Primary study on frass fertilizers from mass-reared insects: Species variation, heat treatment effects, and implications for soil application at laboratory scale.

Insect farming has gained popularity as a resource-efficient and eco-friendly method for managing organic wastes by converting them into high-quality protein, fat, and frass. Insect frass is a powerful organic fertilizer that enriches the soil with essential plant nutrients and enhances plant defense mechanisms through chitin stimulation. Given the importance of frass commercialization for many insect farmers and the use of increasingly diverse organic wastes as insect feedstocks, there is a need for legal guidelines to enable clean production practices. The recent introduction of a legal definition for frass and heat treatment requirements by the EU commission marks a significant step towards standardizing its quality; however, little is known about the processes shaping its nutritional profiles and contributing to its maturation. Our study addresses key knowledge gaps in frass composition and production practices. Here, we analyzed the physicochemical, plant-nutritional, and microbiological properties of black soldier fly, yellow mealworm, and Jamaican field cricket frass from mass-rearing operations and assessed the impact of hygienizing heat treatment on fertilizer properties and frass microbiota. The results showed that frass properties varied significantly across insect species, revealing concentrations of plant-available nutrients as high as 7000 µg NH4+-N, 150 µg NO2-NO3--N, and 20 mg available P per g of total solids. Heat treatment reduced microbial activity, biomass, and viable counts of pathogenic Escherichia coli and Salmonella spp. In terms of frass microbiome composition, alpha diversity showed no significant differences between fresh and heat-treated frass samples; however, significant differences in microbial community composition were observed across the three insect species. Despite heat treatment, soil application of frass reactivated and boosted soil microbial activity, inducing up to a 25-fold increase in microbial respiration, suggesting no long-term detrimental effects on microorganisms. These findings not only enhance our understanding of insect frass as a nutrient-rich organic fertilizer but also have implications for regulatory frameworks, underscoring its promising potential for soil health and nutrient cycling. However, it is important to recognize the primary nature of this research, conducted at laboratory scale and over a short term. Future studies should aim to validate these findings in agricultural settings and explore additional factors influencing frass properties and its (long-term) interaction with soil ecosystems.

Insect frass and exuviae to promote plant growth and health.

Beneficial soil microorganisms can contribute to biocontrol of plant pests and diseases, induce systemic resistance (ISR) against attackers, and enhance crop yield. Using organic soil amendments has been suggested to stimulate the abundance and/or activity of beneficial indigenous microbes in the soil. Residual streams from insect farming (frass and exuviae) contain chitin and other compounds that may stimulate beneficial soil microbes that have ISR and biocontrol activity. Additionally, changes in plant phenotype that are induced by beneficial microorganisms may directly influence plant-pollinator interactions, thus affecting plant reproduction. We explore the potential of insect residual streams derived from the production of insects as food and feed to promote plant growth and health, as well as their potential benefits for sustainable agriculture.

Direct and Indirect Effect via Endophytism of Entomopathogenic Fungi on the Fitness of Myzus persicae and Its Ability to Spread PLRV on Tobacco.

Aphids are major crop pests that transmit more than half of all insect-vectored plant viruses responsible for high yield losses worldwide. Entomopathogenic fungi (EPF) are biological control agents mainly used by foliar application to control herbivores, including sap-sucking pests such as aphids. Their ability to colonize plant tissues and to interact with diverse plant pathogenic microorganisms have been reported. In our study, we evaluated the effectiveness of Beauveria bassiana ((Balsamo-Crivelli) Vuillemin) directly applied by contact or/and indirectly via endophytism in tobacco plants (Nicotiana tabacum L.) against the virus vector Myzus persicae (Sulzer) carrying the Potato leafroll virus (PLRV) or not. We found that both contact treatment and endophytic colonization of leaves significantly increased aphid mortality and decreased the fecundity rate when compared to control plants. In addition, on fungal-colonized leaves, viruliferous aphids were more negatively impacted than virus-free ones and nymph mortality was significantly higher than on fungal-free plants. Furthermore, we assessed PLRV transmission by M. persicae on tobacco plants inoculated with either B. bassiana or Metarhizium acridum ((Driver and Milner) JF Bischoff, Rehner, and Humber) as source or/and recipient plants. Myzus persicae was found to acquire and transmit PLRV regardless of the treatment. Nevertheless, the infection rate of endophytically colonized plants was lower at a seven-day incubation period and had increased to almost 100% after fifteen days. These results suggest that B. bassiana is effective against aphids, both by contact and via endophytism, and both B. bassiana and M. acridum delayed PLRV infection in tobacco.

