Mentha spp. essential oils: toxicity to Alphitobius diaperinus, activity against poultry pathogenic bacteria, and Beauveria bassiana compatibility.

The botanical insecticide market is growing because of limitations placed on the use of certain synthetic chemical insecticides. In this sense, the lesser mealworm Alphitobius diaperius (Coleoptera: Tenebrionidae) is the main poultry pest. The insect causes weight loss and damage to the digestive system of poultry, and it is a vector and reservoir of pathogens. Consequently, this study explored the following hypotheses: (i) essential oils (EOs) derived from Mentha spp. are toxic to A. diaperius; (ii) these EOs are compatible with Beauveria bassiana, the natural enemy of the poultry pest, that parasite A. diaperinus; (iii) these EOs also exhibit activity against bacteria that are pathogenic to poultry. In topical applications and ingestion tests, EOs from Mentha arvensis, Mentha spicata, and Mentha piperita were toxic to A. diaperinus. Chromatographic analyses revealed that menthol is the predominant compound in M. arvensis and M. piperita, whereas carvone is the major compound in M. spicata. Both (-)- and (+)-menthol, along with (-)- and (+)-carvone, underwent testing with A. diaperinus. Nevertheless, their activity was not as potent as those of the EOs, suggesting a possible synergistic and/or additive effect. The EOs did not have any adverse effects on the conidial germination, vegetative growth, or conidia production per colony of the entomopathogenic fungus B. bassiana. Consequently, these EOs are compatible with this natural enemy. The EO extracted from M. spicata exhibited significant toxicity against Staphylococcus aureus (ATCC 25923), whereas the remaining EOs displayed moderate toxicity against this bacterium. The EOs derived from Mentha spp., as assessed in this study, hold promise for the development of botanical insecticides tailored for the control of A. diaperinus. These insecticides are selective in favor of the natural enemy B. bassiana and can also serve as effective sanitizers, thanks to their antibacterial properties.

Effects of dietary exposure to plant toxins on bioaccumulation, survival, and growth of black soldier fly (Hermetia illucens) larvae and lesser mealworm (Alphitobius diaperinus).

In their natural habitat, insects may bioaccumulate toxins from plants for defence against predators. When insects are accidently raised on feed that is contaminated with toxins from co-harvested herbs, this may pose a health risk when used for human or animal consumption. Plant toxins of particular relevance are the pyrrolizidine alkaloids (PAs), which are genotoxic carcinogens produced by a wide variety of plant species and the tropane alkaloids (TAs) which are produced by a number of Solanaceae species. This study aimed to investigate the transfer of these plant toxins from substrates to black soldier fly larvae (BSFL) and lesser mealworm (LMW). PAs and the TAs atropine and scopolamine were added to insect substrate simulating the presence of different PA- or TA-containing herbs, and BSFL and LMW were grown on these substrates. Bioaccumulation from substrate to insects varied widely among the different plant toxins. Highest bioaccumulation was observed for the PAs europine, rinderine and echinatine. For most PAs and for atropine and scopolamine, bioaccumulation was very low. In the substrate, PA N-oxides were quickly converted to the corresponding tertiary amines. More research is needed to verify the findings of this study at larger scale, and to determine the potential role of the insect and/or substrate microbiome in metabolizing these toxins.

Five Surfaces Treated with d-Tetramethrin plus Acetamiprid for the Management of Tenebrio molitor and Alphitobius diaperinus: Which Is the Best?

Tenebrio molitor L. (Coleoptera: Tenebrionidae) and Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) are two common tenebrionids occurring in grain storages. In this study, we assessed the immediate and delayed mortalities caused by d-tetramethrin plus acetamiprid on five different surfaces, i.e., plastic, glass, metal, wood, and ceramic, against adults of the two species. The tests included two label doses of the insecticide (minimum and maximum) and two food scenarios (food and no food). Generally, the maximum dose was more efficient than the minimum dose, and the presence of food resulted in lower observed mortalities than when food was absent. Tenebrio molitor was more susceptible than A. diaperinus, at all dose, food, and surface scenarios. At delayed bioassays, both doses killed all T. molitor on plastic, while on wood, mortality ranged between 80.6 and 100.0%, regardless of the food scenario. Concerning A. diaperinus, delayed mortalities ranged among treated surfaces, food scenarios, and dose from 58.3 to 100.0%. The insecticide killed the most individuals when it was treated on glass, while when it was applied on wood caused the death of the least individuals. Concerning plastic, metal, and ceramic surfaces, no general trend was observed. The maximum dose of the tested insecticide provides elevated mortalities for both species when food is absent.

Population Growth of Three Stored Product Beetle Species on Alphitobius diaperinus (Coleoptera: Tenebrionidae) Meals.

The lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), is an important pest of stored products and at the same time a species with great potential as food and feed that has recently attracted increasing interest as nutrient source. Future predictions show a massive increase of insect meals' production in the near future, therefore, as expected in the case of other durable stored products, insect meals may get infested by insects during their storage. In this context and as a continuance of our previous study on the susceptibility of meal of the yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae), to storage insect infestations, the objective of this study was to test the susceptibility of A. diaperinus meals to infestations of three stored products pests, namely A. diaperinus itself, T. molitor, and the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). The population growth of the three species was evaluated on pure A. diaperinus meal, as well as on A. diaperinus meal-based substrates with different percentages of wheat bran (0, 25, 50, 90, and 100% bran). Our results showed that all three insect species tested were able to grow and develop on the A. diaperinus meal-based substrates evaluated, giving in a short time high population densities. This study confirms again our initial theory for insect infestations during storage of insect-based products.

Challenges in the Biological Control of Pests in Poultry Production: a Critical Review of Advances in Brazil.

Poultry farming is an important activity in animal protein production worldwide, either by laying hen farming or broilers. Over the last decades, the change in production systems with confinement of large numbers of hens has increased productivity and reduced costs; however, it has also increased sanitary issues. In this setting, arthropods that are adapted to poultry houses have gained great importance. They cause direct damage to hens, either by blood spoliation caused by ectoparasites or lesions in the digestive tract (e.g., lesser mealworm) or by indirect damages, by transmitting pathogens or by affecting egg quality, when they attain pest status. Synthetic chemical products comprise the most frequently used control strategy against these pests, with relative efficacy and many side effects. In Europe, some countries also adopt alternative prevention or control measures. In Brazil, however, although there are some groups of researchers that work on developing alternative control, its use is virtually zero. The present review shows a critical overview of this context in Brazil, based on the alternatives that have already been studied and made available, but have not been implemented, yet, and the potential stumbling blocks created by the very poultry market against these advances.

Safety of frozen and freeze-dried formulations of the lesser mealworm (Alphitobius diaperinus larva) as a Novel food pursuant to Regulation (EU) 2015/2283.

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on frozen and dried formulations from whole lesser mealworm (Alphitobius diaperinus larva) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The term lesser mealworm refers to the larval form of the insect species Alphitobius diaperinus. The NF comprises the frozen and freeze-dried formulations of the lesser mealworm as whole or in the form of a paste or powder. Apart from water in the frozen formulations (whole, paste), the main components of the NF are crude protein and fat, besides smaller amounts of digestible carbohydrates and fibre (chitin). The Panel notes that the levels of contaminants in the NF depend on the concentration of such substances in the insect feed. The Panel notes furthermore that the true protein levels in the NF are overestimated when using the nitrogen-to-protein conversion factor of 6.25, due to the presence of non-protein nitrogen from chitin. The applicant proposed to use the NF formulations added as an ingredient to various food products such as cereal bars, pasta, meat imitates and bakery products. The target population is the general population. Additionally, the applicant proposed to use the NF as a food supplement in adults. The Panel notes that, considering that the NF will not be the sole source of dietary protein, and the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted subchronic 90-day toxicity study with the NF as testing material did not raise safety concerns. The Panel considers that the consumption of the NF may induce primary sensitisation and allergic reactions to lesser mealworm proteins and may cause allergic reactions in subjects with allergy to crustaceans and dust mites. Additionally, allergens from the feed may end up in the NF. Allergenicity aside, the Panel concludes that the NF is safe under the proposed uses and use levels.

Efficacy of a Biocidal Paint in Controlling Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) and Improving the Quality of Air and Litter in Poultry Houses.

