Aurora Montali

The saprophagous larvae of Hermetia illucens show promising potential as effective agents for bioconverting organic waste and by-products into valuable biomass. Their capability to efficiently transform organic substrates, varying in nutrient content, origin, texture, and moisture, is strictly correlated with the morphofunctional complexity of their digestive system and the associated microbiota. In addition, post-ingestion mechanisms are set in motion to regulate the midgut activity depending on the feeding substrate, thus ensuring the nutritional requirements of the larvae. This study aims to assess the capability of H. illucens larvae to grow and biotransform the organic fraction of municipal solid waste (OFMSW). In particular, the larvae were reared on substrates mimicking the OFMSW with different nutrient content, and their growth and bioconversion performance, as well as the quality of larval and pupal biomass were evaluated. Our results demonstrate that the nutritional composition of the rearing substrates had minimal impact on the efficiency of the bioconversion process and on the protein, lipid, and chitin content of the insect biomass. This was due to a fine transcriptional regulation of genes encoding enzymes involved in protein, polysaccharide, and lipid digestion in midgut cells. As a consequence, larvae reared on OFMSW with the lowest protein, starch, and lipid content exhibited a significant increase in protease, amylase, and lipase activities. Overall, this study highlights the value of H. illucens larvae in valorising the OFMSW, demonstrating that variations in its composition do not significantly affect the quality of the bioconversion process. These findings hold significant practical implications as current strategies for OFMSW treatment remain unsatisfactory and innovative approaches are needed to enhance bio-waste recycling and reduce the environmental impact of waste management.

BACKGROUND: Despite their known negative effects on ecosystems and human health, synthetic pesticides are still largely used to control crop insect pests. Currently, the biopesticide market for insect biocontrol mainly relies on the entomopathogenic bacterium Bacillus thuringiensis (Bt). New biocontrol tools for crop protection might derive from fungi, in particular from Trichoderma spp., which are known producers of chitinases and other bioactive compounds able to negatively affect insect survival.RESULTS: In this study, we first developed an environmentally sustainable production process for obtaining chitinases from Trichoderma asperellum ICC012. Then, we investigated the biological effects of this chitinase preparation – alone or in combination with a Bt‐based product – when orally administered to two lepidopteran species. Our results demonstrate that T. asperellum efficiently produces a multi‐enzymatic cocktail able to alter the chitin microfibril network of the insect peritrophic matrix, resulting in delayed development and larval death. The co‐administration of T. asperellum chitinases and sublethal concentrations of Bt toxins increased larval mortality. This synergistic effect was likely due to the higher amount of Bt toxins that passed the damaged peritrophic matrix and reached the target receptors on the midgut cells of chitinase‐treated insects.CONCLUSION: Our findings may contribute to the development of an integrated pest management technology based on fungal chitinases that increase the efficacy of Bt‐based products, mitigating the risk of Bt‐resistance development. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Glycopeptide antibiotics (GPAs) are drugs of last resort for treating infections by Gram-positive bacteria. They inhibit bacterial cell wall assembly by binding to the d-Ala-d-Ala terminus of peptidoglycan precursors, leading to cell lysis. Vancomycin and teicoplanin are first generation GPAs, while dalbavancin is one of the few, recently approved, second generation GPAs. In this paper, we developed an in vivo insect model to compare, for the first time, the efficacy of these three GPAs in curing Staphylococcus aureus infection. Differently from previous reports, Bombyx mori larvae were reared at 37 °C, and the course of infection was monitored, following not only larval survival, but also bacterial load in the insect body, hemocyte activity, phenoloxidase activity, and antimicrobial peptide expression. We demonstrated that the injection of S. aureus into the hemolymph of B. mori larvae led to a marked reduction of their survival rate within 24–48 h. GPAs were not toxic to the larvae and cured S. aureus infection. Dalbavancin was more effective than first generation GPAs. Due to its great advantages (i.e., easy and safe handling, low rearing costs, low antibiotic amount needed for the tests, no restrictions imposed by ethical and regulatory issues), this silkworm infection model could be introduced in preclinical phases—prior to the use of mice—accelerating the discovery/development rate of novel GPAs.

