Brevitarsis Larvae Research Articles

Application of enzyme preparations in the treatment of sawdust spent mushrooms substrate by Protaetia brevitarsis larvae

Application of enzyme preparations in the treatment of sawdust spent mushrooms substrate by Protaetia brevitarsis larvae

Comparison of plastic and glass cages on volatile compounds in protein extracted from Protaetia brevitarsis larvae

Edible insects, known for their high-protein production efficiency, are vital for enhancing food security. However, standardized breeding protocols are lacking, and research on the impact of cage materials on insect product composition is limited. This study investigated the volatile compounds and processing properties of protein from Protaetia brevitarsis larvae reared in plastic and glass cages. Protein extracts from larvae reared in plastic cages contained 14 types of hydrocarbons, 4 types of ketones, and 1 type of phenol. Those reared in glass cages contained two types of acids, seven types of alcohols, and five types of aldehydes. Notably, plastic-derived compounds, such as p-xylene (110.87 μg/mL) and o-xylene (37.98 μg/mL), were significantly higher in the extracts from plastic cages, indicating potential plastic exposure. Processing properties, including protein solubility, pH, color, foaming properties, and emulsion characteristics, showed no significant differences between the two rearing conditions (P > 0.05). Therefore, considering the detection and potential accumulation of plastic-derived volatile compounds, using glass cages may be more beneficial for rearing insects for protein production.

Investigating the Nutritional and Functional Properties of Protaetia brevitarsis Larvae and Isolated Soy Protein Mixtures as Alternative Protein Sources.

The growing demand for sustainable and alternative protein sources has spurred interest in insect-based and plant-based proteins. Protaetia brevitarsis (PB) larvae and isolated soy protein (ISP) are notable in this regard, offering potential health benefits and nutritional enhancements. We assessed the feasibility of PB larvae and ISP mixtures as alternative food ingredients. Methods included the optimized purification and freeze-drying of PB larvae, extraction and refinement of legume proteins, physicochemical and antioxidant capacity evaluations, DPPH radical scavenging activity measurement, total phenolic and flavonoids content quantification, general component analysis, amino acid profiling using HPLC, fatty acid profiling through gas chromatography, and mineral content analysis using inductively coupled plasma spectrometry. The study found that certain PB:ISP ratios, particularly a 7:3 ratio, significantly improved the blend's antioxidant capacity, as evidenced by DPPH scavenging activity. This ratio also impacted the nutritional profile by altering the mixture's general components, with a notable increase in moisture, crude protein, and fiber and a decrease in crude fat and ash. Amino acid analysis revealed a balanced presence of essential and non-essential amino acids. The fatty acid profile was rich in unsaturated fatty acids, especially in certain ratios. Mineral analysis showed a complex interplay between PB larvae and ISP, with some minerals decreasing and others increasing in the blend. PB larvae and ISP mixtures have significant potential as alternative protein sources, offering a diversified nutritional profile and enhanced antioxidant properties. The 7:3 ratio of PB larvae to ISP has been shown to be particularly effective, suggesting that this ratio may offer an optimal balance for enhancing the overall nutritional quality of the mixture. This study sets the stage for future research to further explore and optimize the potential of these mixtures for human consumption while considering the challenges of consumer acceptance and long-term safety.

Characterization and evaluation of actinomycete from the Protaetia brevitarsis Larva Frass.

Protaetia brevitarsis larvae (PBL) are soil insects important for the soil organic carbon cycle, and PBL frass not only contains a large amount of humic acid but also affects the diversity, novelty, and potential functions of actinomycetes. Here, we characterized and assessed the actinomycete. The operational taxonomic unit (OTU) data showed that 90% of the actinomycetes cannot be annotated to species, and pure culture and genome analysis showed that 35% of the strains had the potential to be new species, indicating the novelty of PBL frass actinomycetes. Additionally, genome annotation showed that many gene clusters related to antifungal, antibacterial and insecticidal compound synthesis were identified, and confrontation culture confirmed the antifungal activities of the actinomycetes against soil-borne plant pathogenic fungi. The incubation experiment results showed that all isolates were able to thrive on media composed of straw powder and alkaline lignin. These results indicated that PBL hindgut-enriched actinomycetes could survive in soil by using the residual lignocellulose organic matter from plant residues, and the antibiotics produced not only give them a competitive advantage among soil microflora but also have a certain inhibitory effect on plant diseases and pests. This study suggests that the application of PBL frass can not only supplement soil humic acid but also potentially affect the soil microbiota of cultivated land, which is beneficial for the healthy growth of crops.