Effective disinfection and disinsection are the keys to successful operation of modern poultry farms and the safety of poultry products. The cleaning and disinfection of poultry houses are important aspects of farm hygiene management. The correct execution of all steps of cleaning, disinfection, and disinsection procedures and the use of appropriate products are crucial for the prevention and control of zoonotic and animal diseases. In this study it was assumed that a water-based slow-release biocidal paint could be useful in controlling insect pests such as Alphitobius diaperinus and reducing microbiological contamination of air and litter in poultry houses and have a beneficial effect on microclimate in poultry houses. Therefore, the locations of A. diaperinus in the poultry houses, the microbiological contamination of air and litter, as well as the microclimatic conditions in the houses and the physicochemical parameters of the litter were evaluated. The results suggest that the tested biocidal paint could be an effective alternative to other insecticides and disinfectants. Additionally, the research is of a practical nature and may be very useful for poultry producers in controlling A. diaperinus populations and maintaining proper hygiene in poultry houses. Further research is needed.

Short- and Long-Term Mortalities of Small and Large Larvae of Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) on Concrete Surfaces Treated with Three Insecticides: Impact of Food.

The lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) is an important stored-product pest for the poultry industry as it is a vector of dangerous pathogens for humans. In the present study, we evaluated the short- and long-term mortalities of small and large larvae of A. diaperinus when they were exposed to concrete-covered Petri dishes treated with etofenprox, deltamethrin, and the combination of piperonyl butoxide+acetamiprid+d-tetramethrin. Small and large larvae were exposed to each insecticide applied on concrete surfaces with or without food. The short-term mortality was recorded after 1 day, 3 days, 5 days, and 7 days, while the long-term mortality was recorded 7 days after the transport of the larvae alive to pesticide-free concrete-covered dishes. Regarding short-term mortality levels, 97.8% and 80.0% of the small and large larvae, that were exposed to etofenprox without food, died after 7 days of exposure, respectively. Concerning deltamethrin, all tested small larvae were killed after 3 days (without food) and 5 days (with food) of exposure. For large larvae, deltamethrin caused 98.9% (with food) and 100.0% (without food) mortality levels after 5 days of exposure. The combination of piperonyl butoxide+acetamiprid+d-tetramethrin caused high mortality levels to small larvae, i.e., 84.4% and 100.0% on dishes with and without food, respectively, but low to moderate mortality levels to large larvae that did not exceed 67.8% after 7 days of exposure. Long-term mortality varied vastly among the tested insecticides. Etofenprox killed 100.0% of the small larvae on concrete without food, but 24.0% of the large larvae exposed to concrete containing food. Deltamethrin did not provide long-term mortality to large larvae when food was present. Piperonyl butoxide+acetamiprid+d-tetramethrin caused mortality rates that overall varied from 33.5% (large larvae on concrete with food) to 85.0% (small larvae on concrete with food). In conclusion, deltamethrin killed almost all exposed larvae at exposures of ≤5 days, regardless of their size and the presence of food on the concrete.

Detection of Alphitobius diaperinus by Real-Time Polymerase Chain Reaction With a Single-Copy Gene Target.

Use of edible insects as an alternative source of proteins in food and feed is increasing. These last years, numerous companies in Europe have started producing insects for food and feed purposes. In the European Union, the use of edible insects for human consumption falls within Regulation (EU) No. 2015/2283 on novel foods. For feed, Commission Regulation (EU) 2017/893 authorizes seven insect species as processed animal proteins for aquaculture. Methods of authentication are required to check the conformity of the products. In this study, we propose a real-time polymerase chain reaction (PCR) method for the specific detection of the lesser mealworm (Alphitobius diaperinus), one of the species included in the shortlist of authorized insects. The selected target is the cadherin gene with a single-copy (per haploid genome) illustrated by our experimental evidence. The PCR test amplified a 134-bp fragment of the cadherin gene. The qualitative method was assessed toward several performance criteria. Specificity was checked against 54 insect species next to other animal and plant species. The sensitivity, efficiency, robustness, and transferability of the PCR assay were also successfully tested. Finally, the applicability of the test was assessed on real-life processed samples (industrial meals) of A. diaperinus. The study also showed that there seems to be a huge confusion on the correct labeling of the marketed mealworms. We did not succeed to get Alphitobius laevigatus samples. They all appeared to belong to the A. diaperinus taxon.

Polyphenol Loaded W1/O/W2 Emulsions Stabilized with Lesser Mealworm (Alphitobius diaperinus) Protein Concentrate Produced by Membrane Emulsification: Stability under Simulated Storage, Process, and Digestion Conditions.

