Wax Moth Larvae Research Articles

Galleria mellonella as an Antimicrobial Screening Model.

To combat the rising global issue of antibiotic resistance, the accelerated development of novel antibiotics is essential. Current preclinical antimicrobial development yields a significant number of leads that prove unsuitable either prior to or during clinical trials. To increase the efficiency of preclinical development, relevant, standardized, accessible, and cost-effective models must be developed. Galleria mellonella (greater wax moth) larvae are widely used as an infection model to assess microbial virulence, conduct drug toxicity testing, and serve as a preliminary means of evaluating the in vivo efficacy of novel antimicrobial compounds. These infection models have greater biological relevance than many in vitro screens of comparable throughput and decrease reliance on mammalian models when used as a pre-screen for antimicrobial testing. This protocol describes a standardized methodology for the optimization of G. mellonella infection models, which can be applied to bacterial species and antimicrobial therapeutics of choice. Using the WHO priority pathogen Pseudomonas aeruginosa as an exemplar, we outline steps that can be undertaken to develop a reproducible model of infection and therapeutic testing. This includes recommendations on experimental setup, sample preparation, and infection and treatment protocols. Integration of this model within preclinical antimicrobial development pipelines would decrease reliance on mammalian models, reduce the number of ineffective compounds reaching clinical trials, and ultimately increase the efficiency of preclinical antimicrobial development.

Toxicity and Physiological Effect of Lemongrass Essential Oil Nano-capsules on the Greater Wax Moth Larvae Galleria mellonella L. (Lepidoptera: Pyralidae)

Toxicity and Physiological Effect of Lemongrass Essential Oil Nano-capsules on the Greater Wax Moth Larvae Galleria mellonella L. (Lepidoptera: Pyralidae)

Development of Galleria mellonella (wax moth) larvae as a model for snakebite envenoming

Development of Galleria mellonella (wax moth) larvae as a model for snakebite envenoming

The venom of Habrobracon hebetor induces alterations in host metabolism.

The ability of parasitic wasps to manipulate a host's metabolism is under active investigation. Components of venom play a major role in this process. In the present work, we studied the effect of the venom of the ectoparasitic wasp Habrobracon hebetor on the metabolism of the greater wax moth host (Galleria mellonella). We identified and quantified 45 metabolites in the lymph (cell-free hemolymph) of wax moth larvae on the second day after H. hebetor venom injection, using NMR spectroscopy and liquid chromatography coupled with mass spectrometry. These metabolites included 22 amino acids, nine products of lipid metabolism (sugars, amines and alcohols) and four metabolic intermediates related to nitrogenous bases, nucleotides and nucleosides. An analysis of the larvae metabolome suggested that the venom causes suppression of the tricarboxylic acid cycle, an increase in the number of free amino acids in the lymph, an increase in the concentration of trehalose in the lymph simultaneously with a decrease in the amount of glucose, and destructive processes in the fat body tissue. Thus, this parasitoid venom not only immobilizes the prey but also modulates its metabolism, thereby providing optimal conditions for the development of larvae.

The Use of Some Herbal Essential Oils Against Galleria mellonella Larvae and Testing of Bacillus thuringiensis Bacterium Isolated from Galleria mellonella Under Laboratory Conditions

Larvae of wax moths cause great damage in honey bee hives and especially in stored honeycombs. Biological control methods are especially important in the control of wax moth in warehouses, as they do not harm the bee, the product and the environment. This study was carried out to determine the effect of 5%, 10%, 25%, 45% and 55% of peppermint, thyme, nettle seed, and walnut herbal oils and GB1 Bacillus sp. (OR227363) against wax moth larvae (Galleria mellonella) (L1-L3) under laboratory conditions. For each group of fifteen larvae, four herbal oil and one bacterial trials were conducted and two control groups were formed. The trials were conducted in glass jars and the larvae were kept in an oven at 25°C temperature/75% relative humidity. Each jar was checked every day for two weeks and dead/viable larvae were recorded and the dead ones were removed from the jar. As a result of the dose trials, it was determined that the best dose was 2.835x109 cfu/mL for bacteria and 5% concentration of thyme and walnut oil for herbal oil. According to the data obtained, it is thought that GB1 Bacillus sp. isolate can be used as an alternative control method against wax moth larvae.

