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.

Effects of pyrethroid and organophosphate insecticides on reared black soldier fly larvae (Hermetia illucens).

Black soldier fly larvae (Hermetia illucens) receive growing interest as a potential alternative animal feed source. These insects may be exposed to insecticide residues in the rearing substrate. This study aimed to investigate the effects of six different pyrethroid and organophosphate insecticides on this insect species' performance. The toxicity of two "model" substances for each of these classes (cypermethrin; pirimiphos-methyl) was quantified, with and without the synergist piperonyl butoxide (PBO). Critical effect doses corresponding to 10% yield (CED10) for cypermethrin (0.4 mg/kg) and pirimiphos-methyl (4.8 mg/kg) were determined. The addition of PBO to cypermethrin enhanced its relative potency with a factor 2.6. These data were compared against the relative toxicity of two analogue substances in each class (permethrin, deltamethrin; chlorpyrifos-methyl, malathion). Results suggest that exposure to concentrations complying with legal limits can cause significant reductions in yield. Exposure to multiple substances at lower concentrations resulted in negative additive and synergistic effects. Of the tested substances, deltamethrin was most toxic, causing 94% yield at 0.5 mg/kg. Analytical results suggested that transfer of tested substances to the larval biomass was substance- and concentration-specific, but appeared to be correlated to reduced yields and the presence of PBO. Transfer of organophosphates was overall low (<2%), but ranged from 8% to 75% for pyrethroids. Due to very low limits in insect biomass (∼0.01 mg/kg), high transfer may result in noncompliance. It is recommended that rearing companies implement lower contractual thresholds, and that policymakers consider adjusting legally allowed maximum residue levels in insect feed.

The use of animal by-products in a circular bioeconomy: Time for a TSE road map 3?

In 2005 and 2010, the European Commission (EC) published two subsequent 'Road Maps' to provide options for relaxation of the bans on the application of animal proteins in feed. Since then, the food production system has changed considerably and demands for more sustainability and circularity are growing louder. Many relaxations envisioned in the second Road Map have by now been implemented, such as the use of processed animal proteins (PAPs) from poultry in pig feed and vice versa. However, some legislative changes, in particular concerning insects, had not been foreseen. In this article, we present a new vision on legislation for increased and improved use of animal by-products. Six current legislative principles are discussed for the bans on animal by-products as feed ingredients: feed bans; categorization of farmed animals; prohibition unless explicitly approved; approved processing techniques, the categorization of animal by-products, and monitoring methods. We provide a proposal for new guiding principles and future directions, and several concrete options for further relaxations. We argue that biological nature of farmed animals in terms of dietary preferences should be better recognised, that legal zero-tolerance limits should be expanded if safe, and that legislation should be revised and simplified.

Aflatoxin B1 Metabolism of Reared Alphitobius diaperinus in Different Life-Stages.

The presence of carcinogenic aflatoxins in food and feed is a major issue. In prior studies, aflatoxin B1 (AfB1) and known primary metabolites were absent from Lesser Mealworm (LMW, Alphitobius diaperinus) reared on contaminated diets. LMW is a promising alternative protein source. The objectives of this stu\dy were to determine whether LMW can be reared on AfB1-contaminated feed in each life-stage, and to gather more insight into potential metabolites formed. Results suggested no adverse effects in terms of survival/growth when three stages of LMW (larvae, pre-pupae, beetles) were exposed to feed containing AfB1 concentrations of 200 and 600 µg/kg for 48 h. Insect and frass samples were analyzed by LC-MS/MS and high-resolution MS to, respectively, quantify concentrations of AfB1 and its major metabolites, and determine secondary metabolites. No AfB1 or major metabolites were quantified in the insect samples. Mass balance calculations showed that up to 40% of spiked AfB1 could be recovered in the frass, in the form of AfB1, aflatoxicol and AfM1. HRMS results suggested the presence of additional metabolites in the frass, but, due to lack of commercially available reference standards for these compounds, exact identification and quantification was not possible. More research is needed to verify the absence of toxicity.

Overcoming Technical and Market Barriers to Enable Sustainable Large-Scale Production and Consumption of Insect Proteins in Europe: A SUSINCHAIN Perspective.

The expected global population growth to 9.7 billion people in 2050 and the significant change in global dietary patterns require an increase in global food production by about 60%. The protein supply for feed and food is most critical and requires an extension in protein sources. Edible insects can upgrade low-grade side streams of food production into high-quality protein, amino acids and vitamins in a very efficient way. Insects are considered to be the "missing link" in the food chain of a circular and sustainable economy. Insects and insect-derived products have entered the European market since first being acknowledged as a valuable protein source for feed and food production in around 2010. However, today, scaling up the insect value chain in Europe is progressing at a relatively slow pace. The mission of SUSINCHAIN (SUStainable INsect CHAIN)-a four-year project which has received funding from the European Commission-is to contribute to novel protein provision for feed and food in Europe by overcoming the remaining barriers for increasing the economic viability of the insect value chain and opening markets by combining forces in a comprehensive multi-actor consortium. The overall project objective is to test, pilot and demonstrate recently developed technologies, products and processes, to realize a shift up to Technology Readiness Level 6 or higher. In addition to these crucial activities, the project engages with stakeholders in the insect protein supply chain for feed and food by living labs and workshops. These actions provide the necessary knowledge and data for actors in the insect value chain to decrease the cost price of insect products, process insects more efficiently and market insect protein applications in animal feed and regular human diets that are safe and sustainable. This paves the way for further upscaling and commercialization of the European insect sector.

Effects of insecticides on mortality, growth and bioaccumulation in black soldier fly (Hermetia illucens) larvae.