Microbial chitinases are gaining interest as promising candidates for controlling plant pests. These enzymes can be used directly as biocontrol agents as well as in combination with chemical pesticides or other biopesticides, reducing their environmental impact and/or enhancing their efficacy. Chitinolytic enzymes can target two different structures in insects: the cuticle and the peritrophic matrix (PM). PM, formed by chitin fibrils connected to glycoproteins and proteoglycans, represents a physical barrier that plays an essential role in midgut physiology and insect digestion, and protects the absorptive midgut epithelium from food abrasion or pathogen infections. In this paper, we investigate how two recently discovered metagenome-sourced chitinases (Chi18H8 and 53D1) affect, in vitro and in vivo, the PM integrity of Bombyx mori, a model system among Lepidoptera. The two chitinases were produced in Escherichia coli or, alternatively, in the unconventional – but more environmentally acceptable – Streptomyces coelicolor. Although both the proteins dramatically altered the structure of B. mori PM in vitro, when administered orally only 53D1 caused adverse and marked effects on larval growth and development, inducing mortality and reducing pupal weight. These in vivo results demonstrate that 53D1 is a promising candidate as insecticide protein.

Pseudomonas aeruginosa is an opportunistic bacterium causing severe infections in hospitalised and immunosuppressed patients, particularly individuals affected by cystic fibrosis. Several clinically isolated P. aeruginosa strains were found to be resistant to three or more antimicrobial classes indicating the importance of identifying new antimicrobials active against this pathogen. Here we characterised the antimicrobial activity and the action mechanisms against P. aeruginosa of two natural isoforms of the antimicrobial peptide Cecropin B, both isolated from the silkworm Bombyx mori. These Cecropin B isoforms differ in a single amino acid substitution within the active portion of the peptide, so that a Glutamic acid of the E53 CecB variant is replaced by a Glutamine in the Q53 CecB isoform. Both peptides showed a high antimicrobial and membranolytic activity against P. aeruginosa, with Q53 CecB displaying greater activity compared to the E53 CecB isoform. Biophysical analyses, live-cell NMR and molecular dynamic simulation studies indicated that both peptides might act as membrane interacting elements, which can disrupt the outer membrane organisation, facilitating their translocation towards the inner membrane of the bacterial cell. Our data also suggest that the amino acid variation of the Q53 CecB isoform represents a critical factor in stabilising the hydrophobic segment that interacts with the bacterial membrane, determining the highest antimicrobial activity of the whole peptide. High stability to pH and temperature variations, the tolerance to high salt concentrations, and the low toxicity against human cells make Q53 CecB a promising candidate in the development of CecB-derived compounds against P. aeruginosa.

Morphogenetic events that occur during development and regeneration are energy demanding processes requiring profound rearrangements in cell architecture, which need to be coordinated in timely fashion with other cellular activities, such as proliferation, migration and differentiation. In the last 15 years, it has become evident that autophagy, an evolutionarily-conserved catabolic process that mediates the lysosomal turnover of organelles and macromolecules, is an essential “tool” to ensure remodelling events that occur at cellular and tissue levels. Indeed, studies in several model organisms have shown that the inactivation of autophagy genes has a significant impact on embryogenesis and tissue regeneration, leading to extensive cell death and persistence of unnecessary cell components. Interestingly, the increased understanding of the mechanisms that confers selectivity to the autophagic process has also contributed to identifying development-specific targets of autophagy across species. Moreover, alternative ways to deliver materials to the lysosome, such as microautophagy, are also emerging as key actors in these contexts, providing a more complete view of how the cell component repertoire is renovated. In this review, we discuss the role of different types of autophagy in development and regeneration of invertebrates and vertebrates, focusing in particular on its contribution in cnidarians, platyhelminthes, nematodes, insects, zebrafish and mammals.

In holometabolous insects, various larval organs are remodeled by autophagy during metamorphosis. Although moths and butterflies are among the first animal models in which this self-eating process was described, only in recent years autophagy has been analyzed in detail in these insects. In particular, the silkworm Bombyx mori, which represents a well-studied model among Lepidoptera, provides a wide repertoire of cellular and molecular tools useful for studying the occurrence of autophagy and for evaluating its role in postembryonic development. Here, we describe some morphological, biochemical, and molecular methods to monitor autophagy in silkworm organs.