Speckle tracking phase-contrast computed tomography at an inverse Compton X-ray source.

Speckle-based X-ray imaging (SBI) is a phase-contrast method developed at and for highly coherent X-ray sources, such as synchrotrons, to increase the contrast of weakly absorbing objects. Consequently, it complements the conventional attenuation-based X-ray imaging. Meanwhile, attempts to establish SBI at less coherent laboratory sources have been performed, ranging from liquid metal-jet X-ray sources to microfocus X-ray tubes. However, their lack of coherence results in interference fringes not being resolved. Therefore, algorithms were developed which neglect the interference effects. Here, we demonstrate phase-contrast computed tomography employing SBI in a laboratory-setting with an inverse Compton X-ray source. In this context, we investigate and compare also the performance of the at synchrotron conventionally used phase-retrieval algorithms for SBI, unified modulated pattern analysis (UMPA) with a phase-retrieval method developed for low coherence systems (LCS). We successfully retrieve a full computed tomography in a phantom as well as in biological specimens, such as larvae of the greater wax moth (Galleria mellonella), a model system for studies of pathogens and infections. In this context, we additionally demonstrate quantitative phase-contrast computed tomography using SBI at a low coherent set-up.

Antimicrobial Evaluation of Two Polycyclic Polyprenylated Acylphloroglucinol Compounds: PPAP23 and PPAP53.

Polycyclic polyprenylated acylphloroglucinols (PPAPs) comprise a large group of compounds of mostly plant origin. The best-known compound is hyperforin from St. John's wort with its antidepressant, antitumor and antimicrobial properties. The chemical synthesis of PPAP variants allows the generation of compounds with improved activity and compatibility. Here, we studied the antimicrobial activity of two synthetic PPAP-derivatives, the water-insoluble PPAP23 and the water-soluble sodium salt PPAP53. In vitro, both compounds exhibited good activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium. Both compounds had no adverse effects on Galleria mellonella wax moth larvae. However, they were unable to protect the larvae from infection with S. aureus because components of the larval coelom neutralized the antimicrobial activity; a similar effect was also seen with serum albumin. In silico docking studies with PPAP53 revealed that it binds to the F1 pocket of human serum albumin with a binding energy of -7.5 kcal/mol. In an infection model of septic arthritis, PPAP23 decreased the formation of abscesses and S. aureus load in kidneys; in a mouse skin abscess model, topical treatment with PPAP53 reduced S. aureus counts. Both PPAPs were active against anaerobic Gram-positive gut bacteria such as neurotransmitter-producing Clostridium, Enterococcus or Ruminococcus species. Based on these results, we foresee possible applications in the decolonization of pathogens.

The individual contributions of blaB, blaGOB and blaCME on MICs of β-lactams in Elizabethkingia anophelis.

The blaB, blaGOB and blaCME genes are thought to confer β-lactam resistance to Elizabethkingia anophelis, based on experiments conducted primarily on Escherichia coli. To determine the individual contributions of β-lactamase genes to increased MICs in E. anophelis and to assess their impact on the in vivo efficacy of carbapenem therapy. Scarless gene deletion of one or more β-lactamase gene(s) was performed in three clinical E. anophelis isolates. MICs were determined by broth microdilution. Hydrolytic activity and expressions of β-lactamase genes were measured by an enzymatic assay and quantitative RT-PCR, respectively. In vivo efficacy was determined using Galleria mellonella and murine thigh infection models. The presence of blaB resulted in >16-fold increases, while blaGOB caused 4-16-fold increases of carbapenem MICs. Hydrolysis of carbapenems was highest in lysates of blaB-positive strains, possibly due to the constitutionally higher expression of blaB. Imipenem was ineffective against blaB-positive isolates in vivo in terms of improvement of the survival of wax moth larvae and reduction of murine bacterial load. The deletion of blaB restored the efficacy of imipenem. The blaB gene was also responsible for a >4-fold increase of ampicillin/sulbactam and piperacillin/tazobactam MICs. The presence of blaCME, but not blaB or blaGOB, increased the MICs of ceftazidime and cefepime by 8-16- and 4-8-fold, respectively. The constitutionally and highly expressed blaB gene in E. anophelis was responsible for increased MICs of carbapenems and led to their poor in vivo efficacy. blaCME increased the MICs of ceftazidime and cefepime.

Transplanting human infant gut microbiome species into Galleria mellonella

ObjectiveStudy of the human infant gut microbiome requires the use of surrogate mammalian species such as mice. We sought to investigate the usefulness of the greater wax moth larva, Galleria mellonella, as an alternative.ResultsWe have analysed the native gut microbiome of Galleria and developed methods for clearing the native microbiome and introducing species from human infant faecal samples. We find that some species, e.g. enterococci, are more successful at recolonisation, but that others, e.g. Bifidobacterium, are less so. The work paves the way for using Galleria rather than mice in this and similar work.

Optimization of clonal micropropagation of stone fruit crops

The aim of the research was to optimize the clonal micropropagation of stone crops through the use of improved techniques. The objects of the research at the stage of introduction into culture in vitro were meristematic apexes, at the stage of micro-propagation itself – micro-shoots, at the rooting stage – rooted micro-shoots, at the adaptation stage – micro-plants. All experiments were carried out according to generally accepted methods using the example of the steppe cherry variety Shchedraya, the domestic plum variety Kazanskaya and the hybrid sweet cherry Fatezh. As a result of the research, it was found that the use of a 10 % chlorhexidine solution for sterilization of the initial plant material of stone crops increased the survival rate of apexes on average by 2.4 %; the combined use of growth regulators 6-benzylaminopurine 0.5 mg/l, gibberellic acid 0.2 mg/l, indo-lil-3-butyric acid (IBA) 0.2 mg/l in a nutrient medium and an experimental diode pulsed phytoirradiator in the last proliferation passage activated the proliferation of micro-shoots, increasing the reproduction coefficient of stone crops in 6 passages by an average of 0.9 pcs/explant; the addition of 1.0 mg/l (IBA ) to the nutrient medium provided an increase in the rooting capacity of micro-shoots of stone crops by an average of 9.6 %. The treatment of micro-plants of stone crops at the stage of adaptation by spraying with an 8 % solution of the extract of the products of the larvae of the large wax moth and the use of an experimental LED pulsed phytoirradiator with a mixed spectrum contributed to an increase in their survival rate by an average of 12.4 %. Compliance with the proposed improved methods of clonal micropropagation of stone crops using growth regulators and an experimental LED pulse phytoirradiator with a mixed spectrum made it possible to increase the yield of standard adapted micro-plants by 2 times. At the same time, the cost of one adapted micro-plant decreased by an average of 11.4 rubles, and the profitability of obtaining improved stone crop material increased by 33.7 % and amounted to 160.7 %.

The 3′ UTR of vigR is required for virulence in Staphylococcus aureus and has expanded through STAR sequence repeat insertions

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are alarmingly common, and treatment is confined to last-line antibiotics. Vancomycin is the treatment of choice for MRSA bacteremia, and treatment failure is often associated with vancomycin-intermediate S.aureus isolates. The regulatory 3' UTR of the vigR mRNA contributes to vancomycin tolerance and upregulates the autolysin IsaA. Using MS2-affinity purification coupled with RNA sequencing, we find that the vigR 3' UTR also regulates dapE, a succinyl-diaminopimelate desuccinylase required for lysine and peptidoglycan synthesis, suggesting a broader role in controlling cell wall metabolism and vancomycin tolerance. Deletion of the 3' UTR increased virulence, while the isaA mutant is completely attenuated in a wax moth larvae model. Sequence and structural analyses of vigR indicated that the 3' UTR has expanded through the acquisition of Staphylococcus aureus repeat insertions that contribute sequence for the isaA interaction seed and may functionalize the 3' UTR.

