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The use of light-emitting diode (LED) lights in horticulture allows growers to adjust the light spectrum to optimize crop production and quality. However, changes in light quality can also influence plant–arthropod interactions, with possible consequences for pest management. The addition of far-red light has been shown to interfere with plant immunity, thereby increasing plant susceptibility to biotic stress and increasing pest performance. Far-red light also influences plant emission of volatile organic compounds (VOCs) and might thus influence tritrophic interactions with biological control agents. We investigated how far-red light influences the VOC-mediated attraction of the predatory mite Phytoseiulus persimilis to tomato plants infested with Tetranychus urticae, and its ability to control T. urticae populations. RESULTS: Far-red light significantly influences herbivore-induced VOC emissions of tomato plants, characterized by a change in relative abundance of terpenoids, but this did not influence the attraction of P. persimilis to herbivore-induced plants. Supplemental far-red light led to an increased population growth of T. urticae and increased numbers of P. persimilis. This resulted in a stronger suppression of T. urticae populations under supplemental far-red light, to similar T. urticae numbers as in control conditions without supplemental far-red light. CONCLUSION: We conclude that supplemental far-red light can change herbivore-induced VOC emissions but does not interfere with the attraction of the predator P. persimilis. Moreover, far-red light stimulates biological control of spider mites in glasshouse tomatoes due to increased population build-up of the biocontrol agent.

Reliable options to control the cabbage root fly, Delia radicum L., are lacking in many countries as restrictions on insecticide use have tightened due to environmental concerns. Although microbial control agents are often considered as a sustainable alternative, their application in agriculture is constrained by inconsistent efficacy owing to low field persistence. To stimulate naturally occurring beneficial microbes, soil amendment with the residual streams of insect production has been suggested as an alternative to synthetic fertilization and a new approach to microbial crop protection. In a set of greenhouse experiments, exuviae and frass of black soldier fly larvae, Hermetia illucens L., house crickets, Acheta domesticus L. and exuviae of mealworms, Tenebrio molitor L., were added to soil from an organically managed field. Exuviae and frass treatments were compared to treatments with synthetic fertilizer. Brussels sprouts, Brassica oleracea L., plants were grown in amended soil for 5 weeks before being infested with cabbage root fly larvae. Insect and plant performance were assessed by recording cabbage root fly survival, biomass and eclosion time and seed germination and plant biomass, respectively. Whereas soil amendment with black soldier fly frass or exuviae reduced cabbage root fly survival and biomass, respectively, amendment with house cricket or mealworm residual streams did not negatively affect root fly performance. Furthermore, seed germination was reduced in soil amended with house cricket exuviae, while amendment with either residual stream derived from black soldier fly larvae or house crickets resulted in lower plant shoot biomass compared with the synthetic fertilizer treatment. Amending soil with black soldier fly residual streams could become a novel and low-cost tool to be integrated in cabbage root fly management programmes, especially where methods currently available are insufficient. Therefore, the mechanisms underlying the effects of insect-derived soil amendments described here should be the focus of future research.

The house cricket, Acheta domesticus, is a commonly reared insect for food and feed purposes. In 1977, a report described a colony collapse, which was caused by the single-stranded DNA virus Acheta domesticus densovirus (AdDV). Currently, there are no confirmed A. domesticus colonies free of AdDV, and viral disease outbreaks are a continuous threat to A. domesticus mass rearing. Correlations between cricket rearing density or temperature and AdDV abundance have been hypothesized, but experimental evidence is lacking. Optimised rearing conditions, including temperature and density, are key to cost-effective cricket production. In this study, house crickets were subjected to different combinations of rearing density (10, 20, 40 crickets per box) and temperature (25, 30, 35 °C) to study the effect on cricket survival, biomass, and AdDV abundance. Rearing temperature affected had a minor effect on survival, which ranged between 80 and 83%. Total cricket biomass increased with higher temperatures and higher densities. Viral abundance in crickets at the end of the rearing period was variable; however, high rearing density seemed to result in higher AdDV abundance. At 35 °C, a temperature considered suboptimal for house cricket production, viral abundance tended to be lower than at 25 or 30 °C.

Climate change alters many environmental parameters with strong consequences for ecological interactions, from species interactions to community dynamics. Temperature is crucial in determining ecosystem dynamics, especially for those involving ectothermic species such as plants or insects. Phenotypic plasticity, the capacity of one genotype to produce different phenotypes in response to environmental conditions, is a common mechanism by which individuals adapt to changing environments and is observed in multiple traits. The capacity of genotypes to adapt to novel temperature conditions plays a crucial role in structuring ecosystem dynamics and species persistence in adverse conditions. It is well recognised that temperature in natural ecosystems fluctuates over multiple time scales (e.g., hour, day, season, year). These fluctuations can follow predictable patterns or be unpredictable, with different consequences for phenotypic plasticity and ecosystem dynamics. Among trophic interactions, host–parasitoid interactions represent a special case because of the intimate symbiosis of the parasitoid larvae with their host. Understanding how and to what extent phenotypic plasticity structures species’ ecological niches is of utmost importance in the context of rapid climate change. With a particular focus on host–parasitoid interactions, this review discusses the literature on the role of phenotypic plasticity in fluctuating environments, highlighting the role of temporal dynamics. While we discuss literature on phenotypic plasticity at large, this review emphasises the fundamental effects of extreme temperatures in driving biochemical rates underlying phenotypic plasticity.

Insect farming is a rapidly expanding agricultural sector with the potential to improve the environmental sustainability of livestock production. A holistic approach is needed to use farmed insects responsibly and effectively with broad benefits across food systems.