Does the Infectious Status of Aphids Influence Their Preference Towards Healthy, Virus-Infected and Endophytically Colonized Plants?

Aphids (Hemiptera: Aphididae) cause significant damage and transmit viruses to various crop plants. We aimed to evaluate how the infectious status of aphids influences their interaction with potential hosts. Two aphid (Myzus persicae and Rhopalosiphum padi) and plant (Nicotiana tabacum and Triticum aestivum) species were used. The preferences of aphids towards healthy, virus-infected (Potato Leafroll Virus (PLRV) and Barley Yellow Dwarf virus (BYDV)), and endophytic entomopathogenic fungi (EEPF)-inoculated (Beauveria bassiana and Metarhizium acridum) plants were investigated in dual-choice tests. The headspace volatiles of the different plant modalities were also sampled and analyzed. Viruliferous and non-viruliferous aphids were more attracted to EEPF-inoculated plants compared to uninoculated plants. However, viruliferous aphids were more attracted to EEPF-inoculated plants compared to virus-infected plants, while non-viruliferous insects exhibited no preference. Fungal-inoculated plants released higher amounts of aldehydes (i.e., heptanal, octanal, nonanal and decanal) compared to other plants, which might explain why viruliferous and non-viruliferous aphids were more abundant in EEPF-inoculated plants. Our study provides an interesting research perspective on how EEPF are involved in behavior of virus vector, depending on the infectious status of the latter.

Predatory Earwigs are Attracted by Herbivore-Induced Plant Volatiles Linked with Plant Growth-Promoting Rhizobacteria.

Plant-associated microbes may induce plant defenses against herbivores. Plants, in turn, can attract natural enemies, such as predators, using herbivore-induced plant volatiles. Intricate communication occurs between microorganisms, plants, and insects. Given that many aspects related to mechanisms involved in this symbiotic system remain unknown, we evaluated how beneficial soil-borne microorganisms can affect the interactions between plants, herbivores, and natural enemies. For this study, we established a multitrophic system composed of the predatory earwig Doru luteipes (Dermaptera: Forficulidae), arugula (Eruca sativa, Brassicaceae) as the host plant, Plutella xylostella (Lepidoptera: Plutellidae) larvae as a specialist herbivore, Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae as a generalist herbivore, and Bacillus amyloliquefaciens as the plant growth-promoting rhizobacteria (PGPR), in a series of nocturnal olfactometry experiments. By assessing earwig preference towards herbivore-induced and PGPR-inoculated plants in different combinations, we showed that the interaction between rhizobacteria, plants, and herbivores can affect the predatory earwig's behavior. Furthermore, we observed a synergistic effect in which earwigs were attracted by plants that presented as PGPR inoculated and herbivore damaged, for both specialist and generalist herbivores. Our findings help fill the important knowledge gap regarding multitrophic interactions and should provide useful guidelines for their application to agricultural fields.

High levels of arbuscular mycorrhizal fungus colonization on Medicago truncatula reduces plant suitability as a host for pea aphids (Acyrthosiphon pisum).

This study sheds light on a poorly understood area in insect-plant-microbe interactions, focusing on aphid probing and feeding behavior on plants with varying levels of arbuscular mycorrhizal (AM) fungus root colonization. It investigates a commonly occurring interaction of three species: pea aphid Acyrthosiphon pisum, barrel medic Medicago truncatula, and the AM fungus Rhizophagus irregularis, examining whether aphid-feeding behavior changes when insects feed on plants at different levels of AM fungus colonization (42% and 84% root length colonized). Aphid probing and feeding behavior was monitored throughout 8 h of recording using the electrical penetration graph (EPG) technique, also, foliar nutrient content and plant growth were measured. Summarizing, aphids took longer to reach their 1st sustained phloem ingestion on the 84% AM plants than on the 42% AM plants or on controls. Less aphids showed phloem ingestion on the 84% AM plants relative to the 42% AM plants. Shoots of the 84% AM plants had higher percent carbon (43.7%) relative to controls (40.5%), and the 84% AM plants had reduced percent nitrogen (5.3%) relative to the 42% AM plants (6%). In conclusion, EPG and foliar nutrient data support the hypothesis that modifications in plant anatomy (e.g., thicker leaves), and poor food quality (reduced nitrogen) in the 84% AM plants contribute to reduced aphid success in locating phloem and ultimately to differences in phloem sap ingestion. This work suggests that M. truncatula plants benefit from AM symbiosis not only because of increased nutrient uptake but also because of reduced susceptibility to aphids.