Effect of the Neonicotinoid Insecticide Thiacloprid on Oxidative Stress, Genotoxic, and Immunotoxic Biomarkers in Greater Wax Moth, Galleria mellonella

The aim of this study was to investigate the effect of the neonicotinoid insecticide Thiacloprid on oxidative stress, genotoxic, and immunotoxic biomarkers in Galleria mellonella. The effects of neonicotinoid insecticide thiacloprid on antioxidant enzyme activities, malondialdehyde (MDA) levels, hemocyte number, micronucleus frequency of greater wax moth (Galleria mellonella) larvae at different doses (5, 10, 15, 20, 25, and 30 µg) and periods (24, 48, 72, and 96 hrs) were explored. Superoxide dismutase (SOD) activity increased significantly at 5, 10, and 15 µg thiacloprid doses compared to the control and negative control in all periods tested, while significantly decreased at 20, 25, and 30 µg doses. Catalase (CAT) activity showed significant increases at 5, 10 and 15 µg thiacloprid doses at 24 and 96h compared to the control and negative control. MDA concentrations showed significant increases in all periods compared to the control and negative control. At 24th, 48th, 72nd and 96th, total hemocyte count (THC) decreased significantly at all doses except 5 µg thiacloprid concentration. During all the tested periods, there was a significant increase in the number of micronuclei, particularly at high doses of thiacloprid (20, 25, and 30 µg) compared to both the control and negative control. Additionally, a positive correlation was observed between MDA and the number of micronuclei, while other markers showed a negative correlation with micronucleus (MN). These results suggest that high doses of thiacloprid induce significant increases in micronuclei formation and are positively correlated with MDA levels, indicating oxidative damage and genotoxicity caused by thiacloprid exposure in the tested organism. Overall, our findings suggest that the measured parameters can be considered reliable biomarkers to demonstrate oxidative damage from thiacloprid exposure.

Coexistence of virulent and multidrug-resistant plasmids in an uropathogenic Escherichia coli

ObjectivesThe emergence of pathogens co-harbouring multiple mobile resistance and virulence elements is of great concern in clinical settings. Herein, we report an O101: H10-ST167 Escherichia coli Hu106 strain isolated from the urinary tract of a female in China. MethodsAntibiotic susceptibility testing was used to present the antimicrobial resistance spectrum. Whole-genome sequencing (WGS) and bioinformatic analysis were used to clarify the virulent and resistance mechanisms. Furthermore, the virulence of this strain was tested by the Greater wax moth larvae and siderophore production experiment. ResultsThe strain E. coli Hu106 was resistant to almost all antimicrobials tested, and only susceptible to aztreonam, amikacin, and tigecycline. WGS analysis revealed that the strain Hu106 co-harboured blaNDM-9 and mcr-1 on p2-Hu106, belonging to IncHI2/IncHI2A (256,000 bp). The co-existence of both resistance genes, blaNDM-9 and mcr-1, on the plasmid p2-Hu106 was mainly acquired by transposition recombination of mobile antibiotic elements mediated by IS26 and/or IS1 on IncHI2/IncHI2A type plasmid. In addition, the virulence clusters aerobactin (iutA-iucABCD) and salmochelin (iroBCDEN) were identified on an IncFIB/IncFIC(IncFII) type plasmid p1-Hu106, flanked by small mobile elements such as IS1A, ISkpn28, and IS3, respectively. After performing genomic comparison of p1-Hu106 with the WGS in NCBI, we identified that the virulent plasmid p1-Hu106-like could spread in different clones of E. coli and Klebsiella pneumoniae, revealing its underlying dissemination mechanism between Enterobacterales. Furthermore, the strain caused a decreased survival rate of larvae and produced high siderophore units (62.33%), similar to hypervirulent K. pneumoniae NTUH-K2044. ConclusionsThe strains co-carrying the multidrug-resistant plasmid p2-Hu106 and virulent plasmid p1-Hu106 should be closely monitored to prevent its further spreading.