With climate change, the occurrence of extreme weather events such as heatwaves is expected to increase. Extreme temperature events are short periods of high temperature (i.e., >95th percentile of temperature distribution), and their occurrences are difficult to predict. These extreme temperature events pose great challenges to living organisms, especially ectotherms such as insects whose body temperature is greatly influenced by environmental temperature. Temperature is known to play an essential role in driving insect performance as well as inducing behavioral changes. Exposure to extreme temperatures causes physiological damage leading to potential long-lasting effects on insect performance. We aimed at assessing the impact of direct (i.e., current) and indirect (previously experienced) exposure to extreme temperature on the oviposition success of Diadegma semiclausum Hellén (Hymenoptera: Ichneumonidae). Diadegma semiclausum is a larval endoparasitoid of Plutella xylostella L. (Lepidoptera: Plutellidae), a worldwide pest of brassicaceous crops. Oviposition success, measured as the percentage of parasitized P. xylostella, was significantly lower at 33 °C than at 25 °C. However, previous exposure to 33 °C did not significantly impact the egg-laying performance of D. semiclausum, indicating that, after a heatwave, the egg-laying performance can recover. Neither the direct nor indirect exposure to extreme temperature altered D. semiclausum host-instar preference. These results indicate that temperature directly impacts D. semiclausum parasitism performance, although more detailed research is needed to disentangle direct physiological mechanisms from behavioral responses. Overall, these study results confirm others, suggesting that control of P. xylostella by D. semiclausum is likely to be impaired by more frequent heatwaves predicted under climate change scenarios.

Plants face attackers aboveground and belowground. Insect root herbivores can lead to severe crop losses, yet the underlying transcriptomic responses have rarely been studied. We studied the dynamics of the transcriptomic response of Brussels sprouts (Brassica oleracea var. gemmifera) primary roots to feeding damage by cabbage root fly larvae (Delia radicum), alone or in combination with aboveground herbivory by cabbage aphids (Brevicoryne brassicae) or diamondback moth caterpillars (Plutella xylostella). This was supplemented with analyses of phytohormones and the main classes of secondary metabolites; aromatic, indole and aliphatic glucosinolates. Root herbivory leads to major transcriptomic rearrangement that is modulated by aboveground feeding caterpillars, but not aphids, through priming soon after root feeding starts. The root herbivore downregulates aliphatic glucosinolates. Knocking out aliphatic glucosinolate biosynthesis with CRISPR-Cas9 results in enhanced performance of the specialist root herbivore, indicating that the herbivore downregulates an effective defence. This study advances our understanding of how plants cope with root herbivory and highlights several novel aspects of insect–plant interactions for future research. Further, our findings may help breeders develop a sustainable solution to a devastating root pest.

Plants interact with a diversity of phytophagous insects above- and belowground. By inducing plant defence, one insect herbivore species can antagonize or facilitate other herbivore species feeding on the same plant, even when they are separated in space and time. Through systemic plant-mediated interactions, leaf-chewing herbivores may affect the preference and performance of root-feeding herbivores. We studied how six different leaf-chewing herbivore species of Brassica oleracea plants affected oviposition preference and larval performance of the root-feeding specialist Delia radicum. We expected that female D. radicum flies would oviposit where larval performance was highest, in accordance with the preference–performance hypothesis. We also assessed how the different leaf-chewing herbivore species affected defence-related gene expression in leaves and primary roots of B. oleracea, both before and after infestation with the root herbivore. Our results show that leaf-chewing herbivores can negatively affect the performance of root-feeding D. radicum larvae, although the effects were relatively weak. Surprisingly, we found that adult D. radicum females show a strong preference to oviposit on plants infested with a leaf-chewing herbivore. Defence-related genes in primary roots of B. oleracea plants were affected by the leaf-chewing herbivores, but these changes were largely overridden upon local induction by D. radicum. Infestation by leaf herbivores makes plants more attractive for oviposition by D. radicum females, while decreasing larval performance. Therefore, our findings challenge the preference–performance hypothesis in situations where other herbivore species are present.

Root herbivores pose a major threat to agricultural crops. They are difficult to control and their damage often goes unnoticed until the larvae reach their most devastating late instar stages. Crop diversification can reduce pest pressure, generally without compromising yield. We studied how different diversified cropping systems affected the oviposition and abundance of the specialist cabbage root fly Delia radicum, the most important root herbivore in Brassica crops. The cropping systems included a monoculture, pixel cropping, and four variations of strip cropping with varying intra- and interspecific crop diversity, fertilization and spatial configuration. Furthermore, we assessed whether there was a link between D. radicum and other macroinvertebrates associated with the same plants. Cabbage root fly oviposition was higher in strip cropping designs compared to the monoculture and was highest in the most diversified strip cropping design. Despite the large number of eggs, there were no consistent differences in the number of larvae and pupae between the cropping systems, indicative of high mortality of D. radicum eggs and early instars especially in the strip cropping designs. D. radicum larval and pupal abundance positively correlated with soil-dwelling predators and detritivores and negatively correlated with other belowground herbivores. We found no correlations between the presence of aboveground insect herbivores and the number of D. radicum on the roots. Our findings indicate that root herbivore presence is determined by a complex interplay of many factors, spatial configuration of host plants, and other organisms residing near the roots.

Soil composition and herbivory are two environmental factors that can affect plant traits including flower traits, thus potentially affecting plant–pollinator interactions. Importantly, soil composition and herbivory may interact in these effects, with consequences for plant fitness. We assessed the main effects of aboveground insect herbivory and soil amendment with exuviae of three different insect species on visual and olfactory traits of Brassica nigra plants, including interactive effects. We combined various methodological approaches including gas chromatography/mass spectrometry, spectroscopy and machine learning to evaluate changes in flower morphology, colour and the emission of volatile organic compounds (VOCs). Soil amended with insect exuviae increased the total number of flowers per plant and VOC emission, whereas herbivory reduced petal area and VOC emission. Soil amendment and herbivory interacted in their effect on the floral reflectance spectrum of the base part of petals and the emission of 10 VOCs. These findings demonstrate the effects of insect exuviae as soil amendment on plant traits involved in reproduction, with a potential for enhanced reproductive success by increasing the strength of signals attracting pollinators and by mitigating the negative effects of herbivory.

The black soldier fly Hermetia illucens Linnaeus (Diptera: Stratiomyidae; BSF) is gaining interest as an alternative protein ingredient for livestock feed. Larval aggregation behaviour occurs commonly in larvae of various dipteran species. However, the cues initiating aggregation behaviour and its occurrence in subsequent larval instars are still unknown in BSF larvae. Here, we focus on understanding the attraction of larvae to cues left behind in the substrate by conspecific larvae. We developed a dual-choice test in the dark and examined the preference behaviour of different instars with the use of video recordings. Larval choices were identified based on first substrate contact, cumulative number of substrate contacts, entry of the substrate and number of larvae present in a substrate over time. Our results demonstrate that BSF larvae discriminated between similar substrates with or without cues released by conspecifics, with a strong preference for previously colonised substrates. Our results also showed a difference in behaviour between head-capsule classes. Substrate contacts occurred more frequently in larvae from a lower head-capsule class and the number of larvae present in a substrate over time differed between the head-capsule classes 0.71–0.80 mm and 0.91–1.00 mm. Demonstrating arrestment in response to chemical cues from conspecifics is the first step of understanding aggregation behaviour of BSF larvae and offers opportunities to identify the chemical cues involved.

Insect herbivores are amongst the most destructive plant pests, damaging both naturally occurring and domesticated plants. As sessile organisms, plants make use of structural and chemical barriers to counteract herbivores. However, over 75 percent of herbivorous insect species are well adapted to their host's defenses and these specialists are generally difficult to ward off. By actively antagonizing the number of insect eggs deposited on plants, future damage by the herbivore's offspring can be limited. Therefore, it is important to understand which plant traits influence attractiveness for oviposition, especially for specialist insects that are well adapted to their host plants. In this study, we investigated the oviposition preference of Pieris butterflies (Lepidoptera: Pieridae) by offering them the choice between 350 different naturally occurring Arabidopsis thaliana accessions. Using a genome-wide association study of the oviposition data and subsequent fine mapping with full genome sequences of 164 accessions, we identified WRKY42 and AOC1 as candidate genes that are associated with the oviposition preference observed for Pieris butterflies. Host plant choice assays with A. thaliana genotypes impaired in WRKY42 or AOC1 function confirmed a clear role for WRKY42 in oviposition preference of female Pieris butterflies, while for AOC1 the effect was mild. In contrast, WRKY42-impaired plants, that were preferred for oviposition by butterflies, negatively impacted offspring performance. These findings exemplify that plant genotype can have opposite effects on oviposition preference and caterpillar performance. This knowledge can be used for breeding trap crops or crops that are unattractive for oviposition by pest insects.

Background Novel malaria vector control approaches aim to combine tools for maximum protection. This study aimed to evaluate novel and re-evaluate existing putative repellent ‘push’ and attractive ‘pull’ components for manipulating the odour orientation of malaria vectors in the peri-domestic space. Methods Anopheles arabiensis outdoor human landing catches and trap comparisons were implemented in large semi-field systems to (i) test the efficacy of Citriodiol® or transfluthrin-treated fabric strips positioned in house eave gaps as push components for preventing bites; (ii) understand the efficacy of MB5-baited Suna-traps in attracting vectors in the presence of a human being; (iii) assess 2-butanone as a CO2 replacement for trapping; (iv) determine the protection provided by a full push-pull set up. The air concentrations of the chemical constituents of the push–pull set-up were quantified. Results Microencapsulated Citriodiol® eave strips did not provide outdoor protection against host-seeking An. arabiensis. Transfluthrin-treated strips reduced the odds of a mosquito landing on the human volunteer (OR 0.17; 95% CI 0.12–0.23). This impact was lower (OR 0.59; 95% CI 0.52–0.66) during the push-pull experiment, which was associated with low nighttime temperatures likely affecting the transfluthrin vaporisation. The MB5-baited Suna trap supplemented with CO2 attracted only a third of the released mosquitoes in the absence of a human being; however, with a human volunteer in the same system, the trap caught < 1% of all released mosquitoes. The volunteer consistently attracted over two-thirds of all mosquitoes released. This was the case in the absence (‘pull’ only) and in the presence of a spatial repellent (‘push-pull’), indicating that in its current configuration the tested ‘pull’ does not provide a valuable addition to a spatial repellent. The chemical 2-butanone was ineffective in replacing CO2. Transfluthrin was detectable in the air space but with a strong linear reduction in concentrations over 5 m from release. The MB5 constituent chemicals were only irregularly detected, potentially suggesting insufficient release and concentration in the air for attraction. Conclusion This step-by-step evaluation of the selected ‘push’ and ‘pull’ components led to a better understanding of their ability to affect host-seeking behaviours of the malaria vector An. arabiensis in the peri-domestic space and helps to gauge the impact such tools would have when used in the field for monitoring or control. Graphical Abstract

1. Plants interact with various organisms, aboveground as well as belowground. Such interactions result in changes in plant traits with consequences for members of the plant-associated community at different trophic levels. Research thus far focussed on interactions of plants with individual species. However, studying such interactions in a community context is needed to gain a better understanding. 2. Members of the aboveground insect community induce defences that systemically influence plant interactions with herbivorous as well as carnivorous insects. Plant roots are associated with a community of plant-growth promoting rhizobacteria (PGPR). This PGPR community modulates insect-induced defences of plants. Thus, PGPR and insects interact indirectly via plant-mediated interactions. 3. Such plant-mediated interactions between belowground PGPR and aboveground insects have usually been addressed unidirectionally from belowground to aboveground. Here, we take a bidirectional approach to these cross-compartment plant-mediated interactions. 4. Recent studies show that upon aboveground attack by insect herbivores, plants may recruit rhizobacteria that enhance plant defence against the attackers. This rearranging of the PGPR community in the rhizosphere has consequences for members of the aboveground insect community. This review focusses on the bidirectional nature of plant-mediated interactions between the PGPR and insect communities associated with plants, including (a) effects of beneficial rhizobacteria via modification of plant defence traits on insects and (b) effects of plant defence against insects on the PGPR community in the rhizosphere. We discuss how such knowledge can be used in the development of sustainable crop-protection strategies.

Insects such as the black soldier fly (BSF) are a nutritious feed component for livestock with high protein levels. BSF can be reared on a wide range of organic residual streams. This allows for local production within a circular agriculture, decoupling livestock production from import of expensive feed components, such as fishmeal or soymeal. Rearing of BSF can be done by smallholder farmers, thus contributing to their livelihood, economic sustainability and social status. Smallholder farmers contribute importantly to food security, which is a prerequisite for a stable society. In armed conflicts, smallholder farmers are usually the first to suffer. In countries recovering from conflict, agricultural development should focus on restoring food production by smallholder farmers, improving their socio-economic position, thereby contributing to sustainable development goals 2 (zero hunger) and 16 (peace and justice). Here, we focus on these SDGs with an example of reintegration of ex-combatants as smallholder insect producers in post-conflict Colombia.

Changing the spectral composition of light has numerous advantages for the production and quality of horticultural crops. However, changing the light spectrum to stimulate growth can have negative consequences for plant responses to biotic stress. Especially changes in the ratio between red and far-red light (R:FR) have an important effect on plant-herbivore interactions. A low R:FR is indicative of shading and competition for light and induces strong growth responses which are known as shade avoidance responses, including stem elongation and leaf hyponasty. Exposure to low R:FR also inhibits plant defensive responses against biotic agents and increases the performance of pests and pathogens. In this study we tested whether an increase in R:FR, through the supplementation of red light LEDs can be used to reduce the performance of arthropod herbivores. Tomato plants (Solanum lycopersocum) were exposed to three different R:FR ratios; 0.5 (shading), 1.2 (sunlight) and 5.2 (artificial) and infested with either caterpillars (Manduca sexta), spider mites (Tetranychus urticae), aphids (Myzus persicae) or whiteflies (Trialeurodes vaporariorum). Plants exposed to a low R:FR showed clear stem elongation and leaf hyponasty consistent with the shade-avoidance responses, while plants exposed to high R:FR showed reduced stem length and reduced hyponasty. The performance of all four herbivore species was significantly increased after exposure to low R:FR, although the strength of this effect varied between phloem feeding and tissue feeding herbivores. Increasing the R:FR only reduced the performance of the caterpillars and did not significantly affect the other herbivore species. These results indicate that herbivore species with different feeding strategies respond differently to changes in light quality. These findings have implications for the use of LEDs in greenhouse horticulture.

Saprophagous fly larvae interact with a rich community of bacteria in decomposing organic matter. Larvae of some species, such as the black soldier fly, can process a wide range of organic residual streams into edible insect biomass and thus produce protein as a sustainable component of livestock feed. The microbiological safety of the insects and substrates remains a point of concern. Substrate-associated bacteria can dominate the larval gut microbiota, but the larvae can also alter the bacterial community in the substrate. However, the relative importance of substrate type and larval density in bacterial community dynamics is unknown. We investigated four larval densities (0 [control], 50, 100, or 200 larvae per container [520 mL; diameter, 75 mm]) and three feed substrates (chicken feed, chicken manure, and camelina substrate [50% chicken feed, 50% camelina oilseed press cake]) and sampled the bacterial communities of the substrates and larvae at three time points over 15 days. Although feed substrate was the strongest driver of microbiota composition over time, larval density significantly altered the relative abundances of several common bacterial genera, including potential pathogens, in each substrate and in larvae fed chicken feed. Bacterial communities of the larvae and substrate differed to a higher degree in chicken manure and camelina than in chicken feed. This supports the substrate-dependent impact of black soldier fly larvae on bacteria both within the larvae and in the substrate. This study indicates that substrate composition and larval density can alter bacterial community composition and might be used to improve insect microbiological safety.

Plants interact with a diversity of phytophagous insects above- and belowground. By inducing plant defence, one insect herbivore species can antagonize or facilitate other herbivore species feeding on the same plant, even when they are separated in space and time. Through systemic plant-mediated interactions, leaf-chewing herbivores may affect the preference and performance of root-feeding herbivores. We studied how six different leaf-chewing herbivore species of Brassica oleracea plants affected oviposition preference and larval performance of the root-feeding specialist Delia radicum. We expected that female D. radicum flies would oviposit where larval performance was highest, in accordance with the preference-performance hypothesis. We also assessed how the different leaf-chewing herbivore species affected defence-related gene expression in leaves and primary roots of B. oleracea, both before and after infestation with the root herbivore. Our results show that leaf-chewing herbivores can negatively affect the performance of root-feeding D. radicum larvae, although the effects were relatively weak. Surprisingly, we found that adult D. radicum females show a strong preference to oviposit on plants infested with a leaf-chewing herbivore. Defence-related genes in primary roots of B. oleracea plants were affected by the leaf-chewing herbivores, but these changes were largely overridden upon local induction by D. radicum. Infestation by leaf herbivores makes plants more attractive for oviposition by D. radicum females, while decreasing larval performance. Therefore, our findings challenge the preference-performance hypothesis in situations where other herbivore species are present.

Background Insects have recently been identified as a more sustainable protein-dense food source and may represent a viable alternative to conventional animal-derived proteins. Objectives We aimed to compare the impacts of ingesting lesser mealworm– and milk-derived protein on protein digestion and amino acid absorption kinetics, postprandial skeletal muscle protein synthesis rates, and the incorporation of dietary protein–derived amino acids into de novo muscle protein at rest and during recovery from exercise in vivo in humans. Methods In this double-blind randomized controlled trial, 24 healthy, young men ingested 30 g specifically produced, intrinsically L-[1-13C]-phenylalanine and L-[1-13C]-leucine labeled lesser mealworm– or milk-derived protein after a unilateral bout of resistance-type exercise. Primed continuous L-[ring-2H5]-phenylalanine, L-[ring-3,5-2H2]-tyrosine, and L-[1-13C]-leucine infusions were applied, with frequent collection of blood and muscle tissue samples. Results A total of 73% ± 7% and 77% ± 7% of the lesser mealworm and milk protein–derived phenylalanine was released into the circulation during the 5 h postprandial period, respectively, with no significant differences between groups (P < 0.05). Muscle protein synthesis rates increased after both lesser mealworm and milk protein concentrate ingestion from 0.025 ± 0.008%/h to 0.045 ± 0.017%/h and 0.028 ± 0.010%/h to 0.056 ± 0.012%/h at rest and from 0.025 ± 0.012%/h to 0.059 ± 0.015%/h and 0.026 ± 0.009%/h to 0.073 ± 0.020%/h after exercise, respectively (all P < 0.05), with no differences between groups (both P > 0.05). Incorporation of mealworm and milk protein-derived L-[1-13C]-phenylalanine into de novo muscle protein was greater after exercise than at rest (P < 0.05), with no differences between groups (P > 0.05). Conclusions Ingestion of a meal-like amount of lesser mealworm–derived protein is followed by rapid protein digestion and amino acid absorption and increases muscle protein synthesis rates both at rest and during recovery from exercise. The postprandial protein handling of lesser mealworm does not differ from ingesting an equivalent amount of milk protein concentrate in vivo in humans. This trial was registered at www.trialregister.nl as NL6897.

