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)

Insect Chitin-Based Nanomaterials for Innovative Cosmetics and Cosmeceuticals. Cosmetics

Chitin and its derivatives are attracting great interest in cosmetic and cosmeceutical fields, thanks to their antioxidant and antimicrobial properties, as well as their biocompatibility and biodegradability. The classical source of chitin, crustacean waste, is no longer sustainable and fungi, a possible alternative, have not been exploited at an industrial scale yet. On the contrary, the breeding of bioconverting insects, especially of the Diptera Hermetia illucens, is becoming increasingly popular worldwide. Therefore, their exoskeletons, consisting of chitin as a major component, represent a waste stream of facilities that could be exploited for many applications. Insect chitin, indeed, suggests its application in the same fields as the crustacean biopolymer, because of its comparable commercial characteristics. This review reports several cosmetic and cosmeceutical applications based on chitin and its derivatives. In this context, chitin nanofibers and nanofibrils, produced from crustacean waste, have proved to be excellent cosmeceutical active compounds and carriers of active ingredients in personal care. Consequently, the insect-based chitin, its derivatives and their complexes with hyaluronic acid and lignin, as well as with other chitin-derived compounds, may be considered a new appropriate potential polymer to be used in cosmetic and cosmeceutical fields. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

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

What antarctic plants can tell us about climate changes: Temperature as a driver for metabolic reprogramming

Global warming is strongly affecting the maritime Antarctica climate and the consequent melting of perennial snow and ice covers resulted in increased colonization by plants. Colobanthus quitensis is a vascular plant highly adapted to the harsh environmental conditions of Antarctic Peninsula and understanding how the plant is responding to global warming is a new challenging target for modern cell physiology. To this aim, we performed differential proteomic analysis on C. quitensis plants grown in natural conditions compared to plants grown for one year inside open top chambers (OTCs) which determine an increase of about 4 °C at midday, mimicking the effect of global warming. A thorough analysis of the up and downregulated proteins highlighted an extensive metabolism reprogramming leading to enhanced photoprotection and oxidative stress control as well as reduced content of cell wall components. Overall, OTCs growth seems to be advantageous for C. quitensis plants which could benefit from a better CO2 diffusion into the mesophyll and a reduced ROS‐mediated photodamage

Identification and Functional Characterization of Toxoneuron nigriceps Ovarian Proteins Involved in the Early Suppression of Host Immune Response

The endophagous parasitoid Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) of the larval stages of the tobacco budworm Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae) injects the egg, the venom, the calyx fluid, which includes a Polydnavirus (T. nigriceps BracoVirus: TnBV) and the Ovarian Proteins (OPs) into the host body during oviposition. The host metabolism and immune system are disrupted prematurely shortly after parasitization by the combined action of the TnBV, venom, and OPs. OPs are involved in the early suppression of host immune response, before TnBV infects and expresses its genes in the host tissues. In this work, we evaluated the effect of HPLC fractions deriving from in toto OPs. Two fractions caused a reduction in hemocyte viability and were subsequently tested to detect changes in hemocyte morphology and functionality. The two fractions provoked severe oxidative stress and actin cytoskeleton disruption, which might explain the high rate of hemocyte mortality, loss of hemocyte functioning, and hence the host\u2019s reduced hemocyte encapsulation ability. Moreover, through a transcriptome and proteomic approach we identify the proteins of the two fractions: eight proteins were identified that might be involved in the observed host hemocyte changes. Our findings will contribute to a better understanding of the secreted ovarian components and their role in parasitoid wasp strategy for evading host immune responses

The Lepidopteran endoribonuclease-U domain protein P102 displays dramatically reduced enzymatic activity and forms functional amyloids

Hemocytes of Heliothis virescens (F.) (Lepidoptera, Noctuidae) larvae produce a protein, P102, with a putative endoribonuclease-U domain. In previous works we have shown that P102 is involved in Lepidopteran immune response by forming amyloid fibrils, which catalyze and localize melanin deposition around non-self intruders during encapsulation, preventing harmful systemic spreading. Here we demonstrate that P102 belongs to a new class of proteins that, at least in Lepidoptera, has a diminished endoribonuclease-U activity probably due to the lack of two out of five catalytically essential residues. We show that the P102 homolog from Trichoplusia ni (Lepidoptera, Noctuidae) displays catalytic site residues identical to P102, a residual endoribonuclease-U activity and the ability to form functional amyloids. On the basis of these results as well as sequence and structural analyses, we hypothesize that all the Lepidoptera endoribonuclease-U orthologs with catalytic site residues identical to P102 form a subfamily with similar function

Nox1/Ref-1-mediated activation of CREB promotes Gremlin1-driven endothelial cell proliferation and migration.