Characterising phagocytes and measuring phagocytosis from live Galleria mellonella larvae

ABSTRACT Over the last 20 years, the larva of the greater waxmoth, Galleria mellonella, has rapidly increased in popularity as an in vivo mammalian replacement model organism for the study of human pathogens. Experimental readouts of response to infection are most often limited to observing the melanization cascade and quantifying larval death and, whilst transcriptomic and proteomic approaches, and methods to determine microbial load are also used, a more comprehensive toolkit of profiling infection over time could transform the applicability of this model. As an invertebrate, Galleria harbour an innate immune system comprised of both humoral components and a repertoire of innate immune cells – termed haemocytes. Although information on subtypes of haemocytes exists, there are conflicting reports on their exact number and function. Flow cytometry has previously been used to assay Galleria haemocytes, but protocols include both centrifugation and fixation – physical methods which have the potential to affect haemocyte morphology prior to analysis. Here, we present a method for live haemocyte analysis by flow cytometry, revealing that Galleria haemocytes constitute only a single resolvable population, based on relative size or internal complexity. Using fluorescent zymosan particles, we extend our method to show that up to 80% of the Galleria haemocyte population display phagocytic capability. Finally, we demonstrate that the developed assay reliably replicates in vitro data, showing that cell wall β-1,3-glucan masking by Candida albicans subverts phagocytic responses. As such, our method provides a new tool with which to rapidly assess phagocytosis and understand live infection dynamics in Galleria.

Structure-Activity Relationship Study to Develop Peptide Amphiphiles as Species-Specific Antimicrobials.

Antimicrobial peptide amphiphiles (PAs) are a promising class of molecules that can disrupt the bacterial membrane or act as drug nanocarriers. In this study, we prepared 33 PAs to establish supramolecular structure-activity relationships. We studied the morphology and activity of the nanostructures against different Gram-positive and Gram-negative bacterial strains (such as Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Acinetobacter baumannii). Next, we used principal component analysis (PCA) to determine the key contributors to activity. We found that for S. aureus, the zeta potential was the major contributor to the activity while Gram-negative bacteria were more influenced by the partition coefficient (LogP) with the following order P. aeruginosa>E. coli>A. baumannii. We also performed a study of the mechanism of action of selected PAs on the bacterial membrane assessing the membrane permeability and depolarization, changes in zeta potential and overall integrity. We studied the toxicity of the nanostructures against mammalian cells. Finally, we performed an in vivo study using the wax moth larvae to determine the therapeutic efficacy of the active PAs. This study shows cationic PA nanostructures can be an intriguing platform for the development of nanoantibacterials.

Occurrence and Pathogenicity of Indigenous Entomopathogenic Fungi Isolates to Fall Armyworm (Spodoptera frugiperda J. E. Smith) in Western Amhara, Ethiopia

The fall armyworm (FAW) (Spodoptera frugiperda J. E. Smith) is widely recognized as an invasive lepidopteran pest in Africa. Adoption of synthetic pesticides remains an option for emergency control of the FAW, but their large-scale use by small-scale farmers is costly and environmentally unsafe. Management options based on entomopathogenic fungi could minimize risks for health and the environment. In our study, the occurrence of entomopathogenic fungi in different habitats along with their pathogenicity to fall armyworm was examined. A total of 56 soil samples were collected from three locations. A standard isolation method, baited with the wax moth larvae (Galleria mellonella), was used for the isolation of entomopathogenic fungi from soil. Twelve entomopathogenic fungi were isolated from the total soil samples collected. It was observed that Metarhizium spp. was more frequent and widespread than Beauveria spp. and their occurrence was high in the forest habitat. Pathogenicity of indigenous entomopathogenic fungi isolates on fall armyworm larvae was tested by applying conidial suspension containing 1 × 108 spores/ml. All of the isolates tested were pathogenic to larvae of fall armyworm, with mortalities varying from 30 to 80% at 10 days posttreatment. Our results suggest that native entomopathogenic fungi could be integrated for the management of Spodoptera frugiperda larvae.

STUDY OF THE EFFICIENCY AND COMPARATIVE ANALYSIS OF THE APPLICATION OF TECHNIQUES FOR EVALUATING THE QUALITY OF PHYTOPHAGES AND ENTOMOPHAGES AT DIFFERENT STAGES OF ONTOGENESIS

The concept of quality control of insect crops is proposed, which is based on a targeted approach based on the goal and objectives of the implementation of a certain breeding program and is divided into two main directions: control of phytophage production, control of entomophage production. Quality control methods are based on the general biological rule of dependence between the intensity of insect taxis and the overall viability of biomaterial. The effectiveness of methods for assessing the quality of phytophages (on the example of the mother cultures of the grain moth Sitotroga cerealella, the wax moth Galleria mellonella) and entomophages (laboratory and natural populations of P. bioculatus.) of the Center for Mother Cultures of Insects Engineering and Technological Institute “Biotekhnica” of the National Academy of Sciences (ETI “Biotekhnica”) at different stages of ontogenesis was investigated using the indicator the intensity of taxis. As a result of research conducted on the culture of phytophagous, grain moth, a significant difference with the control was established in the indicators of viability of eggs, larvae, pupae and adults: 25; 6; 33 and 11%, respectively. Studies conducted with another phytophagous representative – the wax firefly – also showed positive results (16; 17; 16; 11%, respectively). The obtained (as a result of preliminary quality control) and selection culture had significantly higher viability indicators at all stages of ontogenesis. It was established that the intensity of chemotaxis of larvae of the 1st age of entomophages directly correlates with the indicator of viability of the insect culture. When selecting the starting material for successful culture, it is possible to select by chemotaxis of the larvae after rebirth from the egg. Thus, viability when selecting for Colorado beetle eggs significantly (p<0.01) exceeds this indicator in other selection options – Colorado beetle larvae and wax firefly larvae: 82; 59; 73% respectively. When working with an artificial population of P. bioculatus, on wax moth larvae, the highest intensity of taxis was noted in the variant with wax moth larvae compared to Colorado beetle eggs and Colorado beetle larvae – 37; 34; 29% respectively. This indicates a shift in the number of the population in the direction of individuals that successfully feed on unnatural food. Their viability was significantly higher than the viability in other variants of the study with an artificial population – 78; 71; 68% respectively. The proposed method of selection of starting material for cultivation of P. bioculatus allows differentiation of individuals for further effective reproduction, and the use of such biomaterial in programs of biological protection of plants from pests. A comparative analysis of methods for assessing the quality of phytophagous and entomophagous cultures at different stages of ontogenesis was carried out using the indicator of the overall viability of insect populations. It is an integrated indicator that takes into account the viability of the population at all stages of its development. All tested objects of the study showed that control and improvement of culture quality by the proposed method lead to an increase in the viability of biomaterial by 18.71; 18.56 and 8.86%, respectively (egg, larva, pupa).

Analysis of comb-gnawing behavior in Apis cerana cerana (Hymenoptera: Apidae).

Apis cerana cerana exhibits a prominent biological trait known as comb gnawing. In this study, gnawed combs from colonies during different seasons were collected, investigating the comb age and locations of gnawing. Patterns of comb gnawing were recorded, and the effects of 2 factors, namely, comb type and season, on the mass of wax residues and the gnawed surface area were measured. The results revealed that A. c. cerana predominantly gnaws combs that have been used for over 6 months, with gnawing concentrated in the brood-rearing area. In the first 3 seasons, significantly higher masses of wax residues and larger gnawed surface areas were found in greater wax moth larvae (GWML)-infested combs compared to newly built and old combs. Also, there were significantly higher masses and areas gnawed by A. c. cerana in old combs compared to newly built combs in all 4 seasons. Compared to other seasons, it exhibited significantly higher masses and areas resulting from comb-gnawing in newly built or old combs in winter. However, there were no significant differences in the masses of wax residues and surface areas gnawed in GWML-infested combs across the first 3 seasons. In conclusion, this study documented the impact of comb type and season on the comb-gnawing behavior of A. c. cerana, contributing to beekeeping management practices and the current understanding of bee biology.

