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Tettamanti, G.

Publications and source records attributed to Tettamanti, G..

2 recordsLinked to original sources

The gut bacterial community of black soldier fly larvae is a reservoir of antibiotic resistance and virulence genes

1.Antimicrobial resistance (AMR) is a serious threat to global health. Agricultural practices that have contributed greatly to AMR spread urgently require innovation to address this issue, and more broadly challenges of sustainability and environmental concern. The larvae of black soldier fly (BSFL), Hermetia illucens, are considered a promising resource for advancing sustainable and circular agri-food systems given their ability to bioconvert organic waste streams into protein-and lipid-rich biomass suitable for feed applications and the use of the rearing residues (i.e., frass) as organic fertilisers. However, despite their emerging industrial applications, the risks of antibiotic resistance spread through their use remain underexplored. To elucidate this aspect, the profiles of antibiotic resistance genes (ARGs) and virulence factors (VFs), and their occurrence on plasmids were predicted from the midgut bacterial community of BSFL. Shotgun metagenomics revealed candidate resistance genes for 26 classes of antibiotics, and virulence via 9 mechanisms (with mobility and biofilm formation as major ones), with taxa belonging to the Pseudomonadota phylum as the dominant contributors. Highly relevant to public health was the identification of genes encoding resistance to carbapenem class antibiotics in bacterial genomes and mobile plasmids. Reconstruction of metagenomes enabled more precise taxonomic resolution and revealed taxa harbouring multiple resistance and virulence genes, including a Pseudomonas species with 42 VFs and 7 ARGs. Notably, for the first time antibiotic resistant bacterial species were isolated from the gut microbiota of BSFL, validating and complementing the results obtained in silico. Together, this work represents a comprehensive profile of the BSFL midgut bacterial resistome, while also providing relevant context on virulence and mobility. Importantly, it emphasises the urgent need to adopt strategies to mitigate potential risks arising from the development of emerging technologies related to the use of insect-mediated bioconversion and derived products.

genomics↗

Drosophila melanogaster as a platform for the functional expression of engineered PET-degrading enzymes

Insects offer promising opportunities for organic waste bioconversion; however, they cannot efficiently degrade synthetic polymers such as polyethylene terephthalate (PET). Here, we generated transgenic Drosophila melanogaster lines to express in vitro-evolved variants of two PET-degrading enzymes with distinct biochemical properties: an engineered Ideonella sakaiensis PETase variant (TS-{Delta}IsPET) and a leaf-branch compost cutinase variant (TA-{Delta}LCC). Both enzymes, fused to a Drosophila gut-derived secretory signal, were produced and secreted by both Drosophila cultured S2R+ cells and transgenic larvae. Both enzymes were glycosylated upon secretion, a post-translational modification that did not abolish their catalytic activity. Notably, TA-{Delta}LCC displayed [~]6-fold higher esterase activity than TS-{Delta}IsPET in larval extracts and TA-{Delta}LCC-containing extracts depolymerised PET nanoparticles in vitro under enzyme-favourable conditions. Transgenic flies showed normal development, fertility and survival. Morphological and biochemical analysis confirmed that TA-{Delta}LCC expression did not alter midgut structure and function. Together, these results establish Drosophila melanogaster as a model for functional expression and comparative evaluation of engineered PET-degrading enzymes and identify TA-{Delta}LCC as a promising candidate for exploitation in insect species relevant to plastic contaminated waste bioconversion. HighlightsO_LITransgenic D. melanogaster enables in vivo study of engineered PET enzymes C_LIO_LIEngineered TS-{Delta}IsPET and TA-{Delta}LCC are functional in larval extracts C_LIO_LITA-{Delta}LCC was selected for PET nanoparticle assays due to higher pNPA activity C_LIO_LID. melanogaster model enables comparative evaluation of PET-degrading enzymes C_LI

biochemistry↗