Residues of persistent insecticides may be present in the substrates on which insects are reared for food and feed, which may affect insect growth or survival. In addition, insecticidal substances may bio-accumulate in reared insects. The objective of this study was to assess potential effects of selected insecticides on the growth and survival of black soldier fly larvae (BSFL, Hermetia illucens) and on their safety when used as animal feed. Six insecticides (chlorpyrifos, propoxur, cypermethrin, imidacloprid, spinosad, tebufenozide) with different modes of action were tested in two sequential experiments. Cypermethrin was also tested with the synergist piperonyl butoxide (PBO). Standard BSFL substrate was spiked to the respective maximum residue level (MRL) of each insecticide allowed by the European Union to occur in feed; and BSFL were reared on these substrates. Depending on the observed effects in the first experiment, spiked concentrations tested in the second experiment were increased or reduced. At the concentrations applied (1 and 10 times MRL), three of the six tested substances (chlorpyrifos, propoxur, tebufenozide) did not affect the survival or biomass growth of BSFL, compared to the control (non-spiked) treatments. At MRL, imidacloprid stimulated the growth of BSFL compared to the controls. Spinosad and cypermethrin at the MRL level negatively affected growth and survival. The effects of cypermethrin appeared to be augmented by addition of PBO. A mean bio-accumulation factor of ≤0.01 was found in both experiments for all substances-except for cypermethrin, which was comparatively high, but still below 1 (0.79 at 0.1 mg/kg). The lack of accumulation of insecticides in the larvae suggests that there is no risk of larval products being uncompliant with feed MRLs. However, we conclude that insecticides present in substrates may affect growth and survival of BSFL. More research on a larger variety of substances and insect species is recommended.

The aflatoxin situation in Africa: Systematic literature review.

Contamination of African staple foods is a major issue for human and animal health, nutrition, and trade. This review aimed to collect and synthesize the available evidence on geographical spread, scale of contamination, disease burden, economic impact, and mitigation measures for aflatoxins in Africa by way of a systematic literature review. This knowledge can enhance management strategies for the major challenges to combat aflatoxins. The search was conducted by applying a predefined search strategy, using bibliographic databases and websites, covering the period 2010 to 2018. Results showed that maize, peanuts, and animal feeds were the most studied commodities. For maize, all studies indicated mean AFB1 to exceed the European Union legal limit. From studies on contamination levels and biomarkers, it is clear that overall exposure is high, leading to a substantial increase in long-term disease burden. In addition, concentrations in food occasionally can reach very high levels, causing acute aflatoxicoses. The trade-related impact of aflatoxin contamination was mainly evaluated from the standpoint of aflatoxin regulation affecting products imported from Africa. There was a limited number of studies on health-related economic impacts, pointing out a gap in peer-reviewed literature. A number of mitigation measures have been developed, but proof of cost-effectiveness or even costs alone of the practices is often lacking. We recommend more emphasis to be put in peer-reviewed studies on evidence-based cost-effective mitigation strategies for aflatoxins, on the scale and spread of the problem and its impacts on public health and economics for use in evidence-based policies.

Aflatoxin B1 Conversion by Black Soldier Fly (Hermetia illucens) Larval Enzyme Extracts.

The larvae of the black soldier fly (Hermetia illucens L., BSFL) have received increased industrial interest as a novel protein source for food and feed. Previous research has found that insects, including BSFL, are capable of metabolically converting aflatoxin B1 (AFB1), but recovery of total AFB1 is less than 20% when accounting for its conversion to most known metabolites. The aim of this study was to examine the conversion of AFB1 by S9 extracts of BSFL reared on substrates with or without AFB1. Liver S9 of Aroclor-induced rats was used as a reference. To investigate whether cytochrome P450 enzymes are involved in the conversion of AFB1, the inhibitor piperonyl butoxide (PBO) was tested in a number of treatments. The results showed that approximately 60% of AFB1 was converted to aflatoxicol and aflatoxin P1. The remaining 40% of AFB1 was not converted. Cytochrome P450s were indeed responsible for metabolic conversion of AFB1 into AFP1, and a cytoplasmic reductase was most likely responsible for conversion of AFB1 into aflatoxicol.

Bridging legal requirements and analytical methods: a review of monitoring opportunities of animal proteins in feed.

Availability and safety of food ranks among the basic requirements for human beings. The importance of the food producing sector, inclusive of feed manufacturing, demands a high level of regulation and control. This paper will present and discuss the relationships in the triangle of legislation, the background of hazards with a biological nature, and opportunities for monitoring methods, most notable for prion-based diseases as primary issue. The European Union legislation for prevention of prion-based diseases since 2000 is presented and discussed. The definitions and circumscriptions of groups of species will be analysed in the view biological classification and evolutionary relationships. The state of the art of monitoring methods is presented and discussed. Methods based on visual markers (microscopy), DNA-based methods (PCR), protein-based methods (ELISA, mass spectroscopy, proteomics), near infrared oriented methods and combinations thereof are being evaluated. It is argued that the use in legislation of non-homogeneous groups of species in a biological sense will hamper the optimal design of monitoring methods. Proper definitions are considered to act as bridges between legal demands and suitable analytical methods for effective monitoring. Definitions including specified groups of species instead of single species are more effective for monitoring in a range of cases. Besides the desire of precise circumscription of animal groups targeted by legislation, processed products need well defined definitions as well. Most notable examples are blood versus blood products, and hydrolysis of several types of material. The WISE principle for harmonising the design of legislation and of analytical methods is discussed. This principle includes the elements Witful (reasonable legal principles), Indicative (clear limits between prohibition and authorisation), Societal demands (public health, environment, economy), and Enforceable (presence of suited monitoring methods) in order to promote a balanced effort for reaching the desired level of safety in the food production chain.