Treatment of biowaste commingled with biodegradable bioplastic films using Black Soldier Fly larvae: Generation and fate of micro-plastics.

The use of Black Soldier Fly (BSF) larvae is emerging as a promising alternative for biowaste (i.e. food waste) treatment, generating larval biomass and process residues, suitable for use as animal feed and fertilizer, respectively. In line with an increasing use of starch-based bioplastics in food packaging, the presence of these biopolymers and associated biodegradable microplastics (BMPs) in food waste is expected to rise. Knowledge of the generation of BMPs and their fate in the BSF treatment process is scarce, or indeed, completely lacking in the case of small-sized BMPs (<50 µm). The present study aims to investigate the generation and potential accumulation of BMPs in BSF larvae process. Food waste mixed with starch-based bioplastic films was fed to larvae and BMPs of two particle sizes (inferior to and exceeding 10 µm in diameter) were monitored over time in rearing substrate and larval biomass. BMPs concentrations in substrate were compared with larvae-free control tests. The presence of larvae favoured the generation of BMPs. Concentrations of smaller-sized BMPs (<10 µm) increased by approximately 172% in the final substrate, and accumulated in the larval biomass with a peak exceeding the initial larval concentration by over 1000% just before prepupation, which is the typical stage they are collected when used as animal feed. These results indicate a potential risk of soil contamination by BMPs when final substrate is used as fertilizer and a risk of biomagnification phenomena when larvae are used as animal feed.

Structural characterization, HepG2 cell cytoprotective ability, and antioxidant mechanism of novel antioxidant peptides identified from black soldier fly larvae (Hermetia illucens L.).

This study isolated a novel antioxidant peptide from black soldier fly larvae (BSFL) using enzymatic hydrolysis. Firstly, the BSFL enzymatic hydrolysate was fractionated through ultrafiltration, with the <3 kDa fraction exhibiting the strongest DPPH and ABTS radical scavenging activity. Subsequently, this fraction was further fractionated through gel filtration chromatography and RP-HPLC. Totally, 153 peptides were identified through LC-MS/MS analysis, from which a novel peptide EDEGTYKCVLS (Pep6) was screened according to activity prediction and verification. Pep6 exhibited high radical scavenging capacity and cytoprotective effect on HepG2 cells against H2O2 damage, meanwhile significantly increasing the intracellular antioxidant enzymes activity. Molecular docking analysis indicated that Pep6 competitively bound to Keap1, thereby inhibiting the formation of Keap1-Nrf2 complex, ultimately protecting cells from oxidative stress damage. In this study, a novel antioxidant peptide Pep6 was identified from BSFL, and its antioxidant mechanism was elucidated, providing a theoretical basis for its use as a natural antioxidant.

A new and widely distributed species of Ichthyurus Westwood, 1848 (Cantharidae, Chauliognathinae, Ichthyurini) from China.

Background: The genus Ichthyurus Westwood, 1848 is a large cantharid group consisting of approximately 200 species worldwide, with only 10 species hitherto found in China. Despite its expansive area, the Chinese fauna has historically received little attention from specialists, leading to a lack of knowledge even about some common Ichthyurus species in this region. New information: A new species of Ichthyurus Westwood, 1848 is described under the name of I.longulus sp. nov., which is widely distributed in mainland China, including Shannxi, Gansu, Hubei, Chongqing, Guizhou and Guangxi. Although there are some variations in the pronotum colouration within the species, this new species could be easily distinguished from all others of Ichthyurus by the large-sized body, uniformly black elytra, mesotibiae each with an apical spur in male, terminal abdominal tergite of male with long and cylindrical lateral projections that are about 3/5 the length of the tergite, terminal abdominal ventrite of male saddle-shaped and deeply cleft in middle of apical 2/3 part and aedeagus with a long setifore extension that is as long as the parameres. The habitus, terminal abdominal ventrite and tergites and genitalia of both sexes are illustrated. In addition, a distribution map of this species and a list of the Ichthyurus species from mainland China are provided.

Effects of heat treatment on rumen degradability and protein intestinal digestibility of black soldier fly (Hermetia illucens L.) in goat.

The black soldier fly larvae (BSF) are used as a substitute for soybean meal due to their high crude protein content. This experiment aims to assess the impact of heat treatment on the rumen degradability of BSF and protein digestion in the small intestine using the in situ nylon bag method and the three-step in vitro method. This study comprises a total of 8 groups (n = 6). The negative control group includes only full-fat soybeans (FFS) and BSF (FF group and BS group). The positive control groups consist of a 95% BSF or 95% FFS mixed with 5% cassava (FFC and BSC groups). The treatment groups involve adding 75% water to the positive control mixture, followed by vigorous kneading to achieve uniform mixing. The resulting mixture was then pressed to a thickness of approximately 5 cm, placed in an oven, and dried for 120 min at temperatures of 120 °C and 140 °C (12FFC, 14FFC, 12BSC, and 14BSC groups). Nylon bags will be incubated in the rumen for 0, 2, 4, 8, 12, 24, and 48 h, and the small intestine protein digestion rate will be analyzed at 16 h. Compared to the BS group, heat-treated BSF showed increased (P < 0.05) rumen DM degradability and effective degradability. The 14BSC group increased (P < 0.05) rumen CP degradability and degradation kinetic parameters, while the 12BSC group decreased (P < 0.05) these parameters. The CP degradability of BSF was significantly higher (P < 0.05) than that of full-fat soybeans. The Idg and IDCP of heat-treated full-fat soybeans were significantly higher (P < 0.05) than those of other treatment groups. At the same time, heat treatment was beneficial for increasing (P < 0.05) the Idg and IDCP of BSF, and the 14BSC treatment effect was significantly better (P < 0.05) than that of the 12BSC group. Therefore, based on the results of this experiment, it is recommended to supplement BSF with cassava and subject them to heat treatment at 140 °C.

Effects of black soldier fly (Hermetia illucens) larvae meal on production performance, egg quality, and physiological properties in laying hens: A meta-analysis.

Background and Aim: The primary components of fat and protein in chicken diets are fishmeal and soybean; however, due to limited supply and high costs, several efforts have been made to utilize alternative feedstuffs. The potential of black soldier fly larvae (BSFL) as a substitute for fat and protein has been extensively studied, but the findings are not consistent. This study used a meta-analysis approach to investigate the integrated efficacy of BSFL supplementation on laying hen production performance, egg quality, and physiological properties. Materials and Methods: The articles were retrieved from PubMed, Scopus, ScienceDirect, Cochrane Library, and ProQuest. The retrieved references were examined for potential inclusion. The relevant findings of the included studies were then extracted. Fixed-effects, standard mean difference, 95% confidence intervals, and heterogeneity models were analyzed using the Review Manager website version (Cochrane Collaboration, UK). Results: A total of 24 papers from 17 different nations across five continents have been selected for meta-analysis out of the 3621 articles that were reviewed. The current meta-analysis demonstrated that providing BSFL meals significantly favored feed efficiency, haugh units, albumen quality, eggshell quality, serum glucose, and lipid levels. In addition, significant trends in alanine transaminase, alkaline phosphatase, magnesium, phosphorus, chlorine, and iron levels were observed in blood urea nitrogen, uric acid, creatinine, lactate dehydrogenase, creatine kinase, glutathione peroxidase, and malondialdehyde. On the other hand, it was revealed that there was no favorable effect on weight gain, laying, yolk quality, and hematological profile. Conclusion: The meta-analysis confirmed that BSFL meals can be utilized to optimize feed efficiency, haugh units, albumen, eggshell quality, liver, renal, and cellular physiology of laying hens, although they did not significantly increase body weight gain, laying production, and hematological profiles.

The Growth Performance and Nutrient Composition of Black Soldier Fly (Hermetia illucens) Larvae Fed Slaughtered Bovine Blood.

The disposal of slaughterhouse blood poses significant environmental challenges due to its biological instability and high nutrient content. We used a gradient of 10% blood increments (0-100%) to feed BSFL, and the correlation between the proportion of bovine blood and the BSFL weight gain, mortality rate, fatty acid content, and amino acid content was researched. Results indicate a positive correlation between the bovine blood content and BSFL mortality, with survival rates above 95% for blood proportions below 60%. Larval weight exhibited a negative correlation as the bovine blood content increased. Nutritional analysis revealed that the crude protein content in BSFL increased proportionally with bovine blood (14.75-25.45 g/100 g), while the crude fat content decreased correspondingly (10.70-4.66 g/100 g). The sugar content remained relatively constant across groups. Fatty acid analysis showed increased levels of C16:0, C14:0, and C16:1 and decreased levels of C18:1, C18:2, and C18:3 with higher bovine blood contents. The amino acid content generally increased with higher blood proportions. This study highlights the bioconversion potential of BSFL for bovine blood and underscores the impact of protein, lipid, and sugar concentrations in feed on BSFL growth. These findings provide valuable insights for utilizing slaughterhouse waste in BSFL rearing, contributing to the development of more sustainable waste management and animal feed production methods.