Water-in-oil-in-water (W1/O/W2) emulsions are complex delivery systems for polyphenols amongst other bio-actives. To stabilize the oil-water interphase, dairy proteins are commonly employed, which are ideally replaced by other, more sustainable sources, such as insect proteins. In this study, lesser mealworm (Alphitobius diaperinus) protein concentrate (LMPC) is assessed and compared to whey protein (WPI) and pea protein (PPI), to stabilize W1/O/W2 emulsions and encapsulate a commercial polyphenol. The results show that LMPC is able to stabilize W1/O/W2 emulsions comparably to whey protein and pea protein when using a low-energy membrane emulsification system. The final droplet size (d4,3) is 7.4 µm and encapsulation efficiency is between 72 and 74%, regardless of the protein used. Under acidic conditions, the LMPC shows a similar performance to whey protein and outperforms pea protein. Under alkaline conditions, the three proteins perform similarly, while the LMPC-stabilized emulsions are less able to withstand osmotic pressure differences. The LMPC stabilized emulsions are also more prone to droplet coalescence after a freeze-thaw cycle than the WPI-stabilized ones, but they are the most stable when exposed to the highest temperatures tested (90 °C). The results show LMPC's ability to stabilize multiple emulsions and encapsulate a polyphenol, which opens the door for application in foods.

Effect of cypermethrin on the control of lesser mealworm (Alphitobius diaperinus) and broiler performance / Efeito da cipermetrina no controle de cascudinho (Alphitobius diaperinus) sobre o desempenho de frangos de corte

This study analysed the effectiveness of using different levels of a product based on 6% cypermethrin and 2% citronella on broiler production and control the lesser mealworm (Alphitobius diaperinus). A total of 648 one-day-old chicks (Cobb®) which were allocated to one of three treatments (no application of the commercial product - control, and two concentrations of the product 2.00 and 3.33g/m2) in eight replicates with 27 birds each. The commercial product, which was based on cypermethrin was applied in the chambers with an atomizer. The lesser mealworm population was sampled weekly (1, 7, 14, 21, 28, 35 and 42 days of the experiment) using traps, at three points (front, middle and back) inside the pen. The performance, carcass and cut yields were evaluated. At 21, 28, 35 and 42 days of the experiment, the number of lesser mealworm adults and larvae was lower in the treatment groups involving 2.00 and 3.33g/m2 of the product than in the control group. On the first and 14th days of the study, the number of lesser mealworms adults was lower in the group treated with 3.33g/m2 of the product compared to the control group, although the results did not differ statistically from those obtained with 2.00g/m2 of the product. In conclusion, the application of the commercial product at both concentrations (2.00 and 3.33g/m2) was effective in combating Alphitobius diaperinus adults and larvae, and the performance was not influenced by treatments.(AU)

Este estudo analisou a eficácia do uso de diferentes níveis de um produto à base de cipermetrina a 6% e citronela a 2%, na produção de frangos de corte e no controle do cascudinho (Alphitobius diaperinus). Um total de 648 pintos de um dia de idade (Cobb®) foram distribuídos em três câmaras com os tratamentos (sem aplicação do produto comercial - controle, e duas concentrações do produto comercial - 2,00 e 3,33g/m2) em oito repetições com 27 aves cada. O produto comercial a base de cipermetrina foi aplicado nas câmaras com um atomizador. A população de cascudinho foi amostrada semanalmente (1, 7, 14, 21, 28, 35 e 42 dias do experimento) usando armadilhas, em três pontos (frente, meio e fundo) de cada parcela. Foram avaliados o desempenho, rendimentos de carcaça e cortes. Aos 21, 28, 35 e 42 dias do experimento, o número de adultos e larvas de cascudinho foi menor nos grupos de tratamento envolvendo 2,00 e 3,33g/m2 do produto quando comparado ao grupo controle. No primeiro e no 14º dia do estudo, o número de cascudinhos adultos foi menor no grupo tratado com 3,33g/m2 do produto em comparação ao grupo controle, embora os resultados não tenham diferido estatisticamente daqueles obtidos com 2,00g/m2 do produto. A conversão alimentar de 1 a 21 dias de idade foi significativa, com os melhores resultados para essa variável quando aplicado 3,33g do produto por metro quadrado e no tratamento controle. Em conclusão, a aplicação do produto comercial em ambas as concentrações (2,00 e 3,33g/m2) foi eficaz no combate a adultos e larvas de Alphitobius diaperinus e, não influenciou o desempenho.(AU)