Hedonic rating coupled with sensory profiles using CATA for six whole roasted or deep-fried insects among Danish 11-13-year-old children

The consumption of insects is not rooted in traditional Western culture, which creates a barrier to integrate insects into our diets, especially among preadolescents. This study was executed during a Danish science outreach week to educate children on eating insects through educational activities. 11–13-y.o. children were invited to taste six different types of whole roasted or deep-fried insects (grasshoppers, cricket, bee larvae, wax moth larvae, buffalo worms and mealworms), asked to rate their liking (15-point hedonic scale) and describe the sensory properties for each insect (50 sensory attributes using Check-All-That-Apply format). Food neophobia was assessed using the Food Neophobia Test Tool to explore the link between food neophobia and hedonic responses of insects. Results showed significant differences in sensory and hedonic responses (p < 0.0001, respectively) between insect types. In general, insects showed low mean liking. They were distributed in two significantly different groups. Mealworms (6.9), buffalo worms (6.6), and bee larvae (6.4) had higher mean liking compared to crickets (5.2), wax moth larvae (5.1), and grasshoppers (4.7). Furthermore, 19 descriptors discriminated between insects demonstrating that insects vary in sensory characteristics. “Crunchy” was significantly associated with liking, highlighting the importance of this sensory attribute when promoting the consumption of edible insects in preadolescents. Insect liking decreased with increasing levels of food neophobia, which was also affected by age- and gender. The study offers new insights into the acceptance and sensory properties of commercial insects that can be used to promote insect consumption and for future product development targeted at this group.

Transcription Factors Tec1 and Tec2 Play Key Roles in the Hyphal Growth and Virulence of Mucor lusitanicus Through Increased Mitochondrial Oxidative Metabolism.

Mucormycosis is a lethal and difficult-to-treat fungal infection caused by fungi of the order Mucorales. Mucor lusitanicus, a member of Mucorales, is commonly used as a model to understand disease pathogenesis. However, transcriptional control of hyphal growth and virulence in Mucorales is poorly understood. This study aimed to investigate the role of Tec proteins, which belong to the TEA/ATTS transcription factor family, in the hyphal development and virulence of M. lusitanicus. Unlike in the genome of Ascomycetes and Basidiomycetes, which have a single Tec homologue, in the genome of Mucorales, two Tec homologues, Tec1 and Tec2, were found, except in that of Phycomyces blakesleeanus, with only one Tec homologue. tec1 and tec2 overexpression in M. lusitanicus increased mycelial growth, mitochondrial content and activity, expression of the rhizoferrin synthetase-encoding gene rfs, and virulence in nematodes and wax moth larvae but decreased cAMP levels and protein kinase A (PKA) activity. Furthermore, tec1- and tec2-overexpressing strains required adequate mitochondrial metabolism to promote the virulent phenotype. The heterotrimeric G beta subunit 1-encoding gene deletant strain (Δgpb1) increased cAMP-PKA activity, downregulation of both tec genes, decreased both virulence and hyphal development, but tec1 and tec2 overexpression restored these defects. Overexpression of allele-mutated variants of Tec1(S332A) and Tec2(S168A) in the putative phosphorylation sites for PKA increased both virulence and hyphal growth of Δgpb1. These findings suggest that Tec homologues promote mycelial development and virulence by enhancing mitochondrial metabolism and rhizoferrin accumulation, providing new information for the rational control of the virulent phenotype of M. lusitanicus.