The impact of early-life conditions on visual discrimination abilities in free-ranging laying hens.

Conditions during incubation and rearing can greatly affect the developmental trajectory of chickens, in a positive and negative way. In this study, the effect of early-life conditions on the visual discrimination abilities of adult, free-ranging laying hens was examined. These early-life treatments entailed incubation in a 12/12h green light/dark cycle and rearing with Black soldier fly larvae (BSFL) as foraging enrichment. Through a modified pebble-floor test, 171 hens of 41 to 42 wk old, housed in mobile stables with outdoor access, were tested for their ability to discriminate between food and nonfood items (mealworms and decoy mealworms). Each hen was allowed 60 pecks during the trial, from which the overall success rate, as well as within-trial learning was investigated. The latter was accomplished by dividing the 60 pecks into 3 blocks of 20 pecks and comparing the success rate between these blocks. Due to another ongoing experiment on range use, roughly half the hens received range enrichment (mealworms) at the time of testing, so this was included as a covariate in the analysis. Incubation with green light did not have an effect on the visual discrimination abilities of adult laying hens. Rearing with BSFL did have a limited beneficial effect on the visual discrimination abilities, as evidenced by a higher success rate during the first block of the visual discrimination trial. These enhanced visual discrimination abilities might be useful in a more complex free-range setting, where the animals have more foraging opportunities. Hens that received range enrichment at the time of testing, also had a higher success rate during the visual discrimination test, though they had a lower degree of test completion, likely due to habituation to the mealworms as an enrichment. The positive effects of BSFL during rearing and mealworms during the laying period stress the importance of enrichment throughout the life of the hens.

The genome sequence of a soldier beetle, Malthinus flaveolus (Herbst, 1786).

We present a genome assembly from an individual female Malthinus flaveolus (soldier beetle; Arthropoda; Insecta; Coleoptera; Cantharidae). The genome sequence is 236.7 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 19.27 kilobases in length. Gene annotation of this assembly on Ensembl identified 16,617 protein coding genes.

Assessing the performance, egg quality, serum analysis, heavy metals and essential trace metals accumulation in laying hen eggs and tissues fed black soldier fly (Hermetia illucens) larvae meal.

Black soldier fly (BSF) larvae convert wastes into protein, playing a vital role in addressing the challenge of sustainable poultry production. These larvae accumulate toxic substances, posing a risk to feed and food safety. This study investigates the effects of substituting soybean meal with different levels of BSF larvae meal on laying performance, egg quality, serum analysis, and the deposition of various metals in eggs, meat, and excreta. A total of 1,008 Lohmann hens (age 48 wk) were randomly assigned to 4 treatments. The treatments consisted of corn-soybean meal (CK) diet replaced with 7% (BSF7), 14% (BSF14), and 21% (BSF21) BSF larvae meal. Each treatment consisted of 6 replicates with 42 hens each and the trial lasted for 8 wk. Dietary BSF larvae meal treatments increased (linear, P < 0.05) the laying rate (1.52 to 1.95%) and decreased (linear, P < 0.01) the feed intake (3.64-3.86 g) during the entire experiment. During 48 to 52 wk, egg weight was decreased (P < 0.001) 0.93 g for the BSF21 group compared to CK group, however, during 52-56 wk, no differences in egg weight were observed among treatments. The addition of BSF larvae meal enhanced the eggshell strength (linear, P < 0.05), with no effect on the albumen height and yolk weight (P > 0.05). Low transfer of arsenic, lead, and cadmium concentration was observed in the egg yolk and egg white across different treatments (P > 0.05). Conversely, the concentrations of these metals and iron showed an increase, while Zinc exhibited a decrease in excreta as the dietary intake of BSF larvae meal increased. The chromium and iron increased linearly (P < 0.001) in the egg white with the inclusion of BSF larvae meal. Egg white iron and chromium showed a strong positive correlation with the dietary zinc and copper. Taken together, BSF larvae can replace soybean meal completely in laying diet, however, careful attention requires to elevated metal levels in diet and excreta.

Exploring the potential of Hermetia illucens larvae extracts: A promising approach for dermocosmetic formulations.

Globally, the yearly disposal of 1.3 billion tonnes of food raises environmental and public health concerns. Black soldier fly (BSF) larvae present a sustainable solution, converting organic waste into nutrient-rich biomass. The extracted oil from BSF larvae, rich in fatty acids (FA), offers an eco-friendly alternative for the cosmetic industry. In this study, larvae sourced from a Portuguese company were fed olive pomace, a by-product of olive oil production. The lipidic sample extracted revealed a composition high in oleic acid, valuable for cosmetics. Investigating the biological activity of lipid extractions from larvae fed with olive pomace is a novel approach. Notably, the n-hexane ultrasound-assisted extraction method demonstrated potent antioxidant properties, and some extracts displayed antimicrobial activity. Five non-cytotoxic extracts; three with no relevant activity (IC50 from 236 to >400 µg/mL). These findings highlight BSF larvae as an environmentally friendly source of fatty acids, offering promising alternatives for diverse applications.

Screening of oxytetracycline-degrading strains in the intestine of the black soldier fly larvae and their degradation characteristics.

The presence of excessive antibiotic residues poses a significant threat to human health and the environment. This study was designed to identify an effective oxytetracycline (OTC)-degrading strain through the screening of the intestine of black soldier fly larvae (BSFL). A strain designated "B2" was selected using a series of traditional microbial screening methods. It could be identified as Enterococcus faecalis by Gram staining and 16S rDNA sequencing, with a similarity of 99.93%. Its ability to degrade OTC was then assessed using high-performance liquid chromatography (HPLC). The degradation of the strain was characterized using a one-way test to assess the effects of the substrate concentration, inoculum amount, and initial pH on the degrading bacteria. The results indicate that strain B2 exhibited optimal OTC-degrading performance at a substrate concentration of 50 mg/L, with an inoculum amount of 6% and a pH value of 5.0. Specifically, strain B2 achieved degradation rates of 71.11%, 56.14%, and 45.03%. These findings demonstrate the effectiveness of strain B2 in degrading OTC, indicating its potential for use in environmental remediation efforts.

Unraveling the role of black soldier fly larvae in chicken manure conversion: Facilitating maturation and enhancing humification.

Black soldier fly larvae (BSFL) have garnered considerable attention for their efficacy in mitigating waste management challenges. However, their potential in treating antibiotics contaminated chicken manure remains uncertain. This study investigates the physicochemical properties changes and nutrient dynamics during the composting of contaminated-chicken manure using BSFL. The results indicate that BSFL treatment reduces electrical conductivity (by 6.01-58.09 %), organic matter, and dissolved organic carbon content in chicken manure throughout the composting process, while maintaining a more stable pH value (pH âˆ¼ 6.0-8.0). This is attributed to the consumption of organic matter by BSFL and the subsequent promotion of organic acid formation. Additionally, BSFL treatment improves the degree of aromatization of dissolved organic matter (DOM) in chicken manure and increases the proportions of fulvic acid (up to 48.77 %) and humic acid (maximally 14.27 %) within the DOM. The germination index and pot experiments indicated improved compost maturity and plant growth in BSFL-treated composts. Furthermore, BSFL meal demonstrated high protein and essential fatty acid content, highlighting its potential as a protein supplement in animal feed. This study underscores the efficacy of BSFL in enhancing compost quality and nutrient availability, offering a sustainable solution for waste management and animal feed production.

The genome sequence of a soldier beetle, Cantharis flavilabris Fallén, 1807.

We present a genome assembly from an individual female Cantharis flavilabris (soldier beetle; Arthropoda; Insecta; Coleoptera; Cantharidae). The genome sequence is 348.3 megabases in span. Most of the assembly is scaffolded into 7 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 17.5 kilobases in length. Gene annotation of this assembly on Ensembl identified 22,711 protein coding genes.

Dietary black soldier fly oil enhances growth performance, flesh quality, and health status of largemouth bass (Micropterus salmoides).

The study aimed to assess the effects of dietary black soldier fly oil (BSFO) on the growth performance, flesh quality, and health status of largemouth bass (Micropterus salmoides). Six iso-nitrogenous and isolipid diets were formulated by substituting fish oil and soybean oil (1/2, wt/wt) with BSFO in percentages of 0%, 20%, 40%, 60%, 80%, and 100%, respectively. The diets were fed to 960 fish (initial body weight = 16.5 g) in four replicates for 8 weeks. Indicators related to growth performance, body composition, hematology, flesh quality, expression of genes related to inflammatory cytokines and apoptosis, and the response of fish to Aeromonas veronii challenge were analyzed. The results showed that the weight gain rate was numerically improved in all BSFO substitution groups, ranging from 9.3% to 44.0% compared to the control group. The highest survival rate and the lowest hepatosomatic index and condition factor were observed in the BFSO20 group. In terms of flesh quality, the water-holding capacity of the dorsal muscle was elevated with higher levels of dietary BSFO. However, significant changes in texture properties (cohesiveness, gluing, and chewiness) were observed in the BSFO20 group (P < 0.05). Six hematological parameters related to glycolipid and liver function were optimized in most of the BFSO substitution groups. Furthermore, the expressions of six inflammation- and apoptosis-related genes (IL-1ß, Bcl-xl, BAX, caspase8, TNF-α, and IL-10) were significantly affected by dietary BSFO (P < 0.05). Following bacterial challenge, the seven-day cumulative survival rates of fish were considerably increased from 10.0% in the control group to 60.0% and 66.7% in the BSFO80 and BSFO100 groups, respectively. One-variable linear regression analysis revealed that various parameters related to fish growth, flesh quality, and health status were significantly influenced by dietary BSFO substitution levels in a dose-dependent manner (P < 0.05). In conclusion, substituting around 20% of dietary fish oil and soybean oil with BSFO is promising in improving the growth performance and flesh quality of M. salmoides. However, to enhance immunity and disease resistance, it is recommended to further increase the inclusion of BSFO in the diet.

Giant intracranial arachnoid cyst in an outstanding soldier.

This article reports a case of a large intracranial arachnoid cyst in a soldier who had performed well. This is a very marvelous and interesting case. Although the cyst pressed the entire brain into the contralateral cranial cavity, the patient did not show any significant clinical symptoms and had excellent motor and cognitive abilities, reflecting the excellent plasticity of the brain tissue.

Effects of Different Defatting Methods of Black Soldier Fly (Hermetia illucens) Larvae Meal on the Metabolic Energy and Nutrient Digestibility in Young Laying Hens.

This study aimed to investigate the effects of different defatting methods of black soldier fly (Hermetia illucens) larvae meal (BSFM) on the metabolic energy and nutrient digestibility in laying hens. Sixty young laying hens (Hy-Line W-36) aged 63 days were randomly divided into two groups (G1 and G2), each with five replicates of six hens housed in individual cages. Group G1 was fed 25% pressed black soldier fly meal (BSFMp) and 75% basal diet, and Group G2 was fed 25% extracted black soldier fly meal (BSFMe) and a 75% basal diet. Both diets included 5 g/kg chromium oxide as an external marker. A 7-day preliminary trial was followed by a 4-day experimental period. The results indicate that pressing and extracting significantly affected the digestibility of crude fat and total energy in BSFM, with BSFMp showing significantly higher crude fat digestibility than BSFMe. Similarly, total energy digestibility was also significantly higher in BSFMp. However, there were no significant differences in dry matter, organic matter, and crude protein digestibility between the two processing methods. The apparent metabolic energy values of BSFMp and BSFMe were 16.34 and 12.41 MJ/kg, respectively, showing a significant difference. The nitrogen-corrected metabolic energy values were 15.89 MJ/kg in BSFMp and 11.93 MJ/kg in BSFMe, indicating a highly significant difference. The digestibility of arginine and leucine in BSFMp was significantly higher than in BSFMe, while differences in lysine, cystine, threonine, tryptophan, and isoleucine were not significant. In conclusion, both defatting methods of BSFM had no adverse effects on the metabolic energy and nutrient digestibility in young laying hens, but BSFMp demonstrated better effects on the digestibility of metabolic energy and nutrients in the feed for young laying hens.

The Influence of Alternative Diets and Whole Dry Black Soldier Fly Larvae (Hermetia illucens) on the Production Performance, Blood Status, and Egg Quality of Laying Hens.

