Nitrosative stress defences of the enterohepatic pathogenic bacterium Helicobacter pullorum

Helicobacter pullorum is an avian bacterium that causes gastroenteritis, intestinal bowel and hepatobiliary diseases in humans. Although H. pullorum has been shown to activate the mammalian innate immunity with release of nitric oxide (NO), the proteins that afford protection against NO and reactive nitrogen species (RNS) remain unknown. Here several protein candidates of H. pullorum, namely a truncated (TrHb) and a single domain haemoglobin (SdHb), and three peroxiredoxin-like proteins (Prx1, Prx2 and Prx3) were investigated. We report that the two haemoglobin genes are induced by RNS, and that SdHb confers resistance to nitrosative stress both in vitro and in macrophages. For peroxiredoxins, the prx2 and prx3 expression is enhanced by peroxynitrite and hydrogen peroxide, respectively. Mutation of prx1 does not alter the resistance to these stresses, while the single ∆prx2 and double ∆prx1∆prx2 mutants have decreased viability. To corroborate the physiological data, the biochemical analysis of the five recombinant enzymes was done, namely by stopped-flow spectrophotometry. It is shown that H. pullorum SdHb reacts with NO much more quickly than TrHb, and that the three Prxs react promptly with peroxynitrite, Prx3 displaying the highest reactivity. Altogether, the results unveil SdHb and Prx3 as major protective systems of H. pullorum against nitrosative stress

Lipids from Insects in Cosmetics and for Personal Care Products

Insects, the most varied group of known organisms on Earth, are arousing great interest also for the possibility to use them as a feed and food source. The mass rearing of some species, defined as “bioconverters”, is spreading worldwide, thanks to their sustainability. At the end of the bioconversion process, breeders obtain eco-friendly biomolecules of high biological and economic value, including proteins and lipids, from larvae of bioconverter insects, in particular Hermetia illucens. Besides the most classical use of insect lipids as food additives, they are also used in the formulation of several products for personal care. The composition of insect lipids depends on the substrate on which the insects are reared but also on the insect species, so the cosmetic producers should consider these features to choose their insect starting point. The most abundant fatty acids detected in H. illucens are lauric, myristic, palmitic, and oleic acids, regardless of feed substrate; its fatty acids composition is favorable for soap composition, while their derivatives are used for detergent and shampoo. Here, we offer an overview of insect lipids, their extraction methods, and their application in cosmetics and personal care products

Current state of chitin purification and chitosan production from insects

Chitin, and especially its deacetylated variant chitosan, has many applications, e.g. as carrier material for pharmaceutical drugs or as a flocculant in wastewater treatment. Despite its versatility and accessibility, chitin, the second most abundant polysaccharide on Earth, has so far been commercially extracted only from crustaceans and to a minor extent from fungi. Insects are a viable alternative source of chitin, but they have not been exploited in the past due to limited availability. Today however, for the sustainable production of animal feed, insect farming is being developed substantially. The availability of large quantities of insect biomass and chitin-rich side products such as exuviae and exoskeletons has been increasing. This review provides an overview of recently published studies of chitin extraction from insects, its subsequent conversion into chitosan and the primary analytical methods used to characterize insect-based chitin and chitosan. We have discovered a large number of research articles published over the past 20 years, confirming the increased attention being received by chitin and chitosan production from insects. Despite numerous publications, we identified several knowledge gaps, such as a lack of data concerning chitin purification degree and chitosan yield. Furthermore, analytical methods used to obtain physicochemical characteristics, structural information and chemical composition meet basic qualitative requirements but do not satisfy the need for a more quantitative evaluation. Despite the current shortcomings that need to be overcome, this review presents encouraging data on the use of insects as an alternative source of chitin and chitosan in the future. © 2020 The Authors. Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI)

Amyloid/Melanin distinctive mark in invertebrate immunity

Protostomes and Deuterostomes show the same nexus between melanin production, and amyloid fibril production, i.e., the presence of melanin is indissolubly linked to amyloid scaffold that, in turn, is conditioned by the redox status/cytoplasmic pH modification, pro-protein cleavage presence, adrenocorticotropin hormone (ACTH), melanocyte-stimulating hormone (\u3b1-MSH), and neutral endopeptidase (NEP) overexpressions. These events represent the crucial component of immune response in invertebrates, while in vertebrates these series of occurrences could be interpreted as a modest and very restricted innate immune response. On the whole, it emerges that the mechanisms involving amyloid fibrils/pigment synthesis in phylogenetically distant metazoan (viz, cnidaria, molluscs, annelids, insects, ascidians and vertebrates) are evolutionary conserved. Furthermore, our data show the relationship between immune and neuroendocrine systems in amyloid/melanin synthesis. Indeed the process is closely associated to ACTH-\u3b1-MSH production, and their role in stress responses leading to pigment production reflects and confirms again their ancient phylogeny

Structural and functional characterization of a novel recombinant antimicrobial peptide from hermetia illucens

Antibiotics are commonly used to treat pathogenic bacteria, but their prolonged use con-tributes to the development and spread of drug-resistant microorganisms raising the challenge to find new alternative drugs. Antimicrobial peptides (AMPs) are small/medium molecules ranging 10–100 residues synthesized by all living organisms and playing important roles in the defense sys-tems. These features, together with the inability of microorganisms to develop resistance against the majority of AMPs, suggest that these molecules might represent effective alternatives to clas-sical antibiotics. Because of their high biodiversity, with over one million described species, and their ability to live in hostile environments, insects represent the largest source of these molecules. However, production of insect AMPs in native forms is challenging. In this work we investigate a defensin-like antimicrobial peptide identified in the Hermetia illucens insect through a combination of transcriptomics and bioinformatics approaches. The C-15867 AMP was produced by recombi-nant DNA technology as a glutathione S-transferase (GST) fusion peptide and purified by affinity chromatography. The free peptide was then obtained by thrombin proteolysis and structurally characterized by mass spectrometry and circular dichroism analyses. The antibacterial activity of the C-15867 peptide was evaluated in vivo by determination of the minimum inhibitory concentration (MIC). Finally, crystal violet assays and SEM analyses suggested disruption of the cell membrane architecture and pore formation with leaking of cytosolic material

A mobile black soldier fly farm for on-site disposal of animal dairy manure

Black soldier fly (BSF), Hermetia illucens (L.) (Diptera Stratiomyidae), is a saprophagous insect that is receiving a growing scientific and economic interest since during the larval stage it is extremely voracious and able to consume a wide range of organic materials. This ethological characteristic is particularly suitable for waste management at industrial scale. The extraordinary ability to accumulate high levels of proteins and lipids, allows the use of resulting larvae as animal feed or biodiesel production; the residue of the bioconversion process, that consists of larval frass and not converted organic matter is assimilable to organic fertilizer. The aim of this study was to evaluate the bioconversion process by black soldier fly larvae (BSFL) on fresh and mature dairy manure. A “mobile bioconversion unit” that works as a mobile breeding unit was used, allowing to carry out bioconversion tests directly on fields, in different livestock farms located on the Basilicata territory (Italy). Total larval and frass biomass, bioconversion yield, BSFL development time and substrate reduction were evaluated for each treatment. All the analysed parameters differed from the control (larvae fed in standard diet) but not between the two substrates from the zootechnical chain. Although development time significantly differed across treatments, BSF correctly grow and reduce all substrates confirming larvae can be used to bioconvert animal manure reducing the undesired effects occurring from mismanaged this kind of substrate