Variation of volatile compounds and sensory profile for Protaetia brevitarsis larvae fermented with lactic acid bacteria and yeast

This study aimed to investigate the correlation between the composition of volatile compounds, consumer acceptance, and drivers of (dis)liking of Protaetia brevitarsis larvae fermented using lactic acid bacteria and yeast. Volatile compounds were analyzed using HS-SPME-Arrow-GC–MS, and a sensory evaluation was conducted with 72 consumers. A total of 113 volatile compounds were detected, and principal component analysis indicated that the samples could be divided into three groups. The calculated relative odor activity values (ROAV) revealed the presence of 27 compounds (ROAV >1). Volatile compounds with high ROAV were predominantly found during yeast fermentation. The sensory evaluation results indicated a strong correlation between low levels of off-odor intensity and high odor liking, emphasizing that odor profile had a more direct association with consumer acceptance than odor intensity. These findings suggest that yeast fermentation using volatile compounds, which positively influences consumer acceptance, is appropriate for Protaetia brevitarsis larvae.

Protaetia brevitarsis larvae produce frass that can be used as an additive to immobilize Cd and improve fertility in alkaline soils

Bioconversion of agricultural waste by Protaetia brevitarsis larvae (PBL) holds significant promise for producing high-quality frass organic amendments. However, the effects and mechanisms of PBL frass on Cd immobilization in an alkaline environment remain poorly understood. In this study, three types of frass, namely maize straw frass (MF), rice straw frass (RF), and sawdust frass (SF), were produced by feeding PBL. The Cd immobilization efficiencies of three frass in alkaline solutions and soils were investigated through batch sorption and incubation experiments, and spectroscopic techniques were employed to elucidate the sorption mechanisms of Cd onto different frass at the molecular level. The results showed that MF proved to be an efficient sorbent for Cd in alkaline solutions (176.67–227.27 mg g−1). X-ray absorption near-edge structure (XANES) spectroscopy indicated that Cd immobilization in frass is primarily attributed to the association with organic matter (OM-Cd, 78–90%). And MF had more oxygen-containing functional groups than the other frass. In weakly alkaline soils, MF application (0.5–1.5%) significantly decreased Cd bioavailability (5.65–18.48%) and concurrently improved soil nutrients (2.21–56.79%). Redundancy analysis (RDA) unveiled that pH, CEC, and available P were important factors controlling Cd fractions. Path analysis demonstrated that MF application affected Cd bioavailability directly and indirectly by influencing soil chemical properties and nutrients. In summary, MF, the product of PBL-mediated conversion maize straw, demonstrated promise as an effective organic amendment for Cd immobilization and fertility improvement in alkaline soils.

Comparative Analysis of Hindgut Microbiota Variation in Protaetia brevitarsis Larvae across Diverse Farms.

Protaetia brevitarsis larvae are farm-raised for food, are used in traditional East Asian medicine, and convert organic waste into biofertilizers. Here, the comparative analysis of the gut microbiota of third-instar larvae obtained from five different farms was investigated using 16S rRNA microbial profiling. Species richness, evenness, and diversity results using α-diversity analysis (observed species, Chao1, Shannon, Simpson) were similar between farms, except for those between the TO and KO farms. β-diversity was significantly different in distribution and relative abundance between farms (PERMANOVA, pseudo-F = 13.20, p = 0.001). At the phylum level, Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota were the most dominant, accounting for 73-88% of the hindgut microbial community. At the genus level, Tuberibacillus, Proteiniphilum, Desulfovibrio, Luoshenia, and Thermoactinomyces were the most abundant. Although oak sawdust was the main feed component, there were large variations in distribution and relative abundance across farms at the phylum and genus levels. Venn diagram and linear discriminant analysis effect size analyses revealed large variations in the hindgut microbial communities of P. brevitarsis larvae between farms. These results suggest environmental factors were more important than feed ingredients or genetic predisposition for the establishment of the intestinal microbiota of P. brevitarsis larvae. These findings serve as reference data to understand the intestinal microbiota of P. brevitarsis larvae.

Fermented Protaetia brevitarsis Larvae Improves Neurotoxicity in Chronic Ethanol-Induced-Dementia Mice via Suppressing AKT and NF-κB Signaling Pathway.

This study was investigated to examine the neuroprotective effect of fermented Protaetia brevitarsis larvae (FPB) in ethanol-induced-dementia mice. Consumption of FPB by mice resulted in improved memory dysfunction in the Y-maze, passive avoidance, and Morris water maze tests. FPB significantly decreased oxidative stress by regulating levels of malondialdehyde (MDA), superoxide dismutase (SOD), and reduced glutathione (GSH) in brain tissues. In addition, FPB restored cerebral mitochondrial dysfunction by modulating levels of reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and ATP. In addition, FPB enhanced the cholinergic system via the regulation of acetylcholine (ACh) content, acetylcholinesterase (AChE) activity, and expressions of AChE and choline acetyltransferase (ChAT) in brain tissues. FPB ameliorated neuronal apoptosis through modulation of the protein kinase B (AKT)/B-cell lymphoma (BCL)-2 signaling pathway. Also, FPB improved inflammation response by down-regulating the toll-like receptor (TLR)-4/nuclear factor (NF)-κB pathway. Additionally, FPB ameliorated synaptic plasticity via the increase of the expressions of synaptophysin (SYP), postsynaptic density protein (PSD)-95, and growth-associated protein (GAP)-43. Treatment with FPB also reinforced the blood-brain barrier by increasing tight junctions including zonula occludens (ZO)-1, occludin, and claudin-1. In conclusion, these results show that FPB can improve cognitive impairment via AKT/NF-κB pathways in ethanol-induced-dementia mice.

Antithrombotic effects of Protaetia brevitarsis seulensis larvae in a rat tail thrombosis model using κ‐carrageenan

AbstractThe present study aimed to investigate the preventive role of Protaetia brevitarsis seulensis (P. brevitarsis) larvae in the κ‐carrageenan‐induced rat tail thrombosis model, in comparison with that of heparin. Thrombosis was induced in Sprague–Dawley (SD) rats by intraperitoneal injection with a dose of 30 mg/kg κ‐carrageenan after receiving oral pretreatment with P. brevitarsis larvae at doses of 300 or 900 mg/kg. The control group was only given physiological saline for 7 days without the administration of κ‐carrageenan. The results indicate that pretreatment with P. brevitarsis larvae at 300 or 900 mg/kg not only significantly reduced the average length of the thrombus but also markedly decreased the fibrinogen levels and serum P‐selectin in a dose‐dependent manner (P < 0.05). However, there were no notable effects on the blood clotting times, such as prothrombin time (PT) and activated partial thromboplastin time (aPTT). Moreover, the histological analysis of tail tissues treated with P. brevitarsis larvae supported the serum biochemical findings, thereby providing evidence of enhanced blood circulation. Based on these results, it was concluded that P. brevitarsis larvae had a positive effect on preventing thrombosis in vivo through the reduction of fibrinogen and P‐selectin levels, suggesting potential applications for thrombosis prevention.

Development of feed material and its effect on the nutritional composition of Protaetia brevitarsis larvae

AbstractThis study was carried out to assess the nutritional composition of Protaetia brevitarsis larvae (Pbl) reared on different feed materials. Four different types of feed materials were used to rear Pbl prior to nutritional analysis. The results show that the new formulation of feed material enhanced the crude fat content of Pbl by 75%, compared with the control. A marked increase in unsaturated fatty acids such as linoleic acid and oleic acid, by 100% compared with the control, in Pbl reared on the new formulation of feed material was also observed.

Parameter calibration and experimental verification of discrete element simulation model for Protaetia brevitarsis larvae bioconversion mixture

Parameter calibration and experimental verification of discrete element simulation model for Protaetia brevitarsis larvae bioconversion mixture

Protaetia brevitarsis larvae extract protects against lipopolysaccharides-induced ferroptosis and inflammation by inhibiting acid sphingomyelinase.

Inflammation and ferroptosis are implicated in various diseases and lipopolysaccharides (LPS) have been linked with these disorders. Recently, many edible insects, such as Gryllus bimaculatus, Protaetia brevitarsis larvae (PB) and Tenebrio molitor larvae, have been recommended as alternative foods because they contain lots of nutritional sources. In this study, we explored the potential of PB extract in preventing LPS-induced inflammation and ferroptosis in Hep3B cells. PB powder was extracted using 70% ethanol and applied to Hep3B cells. Co-treatment with LPS was conducted to induce ferroptosis and inflammation. The anti-inflammatory and anti-ferroptosis mechanisms of the PB extract were confirmed using Western blot, enzyme-linked immunosorbent assay, and real-time polymerase chain reaction analysis. PB extract effectively prevented LPS-induced cell death and restored LPS-induced inflammatory cytokine production, NF-κB signaling, endoplasmic reticulum (ER) stress and ferroptosis. Interestingly, PB extract reduced LPS-induced ceramide increase and acid sphingomyelinase (ASMase) expression. The use of the ASMase inhibitor, desipramine, also demonstrated a reduction in these pathways, highlighting the pivotal role of ASMase in inflammation and ferroptosis. Treatment with each inhibitor revealed that ferroptosis causes ER stress and that NF-κB and MAP kinase pathways are involved in inflammation. PB emerges as a potential functional food with inhibitory effects on LPS-induced inflammation and ferroptosis, making it a promising candidate for nutritional interventions.

Identification, characterization and hypolipidemic effect of novel peptides in protein hydrolysate from Protaetia brevitarsis larvae

The proteins were mainly derived from Protaetia brevitarsis larval extracts obtained using two empty intestine methods (traditional static method: TSM or salt immersion stress method: SISM) and extraction solvents (water: W or 50 % water–ethanol: W:E), and the proteins were used as objects to investigate the effect of emptying intestine methods on hypolipidemic peptides. The results revealed that the F-2 fractions of protein hydrolysate had stronger in vitro hypolipidemic activity, with the peptides obtained by SISM possessing a stronger cholesterol micelle solubility inhibition rate, especially in SISM-W:E-P. Moreover, a total of 106 peptides were tentatively identified, among which SISM identified more peptides with an amino acid number < 8. Meanwhile, five novel peptides (YPPFH, YPGFGK, KYPF, SPLPGPR and VPPP) exhibited good hypolipidemic activity in vitro and in vivo, among which YPPFH, VPPP and KYPF had strong inhibitory activities on pancreatic lipase (PL) and cholesteryl esterase (CE), and KYPF, SPLPGPR and VPPP could significantly reduce the TG content in Caenorhabditis elegans. Thus, P. brevitarsis can be developed as a naturally derived hypolipidemic component for the development and application in functional foods.

Fermented Protaetia brevitarsis Larvae Ameliorates Chronic Ethanol-Induced Hepatotoxicity in Mice via AMPK and TLR-4/TGF-β1 Pathways.

This study evaluated the hepatoprotective effect of fermented Protaetia brevitarsis larvae (FPB) in ethanol-induced liver injury mice. As a result of amino acids in FPB, 18 types of amino acids including essential amino acids were identified. In the results of in vitro tests, FPB increased alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities. In addition, FPB treatment increased cell viability on ethanol- and H2O2-induced HepG2 cells. FPB ameliorated serum biomarkers related to hepatoxicity including glutamic oxaloacetic transaminase, glutamine pyruvic transaminase, total bilirubin, and lactate dehydrogenase and lipid metabolism including triglyceride, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol. Also, FPB controlled ethanol metabolism enzymes by regulating the protein expression levels of ADH, ALDH, and cytochrome P450 2E1 in liver tissue. FPB protected hepatic oxidative stress by improving malondialdehyde content, reduced glutathione, and superoxide dismutase levels. In addition, FPB reversed mitochondrial dysfunction by regulating reactive oxygen species production, mitochondrial membrane potential, and ATP levels. FPB protected ethanol-induced apoptosis, fatty liver, and hepatic inflammation through p-AMP-activated protein kinase and TLR-4/NF-κB signaling pathways. Furthermore, FPB prevented hepatic fibrosis by decreasing TGF-β1/Smad pathway. In summary, these results suggest that FPB might be a potential prophylactic agent for the treatment of alcoholic liver disease via preventing liver injury such as fatty liver, hepatic inflammation due to chronic ethanol-induced oxidative stress.

