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Plants generate energy flows through natural food webs, driven by competition for resources among organisms, which are part of a complex network of multitrophic interactions. Here, we demonstrate that the interaction between tomato plants and a phytophagous insect is driven by a hidden interplay between their respective microbiotas. Tomato plants colonized by the soil fungus Trichoderma afroharzianum, a beneficial microorganism widely used in agriculture as a biocontrol agent, negatively affects the development and survival of the lepidopteran pest Spodoptera littoralis by altering the larval gut microbiota and its nutritional support to the host. Indeed, experiments aimed to restore the functional microbial community in the gut allow a complete rescue. Our results shed light on a novel role played by a soil microorganism in the modulation of plant-insect interaction, setting the stage for a more comprehensive analysis of the impact that biocontrol agents may have on ecological sustainability of agricultural systems.
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Microbioma Gastrointestinal , Microbiota , Solanum lycopersicum , Animales , Suelo , Insectos , AgriculturaRESUMEN
Modulation of nutrient digestion and absorption is one of the post-ingestion mechanisms that guarantees the best exploitation of food resources, even when they are nutritionally poor or unbalanced, and plays a pivotal role in generalist feeders, which experience an extreme variability in diet composition. Among insects, the larvae of black soldier fly (BSF), Hermetia illucens, can grow on a wide range of feeding substrates with different nutrient content, suggesting that they can set in motion post-ingestion processes to match their nutritional requirements. In the present study we address this issue by investigating how the BSF larval midgut adapts to diets with different nutrient content. Two rearing substrates were compared: a nutritionally balanced diet for dipteran larvae and a nutritionally poor diet that mimics fruit and vegetable waste. Our data show that larval growth performance is only moderately affected by the nutritionally poor diet, while differences in the activity of digestive enzymes, midgut cell morphology, and accumulation of long-term storage molecules can be observed, indicating that diet-dependent adaptation processes in the midgut ensure the exploitation of poor substrates. Midgut transcriptome analysis of larvae reared on the two substrates showed that genes with important functions in digestion and absorption are differentially expressed, confirming the adaptability of this organ.
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Dieta , Dípteros/fisiología , Adaptación Fisiológica , Alimentación Animal/análisis , Animales , Peso Corporal , Carbohidratos de la Dieta/análisis , Carbohidratos de la Dieta/farmacocinética , Proteínas en la Dieta/análisis , Proteínas en la Dieta/farmacocinética , Dípteros/genética , Dípteros/crecimiento & desarrollo , Frutas , Regulación de la Expresión Génica , Ontología de Genes , Concentración de Iones de Hidrógeno , Absorción Intestinal , Mucosa Intestinal/metabolismo , Intestinos/fisiología , Larva , Nutrientes/análisis , Nutrientes/farmacocinética , Pupa , RNA-Seq , Transcriptoma , VerdurasRESUMEN
The larva of the black soldier fly (Hermetia illucens) has emerged as an efficient system for the bioconversion of organic waste. Although many research efforts are devoted to the optimization of rearing conditions to increase the yield of the bioconversion process, microbiological aspects related to this insect are still neglected. Here, we describe the microbiota of the midgut of H. illucens larvae, showing the effect of different diets and midgut regions in shaping microbial load and diversity. The bacterial communities residing in the three parts of the midgut, characterized by remarkable changes in luminal pH values, differed in terms of bacterial numbers and microbiota composition. The microbiota of the anterior part of the midgut showed the highest diversity, which gradually decreased along the midgut, whereas bacterial load had an opposite trend, being maximal in the posterior region. The results also showed that the influence of the microbial content of ingested food was limited to the anterior part of the midgut, and that the feeding activity of H. illucens larvae did not significantly affect the microbiota of the substrate. Moreover, a high protein content compared to other macronutrients in the feeding substrate seemed to favor midgut dysbiosis. The overall data indicate the importance of taking into account the presence of different midgut structural and functional domains, as well as the substrate microbiota, in any further study that aims at clarifying microbiological aspects concerning H. illucens larval midgut.IMPORTANCE The demand for food of animal origin is expected to increase by 2050. Since traditional protein sources for monogastric diets are failing to meet the increasing demand for additional feed production, there is an urgent need to find alternative protein sources. The larvae of Hermetia illucens emerge as efficient converters of low-quality biomass into nutritionally valuable proteins. Many studies have been performed to optimize H. illucens mass rearing on a number of organic substrates and to quantitatively and qualitatively maximize the biomass yield. On the contrary, although the insect microbiota can be fundamental for bioconversion processes and its characterization is mandatory also for safety aspects, this topic is largely overlooked. Here, we provide an in-depth study of the microbiota of H. illucens larval midgut, taking into account pivotal aspects, such as the midgut spatial and functional regionalization, as well as microbiota and nutrient composition of the feeding substrate.
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Dípteros/microbiología , Microbioma Gastrointestinal , Alimentación Animal/análisis , Animales , Dieta , Sistema Digestivo/efectos de los fármacos , Sistema Digestivo/microbiología , Dípteros/crecimiento & desarrollo , Dípteros/fisiología , Microbioma Gastrointestinal/efectos de los fármacos , Larva/crecimiento & desarrollo , Larva/microbiología , Larva/fisiologíaRESUMEN
The insect midgut epithelium represents an interface between the internal and the external environment and it is the almost unique epithelial tissue by which these arthropods acquire nutrients. This epithelium is indeed able to produce digestive enzymes and to support vectorial transport of small organic nutrients, ions, and water. Moreover, it plays a key role in the defense against pathogenic microorganisms and in shaping gut microbiota. Another important midgut function is the ability to produce signaling molecules that regulate its own physiology and the activity of other organs. The two main mature cell types present in the midgut of all insects, i.e., columnar and endocrine cells, are responsible for these functions. In addition, stem cells, located at the base of the midgut epithelium, ensure the growth and renewal of the midgut during development and after injury. In insects belonging to specific orders, midgut physiology is deeply conditioned by the presence of unique cell types, i.e., goblet and copper cells, which confer peculiar features to this organ. This review reports current knowledge on the cells that form the insect midgut epithelium, focusing attention on their morphological and functional features. Notwithstanding the apparent structural simplicity of this organ, the properties of the cells make the midgut a key player in insect development and homeostasis.
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Sistema Digestivo/ultraestructura , Endodermo/ultraestructura , Insectos/anatomía & histología , AnimalesRESUMEN
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.
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Sistema Digestivo/metabolismo , Dípteros/fisiología , Proteínas de Insectos/metabolismo , Larva/fisiología , Pupa/fisiología , Alimentación Animal , Animales , Metamorfosis BiológicaRESUMEN
Bacillus thuringiensis is a widely used bacterial entomopathogen producing insecticidal toxins, some of which are expressed in insect-resistant transgenic crops. Surprisingly, the killing mechanism of B. thuringiensis remains controversial. In particular, the importance of the septicemia induced by the host midgut microbiota is still debated as a result of the lack of experimental evidence obtained without drastic manipulation of the midgut and its content. Here this key issue is addressed by RNAi-mediated silencing of an immune gene in a lepidopteran host Spodoptera littoralis, leaving the midgut microbiota unaltered. The resulting cellular immunosuppression was characterized by a reduced nodulation response, which was associated with a significant enhancement of host larvae mortality triggered by B. thuringiensis and a Cry toxin. This was determined by an uncontrolled proliferation of midgut bacteria, after entering the body cavity through toxin-induced epithelial lesions. Consequently, the hemolymphatic microbiota dramatically changed upon treatment with Cry1Ca toxin, showing a remarkable predominance of Serratia and Clostridium species, which switched from asymptomatic gut symbionts to hemocoelic pathogens. These experimental results demonstrate the important contribution of host enteric flora in B. thuringiensis-killing activity and provide a sound foundation for developing new insect control strategies aimed at enhancing the impact of biocontrol agents by reducing the immunocompetence of the host.
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Bacillus thuringiensis/patogenicidad , Proteínas Bacterianas/biosíntesis , Endotoxinas/biosíntesis , Proteínas Hemolisinas/biosíntesis , Proteínas de Insectos/antagonistas & inhibidores , Microbiota/inmunología , Control Biológico de Vectores/métodos , Spodoptera/inmunología , Animales , Bacillus thuringiensis/crecimiento & desarrollo , Toxinas de Bacillus thuringiensis , Clostridium/crecimiento & desarrollo , Clostridium/patogenicidad , Productos Agrícolas/parasitología , Regulación de la Expresión Génica , Hemocitos/inmunología , Hemocitos/microbiología , Inmunidad Innata , Terapia de Inmunosupresión , Proteínas de Insectos/genética , Proteínas de Insectos/inmunología , Intestinos/inmunología , Intestinos/microbiología , Larva/genética , Larva/inmunología , Larva/microbiología , Interferencia de ARN , ARN Bicatenario/genética , ARN Bicatenario/metabolismo , Serratia/crecimiento & desarrollo , Serratia/patogenicidad , Spodoptera/genética , Spodoptera/microbiologíaRESUMEN
The midgut represents the middle part of the alimentary canal and is responsible for nutrient digestion and absorption in insect larva. Despite the growing interest in this organ for different purposes, such as studies on morphogenesis and differentiation, stem cell biology, cell death processes and transport mechanisms, basic information on midgut development is still lacking for a large proportion of insect species. Undoubtedly, this lack of data could hinder the full exploitation of practical applications that involve midgut as their primary target. This may represent in particular a significant problem for Lepidoptera, an insect order that includes some of the most important species of high economic importance. With the aim of overcoming this fragmentation of knowledge, we performed a detailed morphofunctional analysis of the midgut of the silkworm, Bombyx mori, a representative model among Lepidoptera, during its development from the larval up to the adult stage, focusing attention on stem cells. Our data demonstrate stem cell proliferation and differentiation, not only in the larval midgut but also in the pupal and adult midgut epithelium. Moreover, we present evidence for a complex trophic relationship between the dying larval epithelium and the new adult one, which is established during metamorphosis. This study, besides representing the first morphological and functional characterization of the changes that occur in the midgut of a lepidopteron during the transition from the larva to the moth, provides a detailed analysis of the midgut of the adult insect, a stage that has been neglected up to now.
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Bombyx/citología , Bombyx/crecimiento & desarrollo , Epitelio/crecimiento & desarrollo , Células Madre/citología , Animales , Muerte Celular , Diferenciación Celular , Proliferación Celular , Células Epiteliales/citología , Larva/citología , Larva/crecimiento & desarrollo , Metamorfosis BiológicaRESUMEN
7-Deoxy-uniflorine A (6), synthesized ex novo with a straightforward and simple strategy, and the analogues 4, 5 and 7, were evaluated as potential inhibitors of insect trehalase from Chironomus riparius and Spodoptera littoralis. All the compounds were tested against porcine trehalase as the mammalian counterpart and α-amylase from human saliva as a relevant glucolytic enzyme. The aim of this work is the identification of the simplest pyrrolizidine structure necessary to impart selective insect trehalase inhibition, in order to identify new specific inhibitors that can be easily synthesized compared to our previous reports with the potential to act as non-toxic insecticides and/or fungicides. All the derivatives 47 proved to be active (from low micromolar to high nanomolar range activity) towards insect trehalases, while no activity was observed against α-amylase. In particular, the natural compound uniflorine A and its 7-deoxy analogue were found to selectively inhibit insect trehalases, as they are inactive towards the mammalian enzyme. The effect of compound 6 was also analyzed in preliminary in vivo experiments. These new findings allow the identification of natural uniflorine A and its 7-deoxy analogue as the most promising inhibitors among a series of pyrrolizidine derivatives for future development in the agrochemical field, and the investigation also outlined the importance of the stereochemistry at C-6 of pyrrolizidine nucleus to confer such enzyme specificity.
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Inhibidores Enzimáticos/química , Indolizinas/química , Proteínas de Insectos/antagonistas & inhibidores , Insecticidas/química , Alcaloides de Pirrolicidina/química , Trehalasa/antagonistas & inhibidores , Animales , Chironomidae/química , Chironomidae/efectos de los fármacos , Chironomidae/enzimología , Pruebas de Enzimas , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Humanos , Indolizinas/síntesis química , Indolizinas/farmacología , Proteínas de Insectos/química , Insecticidas/síntesis química , Insecticidas/farmacología , Cinética , Larva/química , Larva/efectos de los fármacos , Larva/enzimología , Alcaloides de Pirrolicidina/síntesis química , Alcaloides de Pirrolicidina/farmacología , Especificidad de la Especie , Spodoptera/química , Spodoptera/efectos de los fármacos , Spodoptera/enzimología , Porcinos , Trehalasa/química , alfa-Amilasas/químicaRESUMEN
PURPOSE: Myiases are infestations of human and animal tissues by fly larvae. These conditions are widespread in tropical countries and travelers in those areas are at risk of becoming infested. Although Cordylobia anthropophaga (Blanchard & Berenger-Feraud, 1872) is one of the most common myiasis-causing species, few high-quality images and molecular sequences are available for this fly. We present a case of C. anthropophaga infestation in an Italian patient returning from Senegal, with the aim of increasing both visual and molecular data for this species. METHODS: After removal, the larva was determined following standardized morphological keys and photographed under a digital microscope. Molecular characterization of the Cytochrome c oxidase subunit I (COI) was performed using universal primers. RESULTS: The general appearance, the structural organization of the cephalic region, of the cephaloskeleton, and of the posterior tracheal spiracles suggested that the causative agent of the myiasis was a third instar larva of C. anthropophaga. The morphological data are further supported by the molecular data: the COI sequence showed high levels of identity with the already published verified COI sequences of C. anthropophaga. CONCLUSION: We provide high-quality morphological and molecular data useful for the identification of larvae of C. anthropophaga. We highlight that myiasis might be common in Senegal and better data about its prevalence in travelers and in the endemic countries are needed to understand the burden of this condition.
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Calliphoridae , Miasis , Viaje , Animales , Humanos , Masculino , Complejo IV de Transporte de Electrones/genética , Italia , Larva , Miasis/parasitología , Senegal , Persona de Mediana EdadRESUMEN
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 Society of Chemical Industry.
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Bacillus thuringiensis , Quitinasas , Larva , Mariposas Nocturnas , Control Biológico de Vectores , Quitinasas/metabolismo , Animales , Mariposas Nocturnas/efectos de los fármacos , Mariposas Nocturnas/crecimiento & desarrollo , Larva/crecimiento & desarrollo , Larva/efectos de los fármacos , Hypocreales/enzimología , Proteínas Fúngicas/metabolismo , Agentes de Control Biológico/farmacologíaRESUMEN
Densoviruses are insect parvoviruses that are orally infectious for Lepidoptera. To assess the mechanisms underlying their specificity and their virulence, we investigated the role of eight candidate residues in the densovirus capsid. We showed that the substitutions of four amino acids were associated with decreased virulence due to a decreased ability to cross the host midgut epithelium, without an effect on viral replication in other tissues.
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Proteínas de la Cápside/química , Proteínas de la Cápside/metabolismo , Densovirus/fisiología , Densovirus/patogenicidad , Spodoptera/virología , Tropismo Viral , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Proteínas de la Cápside/genética , Densovirus/química , Densovirus/genética , Intestinos/virología , Modelos Moleculares , Datos de Secuencia Molecular , Virulencia , Replicación ViralRESUMEN
In insects, the cell-mediated immune response involves an active role of hemocytes in phagocytosis, nodulation, and encapsulation. Although these processes have been well documented in multiple species belonging to different insect orders, information concerning the immune response, particularly the hemocyte types and their specific function in the black soldier fly Hermetia illucens, is still limited. This is a serious gap in knowledge given the high economic relevance of H. illucens larvae in waste management strategies and considering that the saprophagous feeding habits of this dipteran species have likely shaped its immune system to efficiently respond to infections. The present study represents the first detailed characterization of black soldier fly hemocytes and provides new insights into the cell-mediated immune response of this insect. In particular, in addition to prohemocytes, we identified five hemocyte types that mount the immune response in the larva, and analyzed their behavior, role, and morphofunctional changes in response to bacterial infection and injection of chromatographic beads. Our results demonstrate that the circulating phagocytes in black soldier fly larvae are plasmatocytes. These cells also take part in nodulation and encapsulation with granulocytes and lamellocyte-like cells, developing a starting core for nodule/capsule formation to remove/encapsulate large bacterial aggregates/pathogens from the hemolymph, respectively. These processes are supported by the release of melanin precursors from crystal cells and likely by mobilizing nutrient reserves in newly circulating adipohemocytes, which could thus trophically support other hemocytes during the immune response. Finally, the regulation of the cell-mediated immune response by eicosanoids was investigated.