Given the significant environmental consequences of current poultry feed practices and the heavy dependence of the European Union on imported soybeans, studying alternatives is crucial. This study evaluated the potential benefits of using locally sourced alternative plant-based ingredients and whole dry black soldier fly larvae in the diet of laying hens. The experiment involved 120 Isazul hens at 23 weeks old, which were divided into three groups with five replicates each (eight hens per replicate): a control diet (CON) based on soybean meal and cereals, an alternative diet (ALT) replacing the soybean meal with locally sourced plant-based resources (peas, distillers' dried grains with solubles, and sunflower meal), and the ALT diet supplemented with 5% whole dry black soldier fly larvae (ALT+DBSFL). Over 15 weeks, the hens were fed ad libitum, and the production performance, egg quality, and plasma biochemical parameters were assessed across three experimental sub-periods. The results showed no significant differences in body weight, feed intake, egg production, egg weight, egg mass, or feed conversion ratio across the diets (p > 0.05). The egg quality remained consistent across all the groups; however, the hens fed the ALT+DBSFL diet exhibited higher Haugh units in the first experimental sub-period (p < 0.05) and lower plasma cholesterol and triglycerides at 32 weeks of age (p < 0.05). The findings of this study indicate that incorporating these alternative ingredients and whole DBSFL into layers' diets does not compromise production performance, egg quality, or biochemical parameters related to health status, supporting their potential as sustainable feed alternatives.

Willingness of West African Consumers to Buy Food Produced Using Black Soldier Fly Larvae and Frass.

The use of black soldier fly (BSF) larvae and frass in agriculture can make an important contribution to food and nutrition security. However, it is important to understand whether consumers are willing to consume food products resulting from the use of BSF larvae as animal feed or BSF frass as fertilizer. This study employed the stated preference approach as food products produced using BSF larvae and frass are not currently available on the market. Questionnaires were administered to a total of 4412 consumers in Ghana (1360), Mali (1603), and Niger (1449). The results show that the vast majority of respondents are willing to consume vegetables (88%) produced using BSF frass and meat (87%) produced using animal feed made of BSF larvae. A smaller percentage of respondents are even willing to pay USD 1.32 and USD 1.7 more if the base price of BSF-based products were USD 5 per kg. Age, gender, education, and country positively influenced the respondents' willingness to consume food produced using BSF products. In contrast, neighborhood status, income, and household size are inversely related to the respondents' willingness to pay for and consume these products. Our findings are, therefore, important to scaling up BSF technologies in the region.

Hormesis of black soldier fly larva: Influence and interactions in livestock manure recycling.

Black soldier fly larvae (BSFL) are considered important organisms, utilized as tools to transform waste including manure into valuable products. The growth and cultivation of BSFL are influenced by various factors, such as the presence of toxic substances in the feed and parasites. These factors play a crucial role in hormesis, and contributing to regulate these contaminants hermetic doses to get sustainable byproducts. This review aims to understand the effects on BSFL growth and activities in the presence of compounds like organic and inorganic pollutants. It also assesses the impact of microbes on BSFL growth and explores the bioaccumulation of pharmaceutical compounds, specifically focusing on heavy metals, pesticides, pharmaceuticals, indigenous bacteria, insects, and nematodes. The review concludes by addressing knowledge gaps, proposing future biorefineries, and offering recommendations for further research.

Effect of black soldier fly larvae frass addition on humus content during low temperature co-composting.

Initiating aerobic fermentation under low temperature is the main challenge for winter livestock manure composting. This study aims to address this issue by applying black soldier fly larvae (BSFL) frass as a co-composting additive to enhance the low-temperature composting process. Specifically, this work explored the effects of chicken manure and BSFL frass co-composting on the temperature, humus content, and microorganisms with fresh weight ratio of 2:1, 1:1, 1:2 (w/w) at 6 °C. The result showed frass could rapidly rise the temperature to 50 °C and significantly increased the humus content by 15.6 % ∼ 26.3 %. Moreover, microbial analysis revealed that Sphingobacteriaceae accelerated temperature rise via low-temperature reproduction, creating proper temperature for thermophilic bacteria (Truepera and Georgia). Additionally, Cellulomonas and other bacteria promoted organic matter degradation and participated in humus formation. This study presents a novel solution for low-temperature composting, providing practical insights for improving manure management in winter.