Glucosinolates are token stimuli in host selection of many crucifer specialist insects, but the underlying molecular basis for host selection in these insects remains enigmatic. Using a combination of behavioral, electrophysiological, and molecular methods, we investigate glucosinolate receptors in the cabbage butterfly Pieris rapae. Sinigrin, as a potent feeding stimulant, elicited activity in larval maxillary lateral sensilla styloconica, as well as in adult medial tarsal sensilla. Two P. rapae gustatory receptor genes PrapGr28 and PrapGr15 were identified with high expression in female tarsi, and the subsequent functional analyses showed that Xenopus oocytes only expressing PrapGr28 had specific responses to sinigrin; when ectopically expressed in Drosophila sugar sensing neurons, PrapGr28 conferred sinigrin sensitivity to these neurons. RNA interference experiments further showed that knockdown of PrapGr28 reduced the sensitivity of adult medial tarsal sensilla to sinigrin. Taken together, we conclude that PrapGr28 is a gustatory receptor tuned to sinigrin in P. rapae, which paves the way for revealing the molecular basis of the relationships between crucifer plants and their specialist insects.

We investigate how the black soldier fly Hermetia illucens L. (Diptera: Stratiomyidae) responds to dietary protein (P) and carbohydrate (C) contents and the P:C ratio in terms of both immature and adult life-history traits, as well as effects on larval body composition. Nine chicken-feed based diets varying in their P:C ratio are formulated. We test three protein concentrations (10%, 17% and 24%) and three carbohydrate concentrations (35%, 45% and 55%) and their combinations. All nine diets support the complete development and reproduction of this species. Survival is high on all diets. Development time, larval yield, larval crude fat and egg yield are more influenced by P and C contents than by the P:C ratio. Low contents result in a shorter development time. Larval yield is higher on diets with higher C-contents. Pupal development is faster on a low dietary P-content for all three C-contents. Egg yield only increases when P-content increases, although it also varies with the P:C ratio. Larval crude protein content is similar on all nine diets but increases when C-content is low (10%) in P10 and P17. Larval crude fat content is high at P24-diets irrespective of C-content. We conclude that a high macronutrient content combined with a low P:C ratio positively affects H. illucens performance. The diet P17:C55 supports the highest larval and adult performance and results in a high larval body protein content and an intermediate crude fat content.

Global population growth, an increasing demand for animal products and scarcity of conventional feed ingredients drive the search for alternative protein sources for animal feed. Extensive research indicates that insects provide good opportunities as a sustainable, high quality and low-cost component of animal feed. Here, we discuss how insect farming can promote inclusive business for smallholder farmers in the agribusiness value chain. Inclusive business models involving insects as ingredients in feed may contribute to solving socio-economic and environmental problems in developing countries, aligning with the United Nations’ Sustainable Development Goals. With low initial capital investments, smallholder insect farmers have good opportunities to increase productivity, improve their livelihood and contribute to food security and a circular economy.

The acceptance of eco-friendly black soldier fly larvae meal (BSFLM) as sustainable alternative protein ingredient in poultry feeds continues to gain momentum worldwide. This study evaluates the impact of BSFLM in layer chick and grower diets on the growth, carcass quality and economic returns. Mean weekly weight gain and total live weight per chick and grower varied significantly. The highest final weight gain was achieved when birds were provided diet with 25.6% BSFLM. Average daily feed intake (ADFI), average daily weight gain (ADG) and overall weight gain of the chick varied significantly, except for the feed conversion ratio (FCR). For grower birds, ADFI, ADG, FCR and overall weight gain did not vary significantly across the various feeding regimes. The weight of the wings and drumsticks had a quadratic response with a maximum weight obtained at 33% inclusion of BSFLM. The weight of the internal organs were not significantly affected by dietary types. Positive cost–benefit ratio and return on investment was recorded for diet types with higher BSFLM inclusion levels (&gt, 75%). Diets with 25% and 100% BSFLM inclusion were the most suitable and cost-effective, respectively. Thus, BSFLM represents a promising alternative source of protein that could be sustainably used in the poultry industries.

Plant–soil feedback (PSF) may influence plant–insect interactions. Although plant defense differs between shoot and root tissues, few studies have examined root-feeding insect herbivores in a PSF context. We examined here how plant growth and resistance against root-feeding Delia radicum larvae was influenced by PSF. We conditioned soil with cabbage plants that were infested with herbivores that affect D. radicum through plant-mediated effects: leaf-feeding Plutella xylostella caterpillars and Brevicoryne brassicae aphids, root-feeding D. radicum larvae, and/or added rhizobacterium Pseudomonas simiae WCS417r. We analyzed the rhizosphere microbial community, and in a second set of conspecific plants exposed to conditioned soil, we assessed growth, expression of defense-related genes, and D. radicum performance. The rhizosphere microbiome differed mainly between shoot and root herbivory treatments. Addition of Pseudomonas simiae did not influence rhizosphere microbiome composition. Plant shoot biomass, gene expression, and plant resistance against D. radicum larvae was affected by PSF in a treatment-specific manner. Soil conditioning overall reduced plant shoot biomass, Pseudomonas simiae-amended soil causing the largest growth reduction. In conclusion, shoot and root insect herbivores alter the rhizosphere microbiome differently, with consequences for growth and resistance of plants subsequently exposed to conditioned soil.

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.

Adults of many mosquito species feed on plants to obtain metabolic energy and to enhance reproduction. Mosquitoes primarily rely on olfaction to locate plants and are known to respond to a range of plant volatiles. We studied the olfactory response of the yellow fever mosquito Aedes aegypti to methyl jasmonate (MeJA) and cis-jasmone (CiJA), volatile compounds originating from the octadecanoid signaling pathway that plays a key role in plant defense against herbivores. Specifically, we investigated how Ae. aegypti of different ages responded to elevated levels of CiJA in two attractive odor contexts, either derived from Lima bean plants or from human skin. Aedes aegypti females landed significantly less often on a surface with CiJA and MeJA compared to the solvent control, CiJA exerting a stronger reduction in landing than MeJA. Odor context (plant or human) had no significant main effect on the olfactory responses of Ae. aegypti females to CiJA. Mosquito age significantly affected the olfactory response, older females (7–9 d) responding more strongly to elevated levels of CiJA than young females (1–3 d) in either odor context. Our results show that avoidance of CiJA by Ae. aegypti is independent of odor background, suggesting that jasmonates are inherently aversive cues to these mosquitoes. We propose that avoidance of plants with elevated levels of jasmonates is adaptive to mosquitoes to reduce the risk of encountering predators that is higher on these plants, i.e. by avoiding ‘enemy-dense-space’.