Pulmonary arterial hypertension (PAH) is a complex degenerative disorder marked by aberrant vascular remodeling associated with hyperproliferation and migration of endothelial cells (ECs). Previous reports implicated bone morphogenetic protein antagonist Gremlin 1 in this process; however, little is known of the molecular mechanisms involved. The current study was designed to test whether redox signaling initiated by NADPH oxidase 1 (Nox1) could promote transcription factor CREB activation by redox factor 1 (Ref-1), transactivation of Gremlin1 transcription, EC migration, and proliferation. Human pulmonary arterial EC (HPAECs) exposed in vitro to hypoxia to recapitulate PAH signaling displayed induced Nox1 expression, reactive oxygen species (ROS) production, PKA activity, CREB phosphorylation, and CREB:CRE motif binding. These responses were abrogated by selective Nox1 inhibitor NoxA1ds and/or siRNA Nox1. Nox1-activated CREB migrated to the nucleus and bound to Ref-1 leading to CREB:CRE binding and Gremlin1 transcription. CHiP assay and CREB gene-silencing illustrated that CREB is pivotal for hypoxia-induced Gremlin1, which, in turn, stimulates EC proliferation and migration. In vivo, participation of Nox1, CREB, and Gremlin1, as well as CREB:CRE binding was corroborated in a rat PAH model. Activation of a previously unidentified Nox1-PKA-CREB/Ref-1 signaling pathway in pulmonary endothelial cells leads to Gremlin1 transactivation, proliferation and migration. These findings reveal a new signaling pathway by which Nox1 via induction of CREB and Gremlin1 signaling contributes to vascular remodeling and provide preclinical indication of its significance in PAH

Role of Ovarian Proteins Secreted by Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) in the Early Suppression of Host Immune Response

Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) is an endophagous parasitoid of the larval stages of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae). During oviposition, T. nigriceps injects into the host body, along with the egg, the venom, the calyx fluid, which contains a Polydnavirus (T. nigriceps BracoVirus: TnBV), and the Ovarian Proteins (OPs). Although viral gene expression in the host reaches detectable levels after a few hours, a precocious disruption of the host metabolism and immune system is observed right after parasitization. This alteration appears to be induced by female secretions including TnBV venom and OPs. OPs, originating from the ovarian calyx cells, are involved in the induction of precocious symptoms in the host immune system alteration. It is known that OPs in braconid and ichneumonid wasps can interfere with the cellular immune response before Polydnavirus infects and expresses its genes in the host tissues. Here we show that T. nigriceps OPs induce several alterations on host haemocytes that trigger cell death. The OP injection induces an extensive oxidative stress and a disorganization of actin cytoskeleton and these alterations can explain the high-level of haemocyte mortality, the loss of haemocyte functionality, and so the reduction in encapsulation ability by the host

Rearing substrate impacts growth and macronutrient composition of Hermetia illucens (L.) (Diptera: Stratiomyidae) larvae produced at an industrial scale

Organic waste is a rapidly increasing problem due to the growth of the agricultural production needed to meet global food demands. Development of sustainable waste management solutions is essential. Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) (BSF), larvae are voracious consumers of a wide range of organic materials ranging from fruits and vegetables to animal remains, and manure. Thanks to this ability and considering the larval high protein and lipid content, BSF larvae are a useful additive in animal feeds and biodiesel production. Unfortunately, the feasibility of using the black soldier fly as a tool for waste valorization and feed production has primarily been investigated at the benchtop scale. Thus, mobilization of current practices to an industrial scale is challenging because scaling up from small laboratory studies to large industrial studies is not necessarily linear. The goal of this study was to demonstrate the ability of the BSF to recycle organic waste at an industrial scale. To accomplish this goal, three organic waste streams were used (e.g., apples, bananas, and spent grain from a brewery) to test six diet treatments (1) apple, (2) banana, (3) spent grain, (4) apple and banana, (5) apple and spent grain, and (6) banana and spent grain. Working at scale of 10,000 BSF larvae life history traits, waste valorization, protein and lipid profiles were measured for each diet treatment. Differences were recorded across all variables, except substrate conversion, for larvae fed on fruit and spent grain (alone or with fruit). Growth rate significantly differed across treatments; larvae reared on spent grain grew twice as fast as those fed apples alone, but those reared on the apple and spent grain mixture produced twice as much insect biomass. However, it should be noted that larvae resulting from the apple diet contained 50% more fat than larvae fed the fruit and spent grain mixtures. Commonly-available organic wastes were successfully used at an industrial scale to produce BSF larvae that have the potential to substitute other sources of protein and lipids in different industrial applications. Industrialization efforts are encouraged to assess these impacts when integrating diverse ingredients into larval diets as a means to more precisely predict output, such as larval development time and final larval biomass