Search Results (69)

This study aims to investigate the effects of selenium (Se) fortification on growth performance and the Se content in kale using Se fertilizer, and it determines the influences of Se fortification on the metabolic profile of kale using quasi-targeted metabolomics. The results showed that Se fortification increased the plant height and leaf weight of kale, up-regulated the total Se content and decreased the chlorophyll and total phenolic contents in kale leaf. Se fortification elevated selenate (Se(IV)), selenite (Se(VI)), selenocystine (SeCys₂), Se-methylselenocysteine (Se-MeSeCys) and selenomethionine (SeMet) contents, as well as total contents of Se in different forms in kale leaf. Se fortification also changed the metabolic profile of kale leaf, via six particular types of compounds (amino acid and its derivatives; organic acid and its derivatives; carbohydrates and its derivatives; lipids; flavonoids; organoheterocyclic compounds) and eight metabolic pathways (alanine, aspartate and glutamate metabolism; amino sugar and nucleotide sugar metabolism; sulfur metabolism; starch and sucrose metabolism; taurine and hypotaurine metabolism; glycolysis/gluconeogenesis; fructose and mannose metabolism; nitrogen metabolism). Moreover, 24 metabolic biomarkers were screened for kale leaf affected by Se fortification. Furthermore, correlations were observed between metabolic biomarkers and Se contents as well as speciation. These results indicate that Se fortification has a significant influence on the growth performance and nutritional compounds of kale, providing references for the future study on the production and bioactivity of Se-enriched kale. Full article

The research aimed to examine the effects of fermentation and enzymatic hydrolysis of cottonseed protein on body weight changes, serum biochemistry, rumen function, intestinal health, and liver metabolism of suckling lambs. A total of twelve 7-day-old healthy male Hu sheep body weights (5.27 ± 0.48 kg) were randomly distributed into two groups. Starter feed regimens containing microbial fermentation of cottonseed protein (MFCP) or enzymatic hydrolysate of cottonseed protein (EHCP) were administered to lambs during the initial 60-day period. Results showed that compared with EHCP group, the serum glucose, ruminal acetic, propionic, butyric and valeric acids concentrations, jejunal immunoglobulin G content and mRNA expressions of Claudin 1 and Occludin, as well as the relative abundance of actinobacteriota and pseudoscardovia in the rumen were significantly increased in the MFCP group (p < 0.05), whereas an opposite trend was observed in the jejunum. α-amylase and trypsin enzymatic activities were observed between the two groups. Relative to EHCP treatment, the MFCP group exhibited 69 elevated and 103 reduced hepatic metabolites, and these metabolites displayed distinct enrichment patterns within specific metabolic networks, including fructose and mannose metabolism (p = 0.003), arachidonic acid metabolism (p = 0.017), glycerophospholipid metabolism (p = 0.036), and the cAMP signaling pathway (p = 0.047). Overall, microbial fermentation of cottonseed protein may be beneficial for strengthening intestinal barrier function and facilitating hepatic lipid metabolism and immune regulation, while enzymatic hydrolysