Principle Investigation and Method Standardization of Inhibition Zone Assay Based on Antimicrobial Peptides Extracted from Black Soldier Fly Larvae
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ABSTRACT:
SUBMITTER: Shen W
PROVIDER: S-EPMC11348260 | biostudies-literature | 2024 Aug
REPOSITORIES: biostudies-literature
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Project description:Antibacterial peptides from black soldier fly larvae extract were prepared using Flash column chromatography. Three out of five fractions (F2, F3 and F4) showed antibacterial activity against Listeria monocytogenes DMST 17303 with a minimum inhibitory concentration (MIC) of 1 mM, followed by Salmonella enterica Enteritidis DMST 15679 and Escherichia coli O157:H7 DMST 12743 with a MIC ranging from 4 to 8 mM. Due to the higher yield, F2 and F3 were further analyzed on their mode of action against L. monocytogenes DMST 17303. Both fractions, particularly F2, exerted antibacterial activity through inducing bacterial cell membrane disintegration and interaction with intracellular compounds including fatty acids, proteins, and nucleic acids. The F3 did not show significant hemolytic activity up to 4 mM, while F2 showed lower than 5% hemolysis up to 8 mM. Time-to-kill analysis revealed that F2 was more effective and exerted a sustainable killing effect after 2 and 4 h, depending on the concentration of 1 and 2×MIC, respectively, while the F3 at 2×MIC could completely kill the test bacteria within 24 h. Among the identified peptides in the fractions, those with charged, either positively or negatively, and moderate hydrophobicity of ranging 6.68-15.70, namely CGPPRQGPFPR, HLEEELK, LEEAEERAD, TEELEEAKKK, and KGNSELEEAKKK, are potential antimicrobial peptides.
| S-EPMC11531597 | biostudies-literature
Project description:BackgroundProtein hydrolysates from insects are recognized for their biological activities. Black soldier fly larvae (BSFL) have drawn attention due to their antioxidant protein hydrolysates. However, research on bioactive peptides derived from these hydrolysates, particularly their cancer chemopreventive potential, remains limited. This study aims to evaluate the antioxidant, anti-inflammatory, antimutagenic, and anticancer activities of BSFL-derived bioactive peptides and explore the molecular mechanisms.MethodsAlkali-soluble BSFL protein (ASBP) was extracted and hydrolyzed using Alcalase and bromelain under optimized conditions. Antioxidant activity was assessed via FRAP, ABTS, and DPPH assays. The hydrolysate with the highest antioxidant activity was fractionated into molecular weight (MW) groups (>30, 10, and <3 kDa). The bioactivity of fractionated peptides was evaluated through antioxidant, anti-inflammatory (nitric oxide production in RAW 264.7 cells), antimutagenic (Ames test), and anticancer (CCK-8 assay on HCT 116, COLO205, Cw-2, and Caco-2 cells) assays. Mechanistic insights were obtained via microarray and Western blot analyses. Peptides were identified by LC-MS/MS.ResultsThe ASBP-Alcalase hydrolysate (ASBP-AH) showed optimal antioxidant activity at 3% (w/w) for 4 h. The ASBP-AH 30 (MW > 30 kDa) fraction exhibited the highest antioxidant capacity. In contrast, the ASBP-AH3 (MW < 3 kDa) fraction exhibited significant antimutagenic effects, reduced nitric oxide production, and decreased COLO205 cell viability. Treatment with ASBP-AH3 at its LC50 dose modulated the SKP2/p21/cyclin D1 pathways. Mostly peptides from ASBP-AH3 were composed of hydrophobic and charged amino acids.ConclusionsBSFL-derived bioactive peptides exhibit potential as multifunctional agents for cancer chemoprevention. In vivo studies are required to explore their clinical applications.
| S-EPMC11858422 | biostudies-literature
Project description:The black soldier fly is a non-pest insect of interest to the sustainability community due to the high eating rates of its edible larvae. When found on carcases or piles of rotting fruit, this larva often outcompetes other species of scavengers for food. In this combined experimental and theoretical study, we elucidate the mechanism by which groups of black soldier fly larvae can eat so quickly. We use time-lapse videography and particle image velocimetry to investigate feeding by black soldier fly larvae. Individually, larvae eat in 5 min bursts, for 44% of the time, they are near food. This results in their forming roadblocks around the food, reducing the rate that food is consumed. To overcome these limitations, larvae push each other away from the food source, resulting in the formation of a fountain of larvae. Larvae crawl towards the food from below, feed and then are expelled on the top layer. This self-propagating flow pushes away potential roadblocks, thereby increasing eating rate. We present mathematical models for the rate of eating, incorporating flow rates measured from our experiments.
| S-EPMC6408359 | biostudies-literature
Project description:Unlike for vertebrates, the impact of starvation on the gut microbiome of invertebrates is poorly studied. Deciphering shifts in metabolically active associated bacterial communities in vertebrates has led to determining the role of the associated microbiome in the sensation of hunger and discoveries of associated regulatory mechanisms. From an invertebrate perspective, such as the black soldier fly, such information could lead to enhanced processes for optimized biomass production and waste conversion. Bacteria associated with food substrates of black soldier fly are known to impact corresponding larval life-history traits (e.g., larval development); however, whether black soldier fly larval host state (i.e., starved) impacts the gut microbiome is not known. In this study, we measured microbial community structural and functional shifts due to black soldier fly larvae starvation. Data generated demonstrate such a physiological state (i.e., starvation) does in fact impact both aspects of the microbiome. At the phylum level, community diversity decreased significantly during black soldier fly larval starvation (p = 0.0025). Genus level DESeq2 analysis identified five genera with significantly different relative abundance (q < 0.05) across the 24 and 48 H post initiation of starvation: Actinomyces, Microbacterium, Enterococcus, Sphingobacterium, and Leucobacter. Finally, we inferred potential gene function and significantly predicted functional KEGG Orthology (KO) abundance. We demonstrated the metabolically active microbial community structure and function could be influenced by host-feeding status. Such perturbations, even when short in duration (e.g., 24 H) could stunt larval growth and waste conversion due to lacking a full complement of bacteria and associated functions.
| S-EPMC7921171 | biostudies-literature
Project description:Farming insects has recently emerged as a new source of protein and lipid production. To date, research has mostly focused on food applications of insects. Focusing on nonfood potential of oil and proteins of insects, high-throughput studies of insect lipids and proteins are needed. We performed proteomics and lipidomics investigation on black soldier fly (Hermitia illucens) and blow fly (Lucilia sericata) larvae to investigate new potential and applications. We used mass spectrometry for proteomics and lipidomics analysis of control and treated larvae. Treatment was performed by incubation with a biological decomposer. We provide the list of all fatty acids with their concentration in control and treated larvae. This result showed high levels of lauric acid in black soldier fly, which could even increase after biological decomposition. Proteomics analysis showed the presence of proteins like collagen of cosmetic interest, and proteins with antimicrobial properties such as phenoloxidases and enzymatic activities, such as amylase and trypsin. Insects harbor high potential for nonfood usage as additives, antimicrobial effects, and even pharmaceuticals and cosmetics. These data open avenues for future research in pharmacological and cosmetic approaches to find new molecules of interests.
| S-EPMC6592434 | biostudies-literature
Project description:Livestock manure is an important component of agricultural organic waste, and in recent years, with the development of research on the bioconversion of manure, BSFs have been proven to be useful in the treatment of a variety of livestock wastes. In-depth research on the composition of manure and its effect on the development of BSFL is, however, very scarce. The purpose of this study was to identify the parameters that influenced the growth of BSFL that was fed fattening pig manure. The pH, moisture, and nutrients of the fattening manures (namely, nursery, growing, and finishing pig manures) were measured. To examine the influence of manure types on larval growth, 100 larvae were inoculated in 100 g of each type of manure in triplicate. According to the findings, larvae fed finishing pig manure had the lowest dry weight (30.2 ± 6.1 mg) compared to those fed growing (58.2 ± 7.3 mg) or nursery (65.5 ± 6.2 mg) pig manure. The correlation coefficients (r) between the nutrients in the manure and the weight of the larvae were calculated. Hemicellulose had the greatest |r| value (0.9569). Further research revealed that larvae raised on hemicellulase-pretreated finishing pig manure frequently weighed 21-30% (days 2-8) more than larvae raised on control manure. In conclusion, hemicellulose was a significant component that might hinder larval growth. The results of this study could be used to improve the system before it is put into use.
| S-EPMC9913757 | biostudies-literature
Project description:Hermetia illucens L., known as Black Soldier Fly (BSF) appear as an opportunity to reuse vegetable by-products, as it is easy to reproduce and can be reared in agricultural side streams, allowing the production of both, animal feed (the larvae, after recycling of the vegetal debris) and soil organic fertilizer (insect frass). Although several organic fertilizers, from long ago, have been used in agriculture, there is yet a paucity of experimental data on the evaluation of the fertilization potential of BSF larvae frass (BSFF). The present study is a contribution to access the agronomic and environmental potential of the BSFF as an organic fertilizer. Within this aim, it was conducted a greenhouse experiment with ryegrass, using seven treatments of BSFF. Under the experimental conditions, the results showed a significant effect of BSFF on the overall ryegrass production, with a steady increase (significant at p ≤ 0.05, as accessed through the Tukey test) up to the treatment with a greater rate of application. In what concerns sustainability of soil productivity, at the end of the experiment, there was also positive indications, namely, a significant increase of OM, P and K, for treatments with higher N endowments, together with a constant increase of dehydrogenase activity, from the control to the higher treatment, which was significant for treatments receiving the higher dose of BSFF.
| S-EPMC7785954 | biostudies-literature
Project description:Significant economic, environmental, and social impacts are associated with the avoidable disposal of foods worldwide. Mass-rearing of black soldier fly (Hermetia illucens) larvae using organic wastes and food- and agro-industry side products is promising for recycling resources within the food system. One current challenge of this approach is ensuring a reliable and high conversion performance of larvae with inherently variable substrates. Research has been devoted to increasing rearing performance by optimizing substrate nutrient contents and ratios, while the potential of the substrate and larval gut microbiota to increase rearing performance remains untapped. Since previous research has focused on gut microbiota, here, we describe bacterial dynamics in the residue (i.e., the mixture of frass and substrate) of black soldier fly larvae reared on two food wastes (i.e., canteen and household waste). To identify members of the substrate and residue microbiota, potentially associated with rearing performance, bacterial dynamics were also studied in the canteen waste without larvae, and after inactivation by irradiation of the initial microbiota in canteen waste. The food waste substrates had similar microbiota; both were dominated by common lactic acid bacteria. Inactivation of the canteen waste microbiota, which was dominated by Leuconostoc, Bacillus, and Staphylococcus, decreased the levels of all rearing performance indicators by 31-46% relative to canteen waste with the native microbiota. In both food waste substrates, larval rearing decreased the bacterial richness and changed the physicochemical residue properties and composition over the rearing period of 12 days, and typical members of the larval intestinal microbiota (i.e., Providencia, Dysgonomonas, Morganella, and Proteus) became more abundant, suggesting their transfer into the residue through excretions. Future studies should isolate members of these taxa and elucidate their true potential to influence black soldier fly mass-rearing performance.
| S-EPMC7719680 | biostudies-literature
Project description:Insects are a promising alternative protein source. One of the bottlenecks in applying insects in food is the fast darkening initiated during grinding. Besides enzymatic browning, non-enzymatic factors can cause off-colour formation, which differs between species. This study investigates the impact of iron, phenoloxidase, and polyphenols on off-colour formation in insect larvae. Hermetia illucens showed a blackish colour, whereas Tenebrio molitor turned brown and Alphitobius diaperinus remained the lightest. This off-colour formation appeared correlated with the iron content in the larvae, which was 61 ± 9.71, 54 ± 1.72 and 221 ± 6.07 mg/kg dw for T. molitor, A. diaperinus and H. illucens, respectively. In model systems, the formation of iron-L-3,4-dihydroxyphenylalanine (L-DOPA) bis- and tris-complexes were evidenced by direct injection into ESI-TOF-MS, based on their charges combined with iron isotope patterns. The reversibility of the binding of iron to phenolics, and thereby loss of blackening, was confirmed by EDTA addition. Besides complex formation, oxidation of L-DOPA by redox reactions with iron occurred mainly at low pH, whereas auto-oxidation of L-DOPA mainly occurred at pH 10. Tyrosinase (i.e. phenoloxidase) activity did not change complex formation. The similarity in off-colour formation between the model system and insects indicated an important role for iron-phenolic complexation in blackening.
| S-EPMC6393531 | biostudies-literature
Project description:The black soldier fly is currently the most produced edible insect on industrial scale, with its larval stage being processed into animal feed as the main application. As this insect species enters the feed and food chain, good hygiene and monitoring practices are needed to avoid the entrance of foodborne pathogens via the larvae. However, insufficient data on the risk of such introductions via industrial larvae production are available. To address this gap, a range of rearing trials were conducted in which the substrate, chicken feed, was inoculated with different levels of Salmonella and in which total viable counts and Salmonella counts were determined during the following days. The outgrowth of Salmonella was slower in those experiments with a lower initial contamination level than in experiments with a higher level. No significant reducing effect originating from the larvae on the substrate Salmonella counts was observed, in contrast to previous studies using other substrates. Our study also revealed that airborne transmission of Salmonella is possible under rearing conditions corresponding to those applied at industrial production sites. Based on our results, we recommend insect producers to use substrate ingredients free of Salmonella, and not to count on the antimicrobial activities that BSFL may exert in some situations towards food pathogens. More inoculation studies using other Salmonella serotypes, other zoonotic bacteria, other substrates, larvae of other ages and including variations on rearing protocols are needed in order to obtain a general view on the dynamics of food pathogens in this insect species and to support comprehensive risk assessments.
| S-EPMC8505792 | biostudies-literature