Background Semi-field experiments with human landing catch (HLC) measure as the outcome are an important step in the development of novel vector control interventions against outdoor transmission of malaria since they provide good estimates of personal protection. However, it is often infeasible to determine whether the reduction in HLC counts is due to mosquito mortality or repellency, especially considering that spatial repellents based on volatile pyrethroids might induce both. Due to the vastly different impact of repellency and mortality on transmission, the community-level impact of spatial repellents can not be estimated from such semi-field experiments. Methods We present a new stochastic model that is able to estimate for any product inhibiting outdoor biting, its repelling effect versus its killing and disarming (preventing host-seeking until the next night) effects, based only on time-stratified HLC data from controlled semi-field experiments. For parameter inference, a Bayesian hierarchical model is used to account for nightly variation of semi-field experimental conditions. We estimate the impact of the products on the vectorial capacity of the given Anopheles species using an existing mathematical model. With this methodology, we analysed data from recent semi-field studies in Kenya and Tanzania on the impact of transfluthrin-treated eave ribbons, the odour-baited Suna trap and their combination (push–pull system) on HLC of Anopheles arabiensis in the peridomestic area. Results Complementing previous analyses of personal protection, we found that the transfluthrin-treated eave ribbons act mainly by killing or disarming mosquitoes. Depending on the actual ratio of disarming versus killing, the vectorial capacity of An. arabiensis is reduced by 41 to 96% at 70% coverage with the transfluthrin-treated eave ribbons and by 38 to 82% at the same coverage with the push–pull system, under the assumption of a similar impact on biting indoors compared to outdoors. Conclusions The results of this analysis of semi-field data suggest that transfluthrin-treated eave ribbons are a promising tool against malaria transmission by An. arabiensis in the peridomestic area, since they provide both personal and community protection. Our modelling framework can estimate the community-level impact of any tool intervening during the mosquito host-seeking state using data from only semi-field experiments with time-stratified HLC. Graphical Abstract

Plant growth-promoting rhizobacteria (PGPR) can enhance plant growth and defence. Via plant-mediated effects, PGPR have been reported to impact the performance of generalist leaf-chewing insects either negatively or positively. However, only a few insect species, mainly feeding on aboveground tissues, have thus far been investigated. Here, we investigated how addition of rhizobacteria to the soil in which cabbage plants are growing affects the performance of three chewing insect herbivores, two leaf chewers and one root feeder. In a greenhouse experiment, we grew white cabbage (Brassica oleracea) plants in soil supplemented with and without the rhizobacterium Pseudomonas simiae WCS417r. We investigated the consequences for three important insect pests of Brassica species, larvae of the cabbage moth Mamestra brassicae, the diamondback moth Plutella xylostella, or the cabbage root fly Delia radicum after 5 weeks of plant growth. We recorded aboveground plant biomass, insect biomass, plant defence marker gene expression levels and plant defence-related hormone concentrations. Rhizobacterial inoculation increased aboveground plant biomass in non-infested plants but not in infested plants. Rhizobacterial inoculation affected insect performance differently: on PGPR-inoculated plants biomass of Plutella xylostella was lower, while biomass of Delia radicum was higher than on control plants. However, no effect was found on Mamestra brassicae biomass. Rhizobacterial inoculation increased the expression of the defence marker gene LOX2 in P. xylostella-infested plants and MYC2 in M. brassicae-infested plants. Transcription levels of the plant defence marker gene PAL1 showed upregulation between inoculated and non-inoculated insect-free plants. Levels of the phytohormones jasmonic acid, salicylic acid and abscisic acid were similar in inoculated and non-inoculated plants. We conclude that rhizobacterial inoculation has potential to be applied in the protection of cabbage crops against the diamondback moth whereas this does not apply to reducing damage caused by the cabbage moth or cabbage root fly.

There is an increased interest for using insects, such as the black soldier fly, to treat surplus manure and upcycle nutrients into the food system. Understanding the influence that BSFL have on nutrient flows and nutrient losses during manure bioconversion is key for sustainability assessments. Here we quantified and compared nutrient balances, nutrient levels in residual materials and emissions of greenhouse gases and ammonia between manure incubated with black soldier fly larvae (BSFL) and manure without BSFL, during a 9-day experimental period. We obtained high analytical recoveries, ranging between 95 and 103%. We found that of the pig manure supplied, 12.5% of dry matter (DM), 13% of carbon, 25% of nitrogen, 14% of energy, 8.5% of phosphorus and 9% of potassium was stored in BSFL body mass. When BSFL were present, more carbon dioxide (247 vs 148 g/kg of DM manure) and ammonia-nitrogen (7 vs 4.5 g/kg of DM manure) emitted than when larvae were absent. Methane, which was the main contributor to greenhouse gas emissions, was produced at the same levels (1.3 vs 1.1 g/kg of DM manure) in both treatments, indicating the main role that manure microbial methane emissions play. Nitrous oxide was negligible in both treatments. The uptake of nutrients by the larvae and the higher carbon dioxide and ammonia emissions modified the nutrient composition of the residual material substantially relative to the fresh manure. Our study provides a reliable basis to quantify the environmental impact of using BSFL in future life cycle assessments.