Effect of cypermethrin on the control of lesser mealworm (Alphitobius diaperinus) and broiler performance

ABSTRACT: This study analysed the effectiveness of using different levels of a product based on 6% cypermethrin and 2% citronella on broiler production and control the lesser mealworm (Alphitobius diaperinus). A total of 648 one-day-old chicks (Cobb®) which were allocated to one of three treatments (no application of the commercial product - control, and two concentrations of the product 2.00 and 3.33g/m2) in eight replicates with 27 birds each. The commercial product, which was based on cypermethrin was applied in the chambers with an atomizer. The lesser mealworm population was sampled weekly (1, 7, 14, 21, 28, 35 and 42 days of the experiment) using traps, at three points (front, middle and back) inside the pen. The performance, carcass and cut yields were evaluated. At 21, 28, 35 and 42 days of the experiment, the number of lesser mealworm adults and larvae was lower in the treatment groups involving 2.00 and 3.33g/m2 of the product than in the control group. On the first and 14th days of the study, the number of lesser mealworms adults was lower in the group treated with 3.33g/m2 of the product compared to the control group, although the results did not differ statistically from those obtained with 2.00g/m2 of the product. In conclusion, the application of the commercial product at both concentrations (2.00 and 3.33g/m2) was effective in combating Alphitobius diaperinus adults and larvae, and the performance was not influenced by treatments.

RESUMO: Este estudo analisou a eficácia do uso de diferentes níveis de um produto à base de cipermetrina a 6% e citronela a 2%, na produção de frangos de corte e no controle do cascudinho (Alphitobius diaperinus). Um total de 648 pintos de um dia de idade (Cobb®) foram distribuídos em três câmaras com os tratamentos (sem aplicação do produto comercial - controle, e duas concentrações do produto comercial - 2,00 e 3,33g/m2) em oito repetições com 27 aves cada. O produto comercial a base de cipermetrina foi aplicado nas câmaras com um atomizador. A população de cascudinho foi amostrada semanalmente (1, 7, 14, 21, 28, 35 e 42 dias do experimento) usando armadilhas, em três pontos (frente, meio e fundo) de cada parcela. Foram avaliados o desempenho, rendimentos de carcaça e cortes. Aos 21, 28, 35 e 42 dias do experimento, o número de adultos e larvas de cascudinho foi menor nos grupos de tratamento envolvendo 2,00 e 3,33g/m2 do produto quando comparado ao grupo controle. No primeiro e no 14º dia do estudo, o número de cascudinhos adultos foi menor no grupo tratado com 3,33g/m2 do produto em comparação ao grupo controle, embora os resultados não tenham diferido estatisticamente daqueles obtidos com 2,00g/m2 do produto. A conversão alimentar de 1 a 21 dias de idade foi significativa, com os melhores resultados para essa variável quando aplicado 3,33g do produto por metro quadrado e no tratamento controle. Em conclusão, a aplicação do produto comercial em ambas as concentrações (2,00 e 3,33g/m2) foi eficaz no combate a adultos e larvas de Alphitobius diaperinus e, não influenciou o desempenho.

Effectiveness of Biocidal Paint Containing Permethrin, Ultramarine and Violet 23 Against Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) in Laboratories and Poultry Houses.

Reducing Alphitobius diaperinus in poultry production is a difficult task. However, attempts should be made to control the insect pest, as it poses a serious threat to the life and health of the chickens, as well as the workers on a farm. Our research was conducted in two stages to assess the effectiveness of the biocidal paint against A. diaperinus, containing active substances such as permethrin and a mixture of ultramarine and violet 23. In the first stage, under laboratory conditions, after 22 days, 100% mortality of A. diaperinus larvae and adults was achieved. This allowed us to assume that the biocidal paint may also be effective in poultry houses. In the poultry house where biocidal paint was applied, the number of insects decreased continuously alongside the sampling dates. In both research stages, the biocidal paint proved more effective against A. diaperinus than traditional limewash, and also the time to effective interaction of the paint was noted. Additionally, it was observed that the larvae were more susceptible to the active substances than adults. The research was practical, however, further analyses are necessary to fully control A. diaperinus, especially in poultry houses.

