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The larvae of the black soldier fly (BSF), Hermetia illucens (Linnaeus, 1758) (Diptera: Stratiomyidae), are considered an efficient system for the bioconversion of organic waste into usable products, such as insect protein for animal feed and bioactive molecules. Despite the great interest toward H. illucens and its biotechnological applications, information on the biology of this insect is still scarce. In particular, no data on the structural and functional properties of the digestive system of the adult insect are available and it is a common belief that the fly does not eat. In the present work, we therefore investigate the remodeling process of the BSF larval midgut during metamorphosis, analyze the morphofunctional properties of the adult midgut, evaluate if the fly is able to ingest and digest food and assess whether the feeding supply influences the adult performances. Our results show that the larval midgut of H. illucens is removed during metamorphosis and a new pupal-adult epithelium, characterized by peculiar features compared to the larval organ, is formed by proliferation and differentiation of midgut stem cells. Moreover, our experiments indicate that the adult insect possesses a functional digestive system and that food administration affects the longevity of the fly. These data not only demonstrate that the adult BSF is able to eat but also open up the possibility to manipulate the feeding substrate of the fly to improve its performances in mass rearing procedures.

Background In the last few years, considerable attention has been focused on the plastic-degrading capability of insects and their gut microbiota in order to develop novel, effective, and green strategies for plastic waste management. Although many analyses based on 16S rRNA gene sequencing are available, an in-depth analysis of the insect gut microbiome to identify genes with plastic-degrading potential is still lacking. Results In the present work, we aim to fill this gap using Black Soldier Fly (BSF) as insect model. BSF larvae have proven capability to efficiently bioconvert a wide variety of organic wastes but, surprisingly, have never been considered for plastic degradation. BSF larvae were reared on two widely used plastic polymers and shotgun metagenomics was exploited to evaluate if and how plastic-containing diets affect composition and functions of the gut microbial community. The high-definition picture of the BSF gut microbiome gave access for the first time to the genomes of culturable and unculturable microorganisms in the gut of insects reared on plastics and revealed that (i) plastics significantly shaped bacterial composition at species and strain level, and (ii) functions that trigger the degradation of the polymer chains, i.e., DyP-type peroxidases, multicopper oxidases, and alkane monooxygenases, were highly enriched in the metagenomes upon exposure to plastics, consistently with the evidences obtained by scanning electron microscopy and 1 H nuclear magnetic resonance analyses on plastics. Conclusions In addition to highlighting that the astonishing plasticity of the microbiota composition of BSF larvae is associated with functional shifts in the insect microbiome, the present work sets the stage for exploiting BSF larvae as “bioincubators” to isolate microbial strains and enzymes for the development of innovative plastic biodegradation strategies. However, most importantly, the larvae constitute a source of enzymes to be evolved and valorized by pioneering synthetic biology approaches. 5errh-AhYWWkX5YWmGozvJ Video Abstract

The calcified tissues of vertebrates are essentially represented by bone, cartilage, dentin and calcified tendons. In all these tissues a major hallmark of mineralization is the deposition of the inorganic phase on a pre-existing collagen template, but evident differences exist among these materials and the molecular details of the process are still incompletely understood. In this study, the ultrastructural aspects of the mineral phase of these tissues were investigated by means of high-resolution scanning electron microscopy (HR-SEM) after low-temperature thermal deproteination, a technique allowing a direct, unrestricted visualization of the mineral component. Each tissue showed distinctive features. In most cases, calcification proceeds in a discontinuous way through the formation of clumps or clusters of mineralized tissue; in all cases, except cartilage, the mineral phase shows an evident relationship with the layout and/or the D-period of the collagen fibrils. Our results highlight the peculiar aspect of the mineralization process in the cartilage with respect to the other tissues, all of them containing collagen type I instead of type II, and suggest that a different molecular mechanism may be at work. It is still unclear whether and how this may be related to the content, exclusive of cartilage, of collagen type II. The identification of the tissue-specific features exhibited by cartilage versus those shared by all the other three tissues, although from different species, requires further research on physiological calcification.

In the Chondrichthyes Raja asterias and Myliobatis aquila and in the Teleost Sparus aurata, the appendicular skeleton of the pectoral fins (including the calcified structures of the mouth in M. aquila) was investigated to find out how the specific skeletal segments were formed and stiffened over the course of evolution, not only with regard to the adaptation of the ontogenesis of the cartilage “anlagen” to the mechanical requirements of locomotion in the water column, but also to the specific feeding habits (durophagy) of M. aquila. The morphology of the pectoral fins of the three species showed a different layout, characterized by the geometry of the basic units (aligned tesserae and calcified radial columns), which provide varied flexibility of the pectoral fins, suggesting an adaptation to the “pelagic” and “benthic” locomotion patterns in the environment where the species live. The morphology of the calcified structures in the mouth of M. aquila showed the presence of two different masticatory systems: the first (external) with the rows of teeth resting on the maxillary and mandibular arches, and the second (internal, in the oral cavity) with the symphyseal plates specialized for durophagy. Chemical–physical analyses revealed that the calcified cartilage matrix of the Chondrichthyes fin rays, teeth and durophagy plates is stiffened by the same Ca3(PO4)2 mineral phase deposed in the organic matrix of the Teleost S. aurata fins (with the characteristic SEM morphological texture of calcified bone matrix). The hitherto unknown presence of two different chewing systems in M. aquila documents an evolutionary adaptation to nutritional requirements that can be explained by two hypotheses: the coexistence of two functioning systems in current specimens, allowing for the ingestion of harder and softer prey (or plant food), or the persistence of a rudimentary dentition that is no longer used (vestigial dentition). Furthermore, the texture of the calcified matrix in teleost fishes, as observed by scanning electron microscopy, may indicate a bone-like organic matrix substrate, similar to that found in endochondral ossification.

The increase in the world population and consequent rise in food demand have led to the extensive use of chemical pesticides, causing environmental and health concerns. In response, biological control methods, particularly those involving microbial agents, have emerged as sustainable alternatives within integrated pest management. This study highlights the potential of Lysinibacillus fusiformis as a biocontrol agent against the dipteran Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), a pest responsible for damaging soft-skinned fruits. Experimental treatments using vegetative cells, spores, and secondary metabolites of L. fusiformis on D. suzukii larvae demonstrated significant larvicidal effects, accompanied by observable changes in gut morphology under microscopy. Moreover, preliminary immunological assays showed the interference of this bacterium with the host immune system. All the results indicate the suitability of L. fusiformis for its possible integration into sustainable agricultural practices, although additional research is required to understand its applicability in the field.

Introduction Traditional orthodontic diagnostics rely significantly on lateral cephalometric radiographs, posing health risks due to ionising radiation, particularly in paediatric patients. Artificial intelligence (AI) represents a promising alternative by enabling predictions of cephalometric parameters from non-radiographic clinical data. This study evaluates the accuracy and clinical utility of CEPHCLINIC, an open-source AI software designed to predict conventional cephalometric measurements using clinical photographs and intraoral 3D scans, thus adhering to radiation protection principles. Materials and methods The dataset comprised 1255 subjects from the American Association of Orthodontists Foundation (AAOF) craniofacial collection, encompassing demographic and clinical variables (age, gender, overbite, overjet, facial dimensions). This dataset was randomly divided into training (80%, n  = 1004) and validation (20%, n  = 251) subsets. Additionally, an independent external test set of 51 untreated orthodontic cases was employed for rigorous evaluation. Input variables for model training included clinical parameters derived from photographs (WebCeph software) and intraoral scans (iTero scanner, MeshMixer software). Supervised predictive regression models, including ExtraTreesRegressor, CatBoostRegressor, and Support Vector Regression, were optimised through GridSearchCV and validated using repeated random subsampling. Predictive accuracy was assessed statistically using Root Mean Squared Error (RMSE), Mean Absolute Error (MAE), Spearman correlation coefficients, R-squared values, and paired t-tests. Results The ExtraTreesRegressor demonstrated superior performance across multiple cephalometric parameters, achieving notably low RMSE values in the independent test set for ANB (2.772°) and NP2PA (2.317 mm). However, parameters like COPAD exhibited higher prediction errors (RMSE 12.121 mm). Spearman correlation analysis indicated strong prediction consistency for COPOD (0.850), moderate for U1SNA (0.548), and poor predictability for NP2PO (-0.052). Despite statistically significant biases observed in predictions for some parameters (paired t-test, p  < 0.05), overall predictive accuracy was clinically acceptable, emphasising parameters such as COPOD, COPAD, ANB, and U1SNA as particularly reliable. Conclusions The CEPHCLINIC software reliably predicts key cephalometric measurements from non-radiographic clinical data, significantly reducing radiation exposure risks. Despite promising performance, model refinement, dataset expansion with broader demographic representation, and integration with digital orthodontic technologies are essential for enhancing precision, clinical reliability, and global applicability.