Evaluation of Gelation Properties of Salt-Soluble Proteins Extracted from Protaetia brevitarsis Larvae and Tenebrio molitor Larvae and Application to Pork Myofibrillar Protein Gel System.

The purpose of this study was to investigate the functional properties of salt-soluble proteins obtained from Protaetia brevitarsis (PB) and Tenebrio molitor (TM) larvae, the interaction between these proteins and pork myofibrillar protein (MP) in a gel system. The gel properties of salt-soluble protein extracts showed that the PB had a higher viscosity than the TM protein. However, the TM protein had higher gel strength compared with the PB protein. The gelation characteristics of the pork MP gel systems added with lyophilized insect salt-soluble protein powder showed to decrease slightly viscosity compared with MP alone. Adding the TM or PB protein powder did not affect the pork MP's hydrophobicity and sulfhydryl group levels. Furthermore, the protein bands of the MP did not change with the type or amount of insect salt-soluble protein. The cooking yields of the pork MP gels containing PB or TM protein powder were higher than those without insect protein. Regardless of the type of insect salt-soluble protein added, the pork MP's gel strength decreased. Furthermore, as the level of insect powder increased, the surface protein structure became rough and porous. The results demonstrated that proteins extracted from PB and TM larvae interfered with the gelation of pork MP in a gel system.

Effect of gamma-ray and electron-beam irradiation on the structural changes and functional properties of edible insect proteins from Protaetia brevitarsis larvae

Edible insects are regarded as future food sources to replace traditional livestock proteins. However, insect proteins have poor processing properties owing to various structural limitations. We investigated the structure of Protaetia brevitarsis larvae proteins modified by irradiation, and analyzed their resulting processing abilities. Following irradiation with gamma rays and electron beams, the ratio of parallel β-sheets to β-turns changed significantly, and changes in the tertiary protein structures were also confirmed. The polydispersity indices of the proteins remained relatively constant following irradiation, although the zeta potential and mean diameter changed. Furthermore, the pH, protein solubility, surface hydrophobicity, foaming capacity, and emulsion stability were higher than those of the control, whereas the viscosity and foaming stability were lower. Thus, gamma ray and electron beam irradiation clearly lead to changes in the structures of edible insect proteins and improves their processing properties, promoting the industrial utilization of such proteins in the food industry.

Aqueous extract of Protaetia brevitarsis larvae increases mTOR-mediated growth rate in zebrafish larvae

Objective: To evaluate the effects of an aqueous extract of Protaetia brevitarsis (AEPB) on the growth of zebrafish and preosteoblast MC3T3-E1 cells. Methods: The effects of AEPB on the linear growth and the expression of growth-related genes in zebrafish and MC3T3-E1 cells were assessed using various molecular techniques. Furthermore, the involvement of the mammalian target of rapamycin (mTOR) pathway in AEPB-induced growth was investigated by employing the mTOR inhibitor rapamycin. Results: AEPB administration led to a significant and dose-dependent increase in zebrafish larvae growth over time. Additionally, AEPB treatment upregulated the expression of growth hormone-1 (GH-1), insulin-like growth factor-1 (IGF-1), growth hormone receptor-1 (GHR-1), and cholecystokinin-a (CCKA) in zebrafish. Similarly, AEPB stimulated the expression and release of IGF-1 and accelerated mTOR expression in MC3T3-E1 cells. In addition, rapamycin hindered AEPB-induced linear growth in zebrafish larvae and suppressed the expression of growth-promoting genes by inhibiting mTOR activation. Conclusions: AEPB shows growth-promoting effects by upregulating growth-related genes and activating the mTOR signaling pathway. Further investigations are warranted to elucidate its mechanisms of action and explore its potential application in the development of growth-enhancing supplements for various purposes.