Use of Starch Granules Enriched with Carvacrol for the Lesser Mealworm, Alphitobius diaperinus Control in Chicken House: Effects on Insects and Poultry.

The aim of this study was to investigate the effect of starch granules enriched with carvacrol and mixed with straw pellets (as poultry litter) on the mortality of larvae and adults of the lesser mealworm, Alphitobius diaperinus Panzer, a cosmopolitan pest inhabiting chicken houses in vast numbers worldwide. Additionally, the effect of starch granules on the growth parameters and survival of broiler chickens exposed to treated litter was examined. In this study, granules containing 3, 5, and 10% carvacrol was used. In a simulated chicken house bioassay, this material was mixed with pellets in three different proportions: 30/70%, 40/60%, and 50/50% (granules/pellets, respectively). On this medium, young larvae (approximately 10 days old), older larvae (last stage before pupa), and unsexed 7-10 days old adults of the lesser mealworm, with access to food, were colonized. Experiments were performed at 29°C in the dark. The study shows that poultry litter with the addition of starch granules enriched with 10% of carvacrol in the proportion of 40:60% (granules:pellets) appears to be the optimal medium applicable to broiler houses for A. diaperinus control. In this environment, all larvae and adults died within 3-4 days and the overall development of the experimental chickens was similar to that of the control. However, the feed conversion rate was slightly higher in the treated group (1.72) than in the control group (1.56). The average final body weight in the treated group was 100 g lower than that in the control group (the differences were not statistically significant).

Agri-Food Side-Stream Inclusion in The Diet of Alphitobius Diaperinus. Part 2: Impact on Larvae Composition.

Insects are gaining interest as an alternative protein source for feed/food purposes. Although the lesser mealworm (LM) is commercially produced for human consumption, published data on its nutrient composition is scarce. This study reports on LM larvae reared on 18 different diets composed of side-streams to (1) determine the nutritional composition of the larvae and (2) study the effect of dietary changes on the larval nutrient composition. The LM larvae proved to be of good nutritional value with essential amino acids profiles comparable with that of beef and linoleic acid (C18:2) was the most dominant essential fatty acids in the larvae. The side-stream based diets varied on dry matter basis in protein (16-34%) and lipid content (2-19%). The nutrient content of the larvae reared on diets that supported good growth ranged between 37% and 49% of protein, 22% and 26% of lipid and 4% to 6% of chitin on dry matter basis. No significant correlations were identified between the larval protein or lipid content and that of the diet, but it was found between the diet nutrients and larval growth. Based on larval growth data and economic considerations, diets composed of wheat middlings with a 10-15% inclusion of rapeseed meal were identified as suitable feed for LM. Highest larval yields were obtained with diets containing 15-22% of proteins and 5-10% of lipids.

Frozen storage of lesser mealworm larvae (Alphitobius diaperinus) changes chemical properties and functionalities of the derived ingredients.

The effect of frozen storage on the chemical properties and ingredient functionalities of Lesser mealworms was investigated at -20 °C for 2 months. Major changes occurred in the first week of frozen storage. Proteins, among which heavy chain myosin, underwent denaturation and aggregation, as shown by a decrease in solubility, SDS-PAGE pattern, and Confocal Laser Scanning Microscopy. The ice melting point in larvae was -32.5 °C as determined by DSC: 25% of water is not frozen at -20 °C, possibly due to anti-freezing proteins preventing ice formation. The presence of unfrozen water favoured various enzymatic activities as shown by a pH decrease, indicating protein hydrolysis. The molecular changes during frozen storage increased the browning reactions due to phenoloxidase activity. Foaming ability, foam stability and gel network stability increased upon frozen storage due to protein denaturation. Results provide important information regarding the opportunity of frozen storage of insect larvae for both research and industrial purposes.

Agri-Food Side-Stream Inclusions in the Diet of Alphitobius diaperinus Part 1: Impact on Larvae Growth Performance Parameters.

