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In broiler production, numerous feed additives have been utilized to promote growth, feed efficiency, immunity, and antioxidant capacity. Similarly, various techniques, including electric fans, cooling pad systems, and water sprinkling through foggers, have been employed to mitigate heat stress in poultry. However, due to cost constraints, these methods may not always be feasible, and alternative nutritional strategies can be followed, including the use of balancing of nutrients. One such nutritional strategy for reducing stress is the incorporation of Tenebrio molitor (mealworm) as a feed additive. Therefore, this study was designed to investigate the impact of T. molitor as a feed supplement on the performance, immunity, and economics of heat-stressed broilers. A total of 150 (day-old) broilers were randomly allocated to five dietary groups: such as D0 (control), D1, D2, D3, and D4, provided with T. molitor at the rate of 0, 0.5, 1.0, 1.5, and 2 g/kg of feed, respectively. The broilers were exposed to temperatures ranging from 32-38°C during the trial. Results showed the highest significant total weight gain, improved feed conversion ratio, and dressing percentage, and reduced feed intake in the D4 group compared to all other groups. Similarly, the highest antibody titers against the Newcastle disease, and lower mortality rate were calculated in the D4 group than in the remaining groups. Meat quality was not affected. A significant increase in the net profit was observed in the D4 group as compared to the leftover groups. Based on the current findings, it is concluded that T. molitor at the rate of 2.0 g/kg in broiler feed improves the production performance, boosts the immune response against the Newcastle disease, and increases the economic return. [ABSTRACT FROM AUTHOR]

Simple Summary: Research on alternative protein sources is becoming increasingly important in our modern world due to the issue of sustainability and efficiency in food production. Insects are seen as promising alternatives to known animal protein sources due to their attractive nutritional composition and the fact that they can be bred and fattened in a possibly more resource-efficient way. As consumers, especially in Europe, have a certain disgusting feeling towards whole insects (larvae), various research projects are working on the production of hybrid (meat) products in which the insects are mostly processed as dried powders or flour so that they are unrecognizable to the consumer as a whole. In this study, in order to avoid the high-energy and complex steps of pre-processing, such as protein or fat extraction, drying, or hydrolysis, whole larvae of the species Alphitobius (A.) diaperinus (Alphitobius diaperinus, Panzer, 1797) and Tenebrio (T.) molitor (Tenebrio molitor, Linnaeus, 1758) were pulverized and then processed in a meat grinder (without pre-processing) together with the lean meat and other required ingredients. Five batches of two types of cooked sausages (turkey and pork cooked sausages) were produced: in addition to a control sausage without insect content, 10% and 20% of the turkey and pork lean meats were replaced with A. diaperinus and/or T. molitor larvae powder. The replacement with insect larvae had an influence on the color of the sausages, as well as on sensory properties such as smell and appearance. There was no effect on texture and nutritional values (e.g., fat and protein content). The processing of the insect larvae in the meat product also had no effect on the growth of inoculated bacterial species over a storage period. Proteins from insect production represent an interesting (environmentally friendly) option or supplement to commercial livestock farming. At present, however, the larval stages of T. molitor (mealworm) and A. diaperinus (buffalo worm) have been authorized as food for human consumption EU-wide, as have the nymph and adult stages of Locusta (L.) migratoria (Locusta migratoria, Linnaeus, 1758) and Acheta (A.) domesticus (house cricket, Acheta domesticus, Linnaeus, 1758). However, there is the problem that insects that are recognizable as a whole tend to be avoided by consumers, especially in the European region, as they are reminiscent of living things and can cause aversion and disgust in consumers. Against this background, in the present study, five batches of two types of cooked sausages were produced: on the one hand, with turkey, and on the other hand, with pork lean meat as a base. In different formulations, 10% and 20% of the meat contents (turkey or pork) in these meat products were replaced by deep-frozen, pulverized T. molitor and A. diaperinus larvae. The effects of the addition of these insects in the products on the microbiological and physicochemical parameters of these cooked sausages, compared to a product without insect content, directly after heating, were investigated. After production, a storage trial was also carried out to determine whether possible insect ingredients could influence the growth of inoculated bacterial species (Bacillus (B.) cereus, Escherichia (E.) coli, Listeria (L.) monocytogenes, and Campylobacter (C.) jejuni) and how the addition of insect larvae affectsthe sensory and physicochemical properties during storage. The study showed that the products with insects had reduced lightness (turkey p C = 0.025), increased yellowness (pork p S = 0.0009, p C < 0.0001 and turkey p C = 0.0027) and a reduced red color (pork p S < 0.0001, p C = 0.0001) after heating when compared to the cooked sausages without insects. However, no significant differences between the various cooked sausages with or without insects in terms of cooking loss, firmness, and protein, ash, and fat or water contents were found. The microbiological tests showed, on the one hand, that the prior microbial reduction (e.g., in the form of blanching) of the insect larvae was essential in order to guarantee the flawless microbiological quality of the cooked sausages and, on the other hand, that the addition of insects to the cooked sausages did not significantly affect the growth of the inoculated bacterial species and that no sensory differences could be detected during storage. Despite the significant color effects on the product, A. diaperinus and T. molitor larvae would be suitable as protein or meat alternatives in cooked sausages, but they would have to undergo pre-treatment, primarily with regard to microbiological safety. The extent to which a complete replacement of meat is possible has to be investigated in further studies. [ABSTRACT FROM AUTHOR]

Abstract Tenebrio molitor, which is rich in various nutrients, and its polysaccharides, as significant bioactive substances, exhibit strong antioxidant effects. This study utilized defatted T. molitor as raw material and employed an ultrasound-assisted extraction method. The factors considered include extraction temperature, time, ultrasound power, and liquid-to-feed ratio, with the yield of T. molitor polysaccharides as the response value. Based on single-factor experiments and response surface methodology, the optimal extraction parameters for T. molitor polysaccharides were determined. Following purification, protein removal, and dialysis to eliminate impurities, the structure of the extracted polysaccharides was preliminarily investigated using infrared spectroscopy. Their antioxidant activities were explored by measuring their DPPH·, OH·, and ABTS+· radical scavenging abilities and Fe3+ reducing power. The results indicated that the optimal conditions for ultrasound-assisted extraction were an extraction temperature of 75 °C, an extraction time of 150 min, an ultrasound power of 270 W, and a liquid-to-feed ratio of 15:1 mL/g, yielding a polysaccharide extraction rate of 9.513%. Infrared spectroscopy analysis revealed the presence of pyranose sugars with main functional groups including C-O, C=O, and O-H. Antioxidant activity tests showed that within a certain concentration range, the higher the polysaccharide concentration, the stronger its radical scavenging abilities. Compared with Vitamin C(Vc), the polysaccharides had stronger scavenging abilities for DPPH· and OH·, some scavenging ability for ABTS+·, and Fe3+ reduction ability, and corresponding to IC50 values of 0.9625, 9.1909, and 235.69 mg/mL respectively. The Fe3+ reducing power reached a maximum absorbance of 0.38899 at a concentration of 1.6 mg/mL. T. molitor polysaccharides demonstrate promising antioxidant activity and potential as functional ingredients in food, health products, and pharmaceuticals, providing new technical references for the development and utilization of T. molitor resources.

The objective of this study was to explore the impacts of various ultrasonic powers (0, 300, 500, 700, and 900 W) on the structure and functional attributes of the myofibrillar protein (MP) of Tenebrio molitor. As the ultrasonic intensity escalated, the extraction efficiency and yield of the MP rose, while the particle size and turbidity decreased correspondingly. The reduction in sulfhydryl group content and the increase in carbonyl group content both suggested that ultrasonic treatment promoted the oxidation of the MP to a certain extent, which was conducive to the formation of a denser and more stable gel network structure. This was also affirmed by SEM images. Additionally, the findings of intrinsic fluorescence and FTIR indicated that high-intensity ultrasound significantly altered the secondary structure of the protein. The unfolding of the MP exposed more amino acid residues, the α-helix decreased, and the β-helix improved, thereby resulting in a looser and more flexible conformation. Along with the structural alteration, the surface hydrophobicity and emulsification properties were also significantly enhanced. Besides that, SDS–PAGE demonstrated that the MP of T. molitor was primarily composed of myosin heavy chain (MHC), actin, myosin light chain (MLC), paramyosin, and tropomyosin. The aforementioned results confirmed that ultrasonic treatment could, to a certain extent, enhance the structure and function of mealworm MP, thereby providing a theoretical reference for the utilization of edible insect proteins in the future, deep-processing proteins produced by T. molitor, and the development of new technologies. [ABSTRACT FROM AUTHOR]

The objective of this study was to explore the impacts of various ultrasonic powers (0, 300, 500, 700, and 900 W) on the structure and functional attributes of the myofibrillar protein (MP) of Tenebrio molitor. As the ultrasonic intensity escalated, the extraction efficiency and yield of the MP rose, while the particle size and turbidity decreased correspondingly. The reduction in sulfhydryl group content and the increase in carbonyl group content both suggested that ultrasonic treatment promoted the oxidation of the MP to a certain extent, which was conducive to the formation of a denser and more stable gel network structure. This was also affirmed by SEM images. Additionally, the findings of intrinsic fluorescence and FTIR indicated that high-intensity ultrasound significantly altered the secondary structure of the protein. The unfolding of the MP exposed more amino acid residues, the α-helix decreased, and the β-helix improved, thereby resulting in a looser and more flexible conformation. Along with the structural alteration, the surface hydrophobicity and emulsification properties were also significantly enhanced. Besides that, SDS–PAGE demonstrated that the MP of T. molitor was primarily composed of myosin heavy chain (MHC), actin, myosin light chain (MLC), paramyosin, and tropomyosin. The aforementioned results confirmed that ultrasonic treatment could, to a certain extent, enhance the structure and function of mealworm MP, thereby providing a theoretical reference for the utilization of edible insect proteins in the future, deep-processing proteins produced by T. molitor, and the development of new technologies.

In recent years, insect meal has attracted increasing interest as an innovative protein source to replace fish meal in feed formulations due to its valuable nutritional profile. This research aimed to compare the effects of different levels of dietary inclusion of the yellow mealworm beetle (T. molitor) larvae meal on the sensory quality of rainbow trout (Oncorhynchus mykiss) fillets and retrospectively on the acceptability of this protein source to consumers. The results showed that the inclusion of T. molitor larvae meal did not induce sensory changes in the trout fillets, while regarding consumer acceptability and willingness to buy and pay, it was shown that a certain level of rejection towards this alternative protein still exists. The work described in this scientific manuscript adds more knowledge on the study of consumer acceptability of this protein source. [ABSTRACT FROM AUTHOR]

In the study of fungal pathogenesis, alternative methods have gained prominence due to recent global legislation restricting the use of mammalian animals in research. The principle of the 3 Rs (replacement, reduction, and refinement) is integrated into regulations and guidelines governing animal experimentation in nearly all countries. This principle advocates substituting vertebrate animals with other invertebrate organisms, embryos, microorganisms, or cell cultures. This review addresses host–fungus interactions by employing three-dimensional (3D) cultures, which offer more faithful replication of the in vivo environment, and by utilizing alternative animal models to replace traditional mammals. Among these alternative models, species like Caenorhabditis elegans and Danio rerio share approximately 75% of their genes with humans. Furthermore, models such as Galleria mellonella and Tenebrio molitor demonstrate similarities in their innate immune systems as well as anatomical and physiological barriers, resembling those found in mammalian organisms. [ABSTRACT FROM AUTHOR]