Protaetia brevitarsis seulensis larvae ethanol extract inhibits RANKL-stimulated osteoclastogenesis and ameliorates bone loss in ovariectomized mice

Modulation of osteoclast formation could be a therapeutic target for inhibiting pathological bone destruction. The receptor activator of nuclear factor (NF)-κB ligand (RANKL) is known to be an essential factor in osteoclast differentiation and activation inducers. However, whether Protaetia brevitarsis seulensis (P. brevitarsis) larvae—a traditional animal-derived medicine used in many Asian countries—can inhibit RANKL-induced osteoclast formation and prevent ovariectomy (OVX)-induced bone loss has not been evaluated. Here, we aimed to investigate the anti-osteoporotic effects of P. brevitarsis larvae ethanol extract (PBE) in RANKL-stimulated RAW264.7 cells and OVX mice. In vitro, PBE (0.1, 0.5, 1, and 2 mg/mL) decreased RANKL‑induced tartrate-resistant acid phosphatase (TRAP) activity and expression of osteoclastogenesis-associated genes and proteins. Furthermore, PBE (0.1, 0.5, 1, and 2 mg/mL) significantly inhibited the phosphorylation of p38 and NF-κB. Female C3H/HeN mice were divided into five groups (n = 5 per group), namely, sham-operated, OVX, OVX+PBEL (100 mg/kg, oral gavage), OVX+PBEH (200 mg/kg, oral gavage), and OVX+estradiol (0.03 μg/day, subcutaneous injection). High doses of PBE significantly increased femoral bone mineral density (BMD) and bone volume/tissue volume (BV/TV), whereas femoral bone surface/bone volume (BS/BV) and osteoclastogenesis-associated protein expression decreased compared to those in the OVX group. Moreover, PBE (200 mg/kg) significantly increased estradiol and procollagen type I N-terminal propeptide and decreased N-terminal telopeptide of type I collagen and C-terminal telopeptide of type I collagen compared to those in the OVX group. Our results suggest that PBE can be an effective therapeutic candidate for preventing or treating postmenopausal osteoporosis.

Structural, physicochemical, and immune-enhancing properties of edible insect protein isolates from Protaetia brevitarsis larvae.

Edible insects are promising future food resources globally. Herein, the structural, physicochemical, and bio-functional properties of edible insect protein isolates (EPIs) extracted from Protaetia brevitarsis larvae were investigated. The results showed that EPIs have a high total essential amino acid content; moreover, β-sheet is the major secondary protein structure. The EPI protein solution was highly soluble and electrically stable and did not aggregate easily. In addition, EPIs exhibited immune-enhancing properties; EPI treatment of macrophages induced the activation of macrophages and consequently promoted the production of pro-inflammatory mediators (NO, TNF-α, and IL-1β). Moreover, macrophage activation of EPIs was confirmed to occur through the MAPK and NF-κB pathways. In conclusion, our results suggest that the isolated P. brevitarsis protein can be fully utilized as a functional food material and alternative protein source in the future food industry.

White-spotted flower chafer (Protaetia brevitarsis) ameliorates inflammatory responses in LPS-stimulated RAW 264.7 macrophages

White-spotted flower chafer (Protaetia brevitarsis) is an edible insect andP. brevitarsis larvae are used in traditional Oriental medicine. However, the molecular mechanisms behind the anti-inflammatory action ofP. brevitarsis larvae are unknown. We investigated the effects ofP. brevitarsis larvae ethanol extract (PBLE) on the inflammatory response as well as the molecular mechanisms underpinning this effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. PBLE effectively inhibited the synthesis of nitric oxide (NO) through inducible NO synthase (iNOS) and cyclooxygenase 2 protein expression and 20 μM PBLE suppressed the expression of pro-inflammatory cytokines, including interleukin (IL)-6 and IL-8, at a dose of. Further investigation of the mechanisms revealed that PBLE attenuated phosphorylation of nuclear factor kappa B (NF-κB) inhibitor alpha (IκB-α) and NF-κB p65 and interfered selectively with extracellular signal-regulated kinase (ERK)1/2 and p38 phosphorylation. These findings provide the first molecular basis that PBLE could be a potential candidate for attenuating proinflammatory NO induction via selective inhibition of the NF-κB, ERK, and p38 mitogen-activated protein kinase signalling pathways. It is necessary to verify the anti-inflammatory action of PBLE and to ascertain the related mechanisms, both inin vivo models and in clinical studies, to enable the application of PBLE as a potential therapeutic agent for inflammatory diseases.