Insects are attracting increased attention in western countries as a protein source for feed and food industries. Currently, insect farmers use high-quality (cereal-based) diets. Part of the ingredients in these diets can also be used directly in food applications. To avoid competition and improve the sustainable and economical aspect of insect rearing, a search for alternative insect diets is ongoing. Side-streams from the agri-food sector offer potential. The lesser mealworm (Alphitobius diaperinus) is an insect that is commercially reared on large scale for food application. The current paper reports on six agri-food side-streams that were included in the diet of the lesser mealworm. The impact of 29 diets (single side-streams or mixtures) on the larvae growth was evaluated by monitoring the larval yield, efficiency of conversion of ingested feed, and larval weight. The larvae were able to grow on all diets, but differences in growth were observed. Two side-streams, wheat middlings and rapeseed meal, were proven to support good larval performance when used as a single ingredient. A combination of these two with brewery grains as moisture source provided (1) the best larval growth and (2) the most economically profitable diet. In conclusion, this study illustrates successful rearing of the lesser mealworm on side-stream-based diets.

Effect of endogenous phenoloxidase on protein solubility and digestibility after processing of Tenebrio molitor, Alphitobius diaperinus and Hermetia illucens.

Upon extracting soluble proteins from insects as potential food ingredient, endogenous enzymes, such as phenoloxidases, are expected to negatively affect protein properties. The effect of phenoloxidases on solubility and digestibility of proteins was investigated for larvae of Tenebrio molitor, Alphitobius diaperinus and Hermetia illucens. Phenoloxidase inhibition was done using blanching (50 s, 90 °C) before extraction or extracting in presence of sulfite. Similar soluble protein yields and compositions were found without and with sulfite addition, whereas blanching decreased soluble protein yield. Upon in-vitro hydrolysis by pepsin and trypsin, soluble proteins from H. illucens were more digestible than those of T. molitor and A. diaperinus. Phenoloxidase activity during grinding negatively affected in-vitro pepsin hydrolysis. Besides phenoloxidase activity, also endogenous proteases were shown to remain active at pH 8 in extracts containing sulfite and after blanching of larvae. This stresses that protease activity needs to be carefully controlled in the design of insect based ingredients.

Commercially Available Essential Oil Formulas as Repellents Against the Stored-Product Pest Alphitobius diaperinus.

The main aim of the presented paper is to assess the potential repellent effect of selected essential oils (EOs) against the lesser mealworm (Alphitobius diaperinus), which can cause economic losses in storage and in the poultry industry. Due to the development of pesticide resistance in A. diaperinus populations, as well as an attempt to limit extensive use of potentially harmful pesticides in food-related industries, there is a strong need for the development of alternative methods of dealing with A. diaperinus infestations. Because of their cost-effectiveness, availability and low vertebrate toxicity, EOs are promising agents in pest management. In the presented paper four off-the-shelf EOs: mint, vanilla, lemon and citronella (and mixtures of them) were tested as potential repellents. Moreover, a novel preference assay, providing an extended analysis of the preference and the locomotor response, was used. The most effective EOs were: citronella and lemon. EOs mixtures were generally more repellent than individual EOs, with the lemon and vanilla 1:1 mixture acting as the strongest repellent. A few of the tested EOs caused significant alterations to the locomotor activity, although no direct relation was observed. In conclusion, EOs can be potentially used as repellent agents in A. diaperinus management. Additionally, data on the locomotor activity may lead to designing better push-pull strategies in pest management.

Susceptibly of Alphitobius diaperinus to Beauveria bassiana extracts.

Alphitobius diaperinus is an important pathogen with worldwide distribution that causes severe economic loss of efficiency in broilers. This study evaluates the potential of organic extracts of two strains entomopathogenic fungus Beauveria bassiana (CG71 and UNI40) as a biocontrol agent on A. diaperinus and promotes the phytochemical investigation. The effective percentages of mortalities were 95.97% (UNI40 methanolic extract), 69.23, 64.64, and 50.39% (CG 71 methanolic, ethyl acetate and butanol extracts). However, there was a decrease in the lesser mealworms susceptibility in relation to the use of insecticides and extracts. The metabolites 5-hydroxymethyl-2-furanoic acid, dipicolinic acid and monomethyl dipicolinate were isolated from ethyl acetate extract, and ß-adenosine of butanolic extract of B. bassiana CG 71. In addition, the cyclodepsipeptides were identified in methanolic extracts of the two strains. The insecticide activity results indicated that the B. bassiana extracts are an alternative to A. diaperinus control.