Investigating morphological and molecular mechanisms that plants adopt in response to artificial biophilic lighting is crucial for implementing biophilic approaches in indoor environments. Also, studying the essential oils (EOs) composition in aromatic plants can help unveil the light influence on plant metabolism and open new investigative routes devoted to producing valuable molecules for human health and commercial applications. We assessed the growth performance and the EOs composition of Mentha x piperita and Ocimum basilicum grown under an innovative artificial biophilic lighting system (CoeLux ® ), that enables the simulation of natural sunlight with a realistic sun perception, and compared it to high-pressure sodium lamps (control) We found that plants grown under the CoeLux ® light type experienced a general suppression of both above and belowground biomass, a high leaf area, and a lower leaf thickness, which might be related to the shade avoidance syndrome. The secondary metabolites composition in the plants’ essential oils was scarcely affected by both light intensity and spectral composition of the CoeLux ® light type, as similarities above 80% were observed with respect to the control light treatments and within both plant species. The major differences were detected with respect to the EOs extracted from plants grown under natural sunlight (52% similarity in M. piperita and 75% in O. basilicum ). Overall, it can be speculated that the growth of these two aromatic plants under the CoeLux ® lighting systems is a feasible strategy to improve biophilic approaches in closed environments that include both plants and artificial sunlight. Among the two plant species analyzed, O. basilicum showed an overall better performance in terms of both morphological traits and essential oil composition. To increase biomass production and enhance the EOs quality (e.g., higher menthol concentrations), further studies should focus on technical solutions to raise the light intensity irradiating plants during their growth under the CoeLux ® lighting systems.

The innate immune response represents a first-line defense against pathogen infection that has been widely conserved throughout evolution. Using the invertebrate (Annelida, Hirudinea) as an experimental model, we show here that the RNASET2 ribonuclease is directly involved in the immune response against Gram-positive bacteria. Injection of lipoteichoic acid (LTA), a key component of Gram-positive bacteria cell wall, into the leech body wall induced a massive migration of granulocytes and macrophages expressing TLR2 (the key receptor involved in the response to Gram-positive bacteria) toward the challenged/inoculated area. We hypothesized that the endogenous leech RNASET2 protein ( RNASET2) might be involved in the antimicrobial response, as already described for other vertebrate ribonucleases, such as RNase3 and RNase7. In support of our hypothesis, RNASET2 was mainly localized in the granules of granulocytes, and its release in the extracellular matrix triggered the recruitment of macrophages toward the area stimulated with LTA. The activity of RNASET2 was also evaluated on living cells by means of light, transmission, and scanning electron microscopy analysis. RNASET2 injection triggered the formation of clumps following a direct interaction with the bacterial cell wall, as demonstrated by immunogold assay. Taken together, our data support the notion that, during the early phase of leech immune response, granulocyte-released RNASET2 triggers bacterial clumps formation and, at the same time, actively recruits phagocytic macrophages in order to elicit a rapid and effective eradication of the infecting microorganisms from inoculated area.

The science of dental tissue grafting is evolving, with an increased understanding of factors influencing graft behavior. Despite the widespread clinical use of soft tissue grafts, the histological characteristics of different gingival harvesting sites are still underexplored. This study aimed to fill this gap by analyzing 50 tissue samples harvested from 25 patients across three sites: the hard palate, maxillary tuberosity, and palatal rugae. Each sample underwent thorough histological and histomorphometric analysis. Conventional statistical analysis was performed using SPSS, while predictive modeling was conducted with RapidMiner Studio. The study identified significant histological differences among the graft sites, with notable variations in total graft height, epithelial height, and interdigitation perimeter. These findings underscore the importance of donor site selection in influencing graft success. Pair plots and principal component analysis (PCA) further highlighted the distinct histological features of each tissue type. The random forest classifier identified total graft height, epithelial height, and perimeter as the most influential factors in predicting graft site behavior. This study offers valuable insights into the histological characteristics of soft tissue grafts, potentially leading to more predictable clinical outcomes.

This study aimed to investigate the remineralization processes of dental enamel via scanning electron microscopy and observe the changes induced in the microstructure by oral hygiene products. More specifically, the effectiveness of Mentadent Professional Resilience was analyzed for its ability to demineralize erosion-affected enamel surfaces. This involved the sectioning of some teeth to preserve enamel integrity, followed by dehydration and preparation for SEM analysis. SEM observations were made at various magnifications to detect differences in enamel morphology after treatment with the product. These observations provide valuable insights into the mechanisms of action of dental care products and their potential to protect enamel. The study makes a contribution to our understanding of remineralization processes and describes the importance of microscopic analysis for evaluating and developing effective dental products.