Larva dari Tenebrio molitor belum dimanfaatkan secara maksimal, sementara secara alami memiliki manfaat sebagai pengurai senyawa organik dan anorganik di ekosistem. Penelitian ini bertujuan untuk menghitung rerata pertambahan panjang dan bobot, laju degradasi,dan nilai indeks reduksi (WRI) beberapa jernis paper pulp menggunakan larva T. molitor. Penelitian menggunakan Rancangan Acak Lengkap (RAL) dengan 10 larva T. molitor instar 8. Pakan yang diberikan terdiri dari tisu makan, tisu basah, dan puntung rokok. Larva kontrol diberi pakan dedak. Masing-masing pakan diberikan sebanyak 1 gram dan diulang lima kali. Larva diukur pertambahan panjang dan beratnya 3 hari sekali. Pergantian pakan dilakukan dalam 3 hari, apabila sebelum 3 hari pakan yang diberikan habis, maka larva diberi pakan sebanyak 1 gram. Analisis data yang digunakan pada penelitian ini adalah analisis inferensial dengan taraf kepercayaan 5%. Nilai tertinggi pertambahan panjang didapatkan pada pakan tisu basah dan putung rokok sebesar 1.58 gr sedangkan terendah pakan tisu makan sebesar 1.41 gr. Nilai tertinggi pertambahan bobot larva pada pakan tisu basah (0.06 gr), sedangkan terendah pakan tisu makan dan putung rokok (0.05 gr). Nilai tertinggi laju degradasi pada pakan tisu makan (0.88), sedangkan terendah pada pakan tisu basah (0.45). Nilai tertinggi WRI pada pakan putung rokok (0.49), sedangakn terendah pada pakan tisu makan (0.23). Kesimpulan dari penelitian ini bahwasanya penggunaan larva T. molitor bisa mendegradasi berbagai olahan paper pulp.

In recent years, insect meal has attracted increasing interest as an innovative protein source to replace fish meal in feed formulations due to its valuable nutritional profile. This research aimed to compare the effects of different levels of dietary inclusion of the yellow mealworm beetle (T. molitor) larvae meal on the sensory quality of rainbow trout (Oncorhynchus mykiss) fillets and retrospectively on the acceptability of this protein source to consumers. The results showed that the inclusion of T. molitor larvae meal did not induce sensory changes in the trout fillets, while regarding consumer acceptability and willingness to buy and pay, it was shown that a certain level of rejection towards this alternative protein still exists. The work described in this scientific manuscript adds more knowledge on the study of consumer acceptability of this protein source.

Previous studies have shown that larvae of the yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae), need a source of moisture to grow and perform well. Currently, much research has been oriented towards the effect of dry feed on larval growth and performance. The effect of different wet feeds as moisture source on the performance traits of T. molitor larvae has not been thoroughly investigated yet. This study aims to investigate in laboratory trials the effect of various gelling agents (agar, carrageenans, guar gum, xanthan gum, sodium alginate, modified starch, and pectin) on the growth and performance of T. molitor larvae. A number of 50 newly emerged larvae obtained from the rearings of the LEAZ were inserted in plastic vials together with 4 g of wheat bran as dry feed. Additionally, 1 g of gelling agents was provided 3 times per week as moisture sources. Carrot slices served as control. Larval survival and weight were recorded weekly until the appearance of the first pupa. Dry feed was replenished when depleted. Our data showed that gelling agents efficiently supported the growth of T. molitor larvae, in terms of larval survival and weight, as well as feed utilization expressed as FCR. Interestingly, carrageenans seem to be the most appropriate gelling agent for T. molitor larvae rearing as it can enhance their weight and is also able to reduce their development time and their specific growth rate. [Display omitted] • Gelling agents were used as a moisture source for the rearing of T. molitor larvae. • Gelling agents were a suitable moisture source that did not impact larvae growth. • Carrot slices were the most suitable moisture source. • Carrageenan reduced development time of the larvae. [ABSTRACT FROM AUTHOR]

Abstract In the case of Tenebrionidae family insects, studies focus on larval stage, leaving a lack of information regarding other stages. Therefore, this study was performed in order to understand the differences between the nutritional composition and the bioactivity of two species of this family in their adult stage, fed with a specific diet. Adult beetles of both species were defatted, lyophilized and protein extracted with buffer. Proximal and phytochemical analysis of the extracts of each insect were performed, along with protein extract and hydrolysis analysis by Tris-Tricine and Tris Glycine SDS PAGE. This analysis showed that T. molitor contained more protein and fat than U. dermestoides but contained less crude fiber. The protein extraction was made with PBS, where 130 and 45 kDa bands showed predominant for U. dermestoides, and less protein was present for T. molitor. Antioxidant and antimicrobial activities of the enzymatic protein hydrolysates and protein crude extracts were determined. Presence of protein associated with the antioxidant activity were found in both insects. Nonetheless U. dermestoides had a higher antioxidant activity with the protein extract in contrast with the higher antioxidant activity shown by U. dermestoides once the extracts were digested. After proteolysis, protein extracts showed an increasing antioxidant activity, plus, the ability to inhibit microbial growth of Proteus, Shigella and Bacillus. Insect protein hydrolysates with protease open the possibility for the use of these beetles as new sources of encrypted peptides for microbiological control once characterized.