Plant‐mediated interactions are an important force in insect ecology. Through such interactions, herbivores feeding on leaves can affect root feeders. However, the mechanisms regulating the effects of above‐ground herbivory on below‐ground herbivores are poorly understood. Here, we investigated the performance of cabbage root fly larvae (Delia radicum) on cabbage plants (Brassica oleracea) previously exposed to above ground herbivores belonging to two feeding guilds: leaf chewing diamondback moth caterpillars (Plutella xylostella) or phloem‐feeding cabbage aphids (Brevicoryne brassicae). Our study focusses on root‐herbivore performance and defence signalling in primary roots by quantifying phytohormones and gene expression. We show that leaf herbivory by caterpillars, but not by aphids, strongly attenuates root herbivore performance. Above‐ground herbivory causes changes in primary roots in terms of gene transcripts and metabolites involved in plant defence. Feeding by below‐ground herbivores strongly induces the jasmonate pathway in primary roots. Caterpillars feeding on leaves cause a slight induction of the primary root jasmonate pathway and interact with plant defence signalling in response to root herbivores. In conclusion, feeding by a leaf chewer and a phloem feeder differentially affects root‐herbivore performance, root‐herbivore‐induced phytohormonal signalling, and secondary metabolites., Caterpillars (Plutella xylostella) and aphids (Brevicoryne brassicae) feeding on leaves of Brassica oleracea differentially affect gene expression, phytohormone concentrations and defence against cabbage root fly larvae (Delia radicum) in the primary root.

The oviposition behavior of mosquitoes is mediated by chemical cues. In the malaria mosquito Anopheles gambiae, conspecific larvae produce infochemicals that affect this behavior. Emanations from first instar larvae proved strongly attractive to gravid females, while those from fourth instars caused oviposition deterrence, suggesting that larval developmental stage affected the oviposition choice of the female mosquito. We examined the nature of these chemicals by headspace collection of emanations of water in which larvae of different stages were developing. Four chemicals with putative effects on oviposition behavior were identified: dimethyldisulfide (DMDS) and dimethyltrisulfide (DMTS) were identified in emanations from water containing fourth instars; nonane and 2,4-pentanedione (2,4-PD) were identified in emanations from water containing both first and fourth instars. Dual-choice oviposition studies with these compounds were done in the laboratory and in semi-field experiments in Tanzania. In the laboratory, DMDS and DMTS were associated with oviposition-deterrent effects, while results with nonane and 2,4-PD were inconclusive. In further studies DMDS and DMTS evoked egg retention, while with nonane and 2,4-PD 88% and 100% of female mosquitoes, respectively, laid eggs. In dual-choice semi-field trials DMDS and DMTS caused oviposition deterrence, while nonane and 2,4-PD evoked attraction, inducing females to lay more eggs in bowls containing these compounds compared to the controls. We conclude that oviposition of An. gambiae is mediated by these four infochemicals associated with conspecific larvae, eliciting either attraction or deterrence. High levels of egg retention occurred when females were exposed to chemicals associated with fourth instar larvae. Electronic supplementary material The online version of this article (10.1007/s10886-020-01175-5) contains supplementary material, which is available to authorized users.

The utilization of the black soldier fly (BSF) Hermetia illucens L. for recycling organic waste into high-quality protein and fat biomass for animal feeds has gained momentum worldwide. However, information on the genetic diversity and environmental implications on safety of the larvae is limited. This study delineates genetic variability and unravels gut microbiome complex of wild-collected and domesticated BSF populations from six continents using mitochondrial COI gene and 16S metagenomics. All sequences generated from the study linked to H. illucens accessions KM967419.1, FJ794355.1, FJ794361.1, FJ794367.1, KC192965.1, and KY817115.1 from GenBank. Phylogenetic analyses of the sequences generated from the study and rooted by GenBank accessions of Hermetia albitarsis Fabricius and Hermetia sexmaculata Macquart separated all samples into three branches, with H. illucens and H. sexmaculata being closely related. Genetic distances between H. illucens samples from the study and GenBank accessions of H. illucens ranged between 0.0091 and 0.0407 while H. sexmaculata and H. albitarsis samples clearly separated from all H. illucens by distances of 0.1745 and 0.1903, respectively. Genetic distance matrix was used to generate a principal coordinate plot that further confirmed the phylogenetic clustering. Haplotype network map demonstrated that Australia, United States 1 (Rhode Island), United States 2 (Colorado), Kenya, and China shared a haplotype, while Uganda shared a haplotype with GenBank accession KC192965 BSF from United States. All other samples analyzed had individual haplotypes. Out of 481,695 reads analyzed from 16S metagenomics, four bacterial families (Enterobactereaceae, Dysgonomonadaceae, Wohlfahrtiimonadaceae, and Enterococcaceae) were most abundant in the BSF samples. Alpha-diversity, as assessed by Shannon index, showed that the Kenyan and Thailand populations had the highest and lowest microbe diversity, respectively; while microbial diversity assessed through Bray Curtis distance showed United States 3 (Maysville) and Netherlands populations to be the most dissimilar. Our findings on genetic diversity revealed slight phylogeographic variation between BSF populations across the globe. The 16S data depicted larval gut bacterial families with economically important genera that might pose health risks to both animals and humans. This study recommends pre-treatment of feedstocks and postharvest measures of the harvested BSF larvae to minimize risk of pathogen contamination along the insect-based feed value chain.

1. Pollinating insects exploit visual and olfactory cues associated with flower traits indicative of flower location and reward quality. Pollination can induce changes in these flower-associated cues, thereby influencing the behaviour of flower visitors. 2. This study investigated the main cues exploited by the syrphid fly Episyrphus balteatus and the butterfly Pieris brassicae when visiting flowers of Brassica nigra and Raphanus sativus plants. Whether pollen is used as a cue and whether pollination-induced changes affect flower volatile emission and the behavioural responses of the two pollinator species were also studied. 3. Pollinator preference was investigated by offering visual and olfactory cues individually as well as simultaneously in two-choice bioassays. Plant treatments included emasculation, hand-pollination and untreated control plants. The composition of flower volatiles from pollinated and unpollinated control plants was analysed by gas chromatography-mass spectrometry. 4. Both pollinators exhibited a strong bias for visual cues over olfactory cues. Neither pollinator used pollen as a cue. However, E. balteatus discriminated between newly opened and long-open flowers at short distance only when pollen was available. Flower visits by pollinators were influenced by pollination-induced changes in B. nigra but not R. sativus flowers. Pieris brassicae only responded to pollination-induced changes when visual and olfactory cues were offered simultaneously. The blend of volatiles emitted by B. nigra, but not R. sativus inflorescences was affected by pollination. 5. Collectively, the findings of this study show that different pollinators exploit different visual and olfactory traits when searching for flowers of two brassicaceous plant species.

The attention for black soldier fly larvae (BSFL) as an alternative ingredient for food and feed products is on the rise. While many studies have reported the efficiency of BSFL to bio‐convert a wide range of organic waste streams into larval biomass, so far, it is unknown whether BSFL prefer certain waste streams over others when they have the possibility to choose. Here, we performed a choice‐test experiment to explore the preference of BSFL when exposed to pig manure and a mass‐rearing diet consisting of plant by‐products currently used for industrial BSFL production. We found that after 1 hr of exposure to both feeds, BSFL strongly preferred pig manure over the mass‐rearing diet. The preference for manure became stronger as larval age increased. Our results provide the first evidence that BSFL express a distinct diet preference. Understanding the reasons for the strong preference for manure is relevant for a diverse array of practical applications and to inform the discussion on insect welfare.

International audience; Edible insects are advocated as sustainable and healthy food and feed. However, commercially produced insects are often low in n-3 fatty acids and have suboptimal n-6/n-3 ratios. A certain amount and proportion of these FAs is required to optimize human health. Flaxseed oil consists primarily (57%) out of alpha-linolenic acid. An experiment was conducted to quantify the effect of flaxseed oil provision on fatty acid composition and to determine the quantity needed to attain a beneficial n-6/n-3 ratio. Three species were used in the experiment: house crickets (Acheta domesticus [L.]), lesser mealworms (Alphitobius diaperinus [Pfanzer]) and black soldier flies (Hermetia illucens [L.]). These were provided with either a control diet or a diet enriched with 1%, 2%, or 4% flaxseed oil during their larval/nymphal stage. Fatty acid profiles of diets and insects were determined via GC-MS. The three species had distinct fatty acid profiles on all four diets, but responded similarly to flaxseed oil addition. For each percent added to the diet, the alpha-linolenic acid content of the insects increased by 2.3%-2.7%. Four percent addition increased the n-3 fatty acid content 10-20 fold in the three species and thereby strongly decreased n-6/n-3 ratios from 18-36 to 0.8-2.4. A ratio below 5 is considered optimal for human health and was achieved by 2% flaxseed oil inclusion for house crickets and lesser mealworms, and at 1% inclusion for black soldier flies. Adding a source of n-3 fatty acids to insect diets can thus improve the nutritional quality of insects.

Edible insects are increasingly being considered as sustainable alternatives to fish and soybean meals in animal feed because of their high nutritional quality and environmental benefits. However, successful introduction of a new product to the market depends on the target user's acceptance. Thus, evaluating the potential demand of insect-based feeds would provide relevant information for policy development. The present study assessed farmers' knowledge on edible insects as feed, their acceptance of integrating insect meals in animal feeds and willingness to pay (WTP) for insect-based feed (IBF) using a contingent valuation method. A household survey was conducted among 957 randomly selected farmers including: 409 poultry, 241 fish and 307 pig farmers in four counties in Kenya. Results of the study reveal that over 70 and 80% of poultry and fish farmers, respectively, are aware that insects can be used as a feed ingredient. In addition, over 60 and 75% of poultry and fish farmers, respectively, consider insects as a good component of feed. Poultry, pig and fish farmers interviewed accepted and showed willingness to pay for IBF. Regression analysis indicated that age, gender, education, marital status, distance to feed trader, awareness of insects as feed, attitude towards insects, acceptance of insect species, availability of agricultural inputs, use of commercial feeds, availability of training and market information had a significant influence on the WTP for IBF. Therefore, increased extension services to educate famers on the nutritional benefits of insect meals in animal feeds and existing market opportunities are expected to improve farmers' attitude towards utilization and consequently enhance WTP for IBF, which in return would significantly reduce the existing pressure on conventional fishmeal feed resources. Our findings provide the first insights into the market opportunities of including insect meals in the animal feed value chain in Kenya.

In addition to controlling pest organisms, the systemic neurotoxic pesticide fipronil can also have adverse effects on beneficial insects and other non-target organisms. Here, we report on the sublethal effects of fipronil on the farmland butterfly Pieris brassicae . Caterpillars were reared on plants that had been grown from seeds coated with fipronil or on leaf discs topically treated with a range of fipronil dosages (1–32 µg kg −1 on dry mass basis). Females that had developed on fipronil plants laid ca half the number of eggs than females that had developed on control plants. In the bioassay with leaf discs, longevity and lifetime egg production declined with increasing fipronil dosage. Remarkably, exposure to fipronil during larval development primarily affected the adult stage. Chemical analyses of leaf tissues collected from seed-treated plants revealed concentrations of fipronil and its degradation products close to the analytical limit of detection (less than or equal to 1 µg kg −1 ). The effective dosage was fivefold higher in the leaf-disc than in the whole-plant experiment. In the whole plant, degradation of fipronil to products that are more toxic than fipronil may explain this discrepancy. Neurotoxicity of insecticides at the level of detection decreases the probability of pinpointing insecticides as the causal agent of harmful effects on non-target organisms.

Black soldier fly larvae (BSFL) are acknowledged for their potential to upcycle waste biomass into animal feed, human food or biofuels. To ensure sustainable BSFL rearing, insight into nutrient bioconversion efficiencies and nutrient losses via gaseous emissions is key. This study used a mass balance approach to quantify nutrient bioconversion efficiencies (i.e., carbon, energy, nitrogen, phosphorus and potassium) and gaseous emissions (i.e., greenhouse gasses and ammonia) of BSFL reared on a substrate used in industrial production. On this substrate, bioconversion efficiencies ranged from 14% (potassium) to 38% (nitrogen). The proportion of dietary inputs found in the residues ranged from 55% (energy) to 86% (potassium), while the proportion of dietary inputs lost via gaseous emissions ranged from 1% (nitrogen) to 24% (carbon). Direct emissions of methane and nitrous oxide during rearing were 16.8 ± 8.6 g CO2-equivalents per kg of dry BSFL biomass. Even though ammonia emissions were minimal, these could have been avoided if larvae would have been harvested before the CO2 peak was reached. Our results provide the first complete mass balance and comprehensive quantification of BSF larval metabolism and GHG emissions, required to assess and improve the environmental sustainability of BSFL production systems.