide substrate for insects farmed as animal feed.

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 invasion, provenance and diversity of Vespa velutina Lepeletier (Hymenoptera: Vespidae) in Great Britain.

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.

The physical action of three diatomaceous earths against the cuticle of the flour mite Acarus siro L. (Acari: Acaridae).

Experiments examined the accepted modes of action of the desiccant dust, diatomaceous earth (DE), against the flour mite, Acarus siro (L.) (Astigmata). Adult mites were exposed to three DE products for periods of 3, 18 and 72 h under conditions designed to allow partial desiccation of the mites without causing mortality. After exposure, the DE dust particles were washed off the mite bodies, and both the DE and the mites were examined for presence of cuticular hydrocarbons by gas chromatography-mass spectroscopy (GC-MS) analysis. GC-MS identified seven major cuticular lipids of chain length C(13)--C(26) that may have a role in the waterproofing of A. siro. After 18 h exposure, n-tridecane and several different long-chain fatty acid ethyl esters were detected on one of the DE products. After 72 h, n-tridecane was detected on all three DE products. Mite samples retained after removal of the DEs were examined by microscopy. Tentative evidence was observed by conventional low-power microscopy that might indicate uniform removal of the epicuticle. However, a detailed examination by scanning electron microscopy showed no signs of abrasion.

Identification of a sex pheromone from male yellow mealworm beetles, Tenebrio molitor.

The sex pheromone released by the adult female Tenebrio molitor, 4-methyl-1-nonanol, is well known. In addition, there is evidence that adult males release a pheromone that attracts females. The purpose of the present study was to isolate and identify male-released pheromone(s). Emissions from virgin adult males and females were collected on filter paper and extracted with pentane. Extracts were analyzed by gas chromatography-mass spectrometry. One male-specific compound was detected and identified as (Z)-3-dodecenyl acetate (Z3-12:Ac). In arena bioassays, E3-12:Ac was attractive to females only, at 1 and 10 microg doses. E3-12:Ac was also attractive to females at a 10-microg dose. The presence of both male and female pheromones, each attracting the opposite sex, may contribute to maintaining a high-density population of both sexes.

Effectiveness of the BT mite trap for detecting the storage mite pests, Acarus siro, Lepidoglyphus destructor and Tyrophagus longior.

Traps have been used extensively to provide early warning of hidden pest infestations. To date, however, there is only one type of trap on the market in the U.K. for storage mites, namely the BT mite trap, or monitor. Laboratory studies have shown that under the test conditions (20 degrees C, 65% RH) the BT trap is effective at detecting mites for at least 10 days for all three species tested: Lepidoglyphus destructor, Tyrophagus longior and Acarus siro. Further tests showed that all three species reached a trap at a distance of approximately 80 cm in a 24 h period. In experiments using 100 mites of each species, and regardless of either temperature (15 or 20 degrees C) or relative humidity (65 or 80% RH), the most abundant species in the traps was T. longior, followed by A. siro then L. destructor. Trap catches were highest at 20 degrees C and 65% RH. Temperature had a greater effect on mite numbers than humidity. Tests using different densities of each mite species showed that the number of L. destructor found in/on the trap was significantly reduced when either of the other two species was dominant. It would appear that there is an interaction between L. destructor and the other two mite species which affects relative numbers found within the trap.