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Dípteros , Hemocitos , Animales , Larva/fisiología , Dípteros/fisiología , Fagocitosis/fisiología , HemolinfaRESUMEN
Some insect species have gained attention as efficient bioconverters of low-value organic substrates (i.e., residual streams) into high-value biomass. Black soldier fly (BSF) (Hermetia illucens) larvae are particularly interesting for bioconversion due to their ability to grow on a wide range of substrates, including low-value industrial residual streams. This is in part due to the plasticity of the gut microbiota of polyphagous insects, like BSF. Gut microbiota composition varies depending on rearing substrates, via a mechanism that might support the recruitment of microorganisms that facilitate digestion of a specific substrate. At the same time, specific microbial genera do persist on different substrates via unknown mechanisms. This study aimed to offer insights on this microbial plasticity by investigating how the composition of the bacterial community present in the gut of BSF larvae responds to two industrial residual streams: swill (a mixture of catering and supermarket leftovers) and distiller's dried grains with solubles. The bacterial biota composition of substrates, whole larvae at the beginning of the rearing period and at harvest, rearing residues, and larval gut regions were investigated through 16S rRNA gene sequencing. It was observed that both substrate and insect development influenced the bacterial composition of the whole larvae. Zooming in on the gut regions, there was a clear shift in community composition from a higher to a lower diversity between the anterior/middle midgut and the posterior midgut/hindgut, indicating a selective pressure occurring in the middle midgut region. Additionally, the abundance of the bacterial biota was always high in the hindgut, while its diversity was relatively low. Even more, the bacterial community in the hindgut was found to be relatively more conserved over the different substrates, harboring members of the BSF core microbiota. We postulate a potential role of the hindgut as a reservoir for insect-associated microbes. This warrants further research on that underexplored region of the intestinal tract. Overall, these findings contribute to our understanding of the bacterial biota structure and dynamics along the intestinal tract, which can aid microbiome engineering efforts to enhance larval performance on (industrial) residual streams.
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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 1H 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. Video Abstract.
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Dípteros , Microbioma Gastrointestinal , Animales , Larva , Microbioma Gastrointestinal/genética , Plásticos , ARN Ribosómico 16S/genéticaRESUMEN
Although several features of apoptosis and autophagy have been reported in the larval organs of Lepidoptera during metamorphosis, solid experimental evidence for autophagy is still lacking. Moreover, the role of the two processes and the nature of their relationship are still cryptic. In this study, we perform a cellular, biochemical and molecular analysis of the degeneration process that occurs in the larval midgut of Bombyx mori during larval-adult transformation, with the aim to analyze autophagy and apoptosis in cells that die under physiological conditions. We demonstrate that larval midgut degradation is due to the concerted action of the two mechanisms, which occur at different times and have different functions. Autophagy is activated from the wandering stage and reaches a high level of activity during the spinning and prepupal stages, as demonstrated by specific autophagic markers. Our data show that the process of autophagy can recycle molecules from the degenerating cells and supply nutrients to the animal during the non-feeding period. Apoptosis intervenes later. In fact, although genes encoding caspases are transcribed at the end of the larval period, the activity of these proteases is not appreciable until the second day of spinning and apoptotic features are observable from prepupal phase. The abundance of apoptotic features during the pupal phase, when the majority of the cells die, indicates that apoptosis is actually responsible for cell death and for the disappearance of larval midgut cells.
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Apoptosis/fisiología , Autofagia/fisiología , Bombyx/crecimiento & desarrollo , Metamorfosis Biológica/fisiología , Animales , Autofagia/genética , Bombyx/citología , Bombyx/metabolismo , Caspasas/metabolismo , Sistema Digestivo/anatomía & histología , Sistema Digestivo/metabolismo , Larva/citología , Larva/metabolismo , Larva/ultraestructura , Pupa/metabolismoRESUMEN
The various subjects covered in the present Special Issue "Silkworm and Silk: Traditional and Innovative Applications" demonstrate how sericulture, a practice deeply rooted in human history, can act as a bridge to bring together an exceptionally wide range of scientific and technical expertise in both conventional topics and cutting-edge technologies [...].
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In mountain ecosystems, climate change can cause spatiotemporal shifts, impacting the composition of communities and altering fundamental biotic interactions, such as those involving flower-visiting arthropods. On of the main problems in assessing the effects of climate change on arthropods in these environments is the lack of baseline data. In particular, the arthropod communities on early flowering high-altitude plants are poorly investigated, although the early season is a critical moment for possible mismatches. In this study, we characterised the flower-visiting arthropod community on the early flowering high-altitude Alpine plant, Androsace brevis (Primulaceae). In addition, we tested the effect of abiotic factors (temperature and wind speed) and other variables (time, i.e., hour of the day, and number of flowers per plant) on the occurrence, abundance, and diversity of this community. A. brevis is a vulnerable endemic species growing in the Central Alps above 2000 m asl and flowering for a very short period immediately after snowmelt, thus representing a possible focal plant for arthropods in this particular moment of the season. Diptera and Hymenoptera were the main flower visitors, and three major features of the community emerged: an evident predominance of anthomyiid flies among Diptera, a rare presence of bees, and a relevant share of parasitoid wasps. Temperature and time (hour of the day), but not wind speed and number of flowers per plant, affected the flower visitors' activity. Our study contributes to (1) defining the composition of high-altitude Alpine flower-visiting arthropod communities in the early season, (2) establishing how these communities are affected by environmental variables, and (3) setting the stage for future evaluation of climate change effects on flower-visiting arthropods in high-altitude environments in the early season.
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In insects, a complex and effective immune system that can be rapidly activated by a plethora of stimuli has evolved. Although the main cellular and humoral mechanisms and their activation pathways are highly conserved across insects, the timing and the efficacy of triggered immune responses can differ among different species. In this scenario, an insect deserving particular attention is the black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae). Indeed, BSF larvae can be reared on a wide range of decaying organic substrates and, thanks to their high protein and lipid content, they represent a valuable source of macromolecules useful for different applications (e.g., production of feedstuff, bioplastics, and biodiesel), thus contributing to the development of circular economy supply chains for waste valorization. However, decaying substrates bring the larvae into contact with different potential pathogens that can challenge their health status and growth. Although these life strategies have presumably contributed to shape the evolution of a sophisticated and efficient immune system in this dipteran, knowledge about its functional features is still fragmentary. In the present study, we investigated the processes underpinning the immune response to bacteria in H. illucens larvae and characterized their reaction times. Our data demonstrate that the cellular and humoral responses in this insect show different kinetics: phagocytosis and encapsulation are rapidly triggered after the immune challenge, while the humoral components intervene later. Moreover, although both Gram-positive and Gram-negative bacteria are completely removed from the insect body within a few hours after injection, Gram-positive bacteria persist in the hemolymph longer than do Gram-negative bacteria. Finally, the activity of two key actors of the humoral response, i.e., lysozyme and phenoloxidase, show unusual dynamics as compared to other insects. This study represents the first detailed characterization of the immune response to bacteria of H. illucens larvae, expanding knowledge on the defense mechanisms of this insect among Diptera. This information is a prerequisite to manipulating the larval immune response by nutritional and environmental factors to increase resistance to pathogens and optimize health status during mass rearing.
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Inmunidad/inmunología , Larva/inmunología , Larva/microbiología , Simuliidae/inmunología , Simuliidae/microbiología , Animales , Bacterias/inmunologíaRESUMEN
The aim of this work was to develop processing methods that safeguard the quality and antimicrobial properties of H. illucens and B. mori oils. We adopted a vegetable diet for both insects: leftover vegetables and fruit for H. illucens and mulberry leaves for B. mori. First, alternative techniques to obtain a good oil extraction yield from the dried biomass of H. illucens larvae were tested. Traditional pressing resulted to be the best system to maximize the oil yield and it was successfully applied to B. mori pupae. Oil quality resulted comparable to that obtained with other extraction methods described in the literature. In the case of B. mori pupae, different treatments and preservation periods were investigated to evaluate their influence on the oil composition and quality. Interestingly, agar diffusion assays demonstrated the sensitivity of Gram-positive Bacillus subtilis and Staphylococcus aureus to H. illucens and B. mori derived oils, whereas the growth of Gram-negative Pseudomonas aeruginosa and Escherichia coli was not affected. This study confirms that fat and other active compounds of the oil extracted by hot pressing could represent effective antimicrobials against bacteria, a relevant result if we consider that they are by-products of the protein extraction process in the feed industry.
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The larvae of the black soldier fly (BSF) Hermetia illucens are increasingly being used for waste management purposes given their ability to grow on a wide range of organic decaying materials. Although significant efforts have been spent to improve the mass rearing of BSF larvae on specific substrates and their bioconversion capability, little is known about the biology of this insect, especially with regards to the digestive system. In this study, we analyzed the morphology of the head and buccal apparatus of H. illucens larvae by using optical and scanning electron microscopy, evaluating the different mouthparts and their modifications during larval development. Our analysis showed that the larval head of H. illucens presents similarities to those of campodeiform insect larvae, whereas the mandibular-maxillary complex represents a food intake solution typical of Stratiomyidae that enables BSF larvae to ingest semiliquid food. The mouthparts resemble a "tunnel boring machine", where the hypopharynx separates finer organic particles from coarser and inorganic ones.