Innate immunity is the ultimate line of defense against invading pathogens in insects. Unlike in the mammalian model, in the insect model, invading pathogens are recognized by extracellular receptors, which activate the Toll signaling pathway through an extracellular serine protease cascade. In the Toll-NF-κB pathway, the extracellular spätzle protein acts as a downstream ligand for Toll receptors in insects. In this study, we identified a novel Spätzle isoform (Tm Spz1b) from RNA sequencing database of Tenebrio molitor. Tm Spz1b was bioinformatically analyzed, and functionally characterized for the antimicrobial function by RNA interference (RNAi). The 702 bp open reading frame of Tm Spz1b encoded a putative protein of 233 amino acid residues. A conserved cystine-knot domain with seven cysteine residues in Tm Spz1b was involved in three disulfide bridges and the formation of a spätzle dimer. TmSpz1b was mostly expressed in the hemocytes of T. molitor late instar larvae. The mRNA expression of TmSpz1b was highly induced in the hemocytes after Escherichia coli , Staphylococcus aureus , and Candida albicans stimulation of T. molitor larvae. TmSpz1b silenced larvae were significantly more susceptible to E. coli infection. In addition, RNAi-based functional assay characterized Tm Spz1b to be involved in the positive regulation of antimicrobial peptide genes in hemocytes and fat bodies. Further, the TmDorX2 transcripts were downregulated in TmSpz1b silenced individuals upon E. coli challenge suggesting the relationship to Toll signaling pathway. These results indicate that Tm Spz1b is involved in the T. molitor innate immunity, causes the sequestration of Gram-negative bacteria by the regulatory action of antimicrobial peptides, and enhances the survival of T. molitor larvae. [ABSTRACT FROM AUTHOR]

Innate immunity is the ultimate line of defense against invading pathogens in insects. Unlike in the mammalian model, in the insect model, invading pathogens are recognized by extracellular receptors, which activate the Toll signaling pathway through an extracellular serine protease cascade. In the Toll-NF-κB pathway, the extracellular spätzle protein acts as a downstream ligand for Toll receptors in insects. In this study, we identified a novel Spätzle isoform (TmSpz1b) from RNA sequencing database of Tenebrio molitor. TmSpz1b was bioinformatically analyzed, and functionally characterized for the antimicrobial function by RNA interference (RNAi). The 702 bp open reading frame of TmSpz1b encoded a putative protein of 233 amino acid residues. A conserved cystine-knot domain with seven cysteine residues in TmSpz1b was involved in three disulfide bridges and the formation of a spätzle dimer. TmSpz1b was mostly expressed in the hemocytes of T. molitor late instar larvae. The mRNA expression of TmSpz1b was highly induced in the hemocytes after Escherichia coli, Staphylococcus aureus, and Candida albicans stimulation of T. molitor larvae. TmSpz1b silenced larvae were significantly more susceptible to E. coli infection. In addition, RNAi-based functional assay characterized TmSpz1b to be involved in the positive regulation of antimicrobial peptide genes in hemocytes and fat bodies. Further, the TmDorX2 transcripts were downregulated in TmSpz1b silenced individuals upon E. coli challenge suggesting the relationship to Toll signaling pathway. These results indicate that TmSpz1b is involved in the T. molitor innate immunity, causes the sequestration of Gram-negative bacteria by the regulatory action of antimicrobial peptides, and enhances the survival of T. molitor larvae.

The industrial farming of edible insects for food and feed generally employs cultivated plants and derivatives as rearing diets. These feed substrates may contain toxins produced by different species of fungi which cause adverse health effects to consumers. Frequently found in cereals and cereal products, mycotoxins aflatoxins, ochratoxins and fumonisins are harmful to human and animal health. In this study the uptake of aflatoxin B1 (AFB1), ochratoxin A (OTA) and fumonisin B1 (FB1) and their elimination rates were investigated in T. molitor larvae fed on cereal-based diets contaminated with two concentration levels for each mycotoxin. New analytical methods for extraction and quantification of AFB1, OTA and FB1 residues in larvae were developed and validated. It has been demonstrated that T. molitor larvae did not accumulate in detectable or dangerous concentrations the three tested mycotoxins at the two tested concentration levels and that a fasting period of 24 hours ensured a sufficient elimination rate of AFB1, OTA and FB1 residues. These results represent a preliminary investigation to define an appropriate quality control procedure to assess the safe use of T. molitor for food and feed purposes. [ABSTRACT FROM AUTHOR]

The cystine knot protein Spätzle is a Toll receptor ligand that modulates the intracellular signaling cascade involved in the nuclear factor kappa B (NF-κB)-mediated regulation of antimicrobial peptide (AMP)-encoding genes. Spätzle-mediated activation of the Toll pathway is critical for the innate immune responses of insects against Gram-positive bacteria and fungi. In this study, the open reading frame (ORF) sequence of Spätzle-like from T. molitor (TmSpz-like) identified from the RNA sequencing dataset was cloned and sequenced. The 885-bp TmSpz-like ORF encoded a polypeptide of 294 amino acid residues. TmSpz-like comprised a cystine knot domain with six conserved cysteine residues that formed three disulfide bonds. Additionally, TmSpz-like exhibited the highest amino acid sequence similarity with T. castaneum Spätzle (TcSpz). In the phylogenetic tree, TmSpz-like and TcSpz were located within a single cluster. The expression of TmSpz-like was upregulated in the Malpighian tubules and gut tissues of T. molitor. Additionally, the expression of TmSpz-like in the whole body and gut of the larvae was upregulated at 24 h post-E. coli infection. The results of RNA interference experiments revealed that TmSpz-like is critical for the viability of E. coli-infected T. molitor larvae. Eleven AMP-encoding genes were downregulated in the E. coli-infected TmSpz-like knockdown larvae, which suggested that TmSpz-like positively regulated these genes. Additionally, the NF-κB-encoding genes (TmDorX1, TmDorX2, and TmRelish) were downregulated in the E. coli-infected TmSpz-like knockdown larvae. Thus, TmSpz-like plays a critical role in the regulation of AMP production in T. molitor in response to E. coli infection. [ABSTRACT FROM AUTHOR]

In the case of Tenebrionidae family insects, studies focus on larval stage, leaving a lack of information regarding other stages. Therefore, this study was performed in order to understand the differences between the nutritional composition and the bioactivity of two species of this family in their adult stage, fed with a specific diet. Adult beetles of both species were defatted, lyophilized and protein extracted with buffer. Proximal and phytochemical analysis of the extracts of each insect were performed, along with protein extract and hydrolysis analysis by Tris-Tricine and Tris Glycine SDS PAGE. This analysis showed that T. molitor contained more protein and fat than U. dermestoides but contained less crude fiber. The protein extraction was made with PBS, where 130 and 45 kDa bands showed predominant for U. dermestoides, and less protein was present for T. molitor. Antioxidant and antimicrobial activities of the enzymatic protein hydrolysates and protein crude extracts were determined. Presence of protein associated with the antioxidant activity were found in both insects. Nonetheless U. dermestoides had a higher antioxidant activity with the protein extract in contrast with the higher antioxidant activity shown by U. dermestoides once the extracts were digested. After proteolysis, protein extracts showed an increasing antioxidant activity, plus, the ability to inhibit microbial growth of Proteus, Shigella and Bacillus. Insect protein hydrolysates with protease open the possibility for the use of these beetles as new sources of encrypted peptides for microbiological control once characterized. [ABSTRACT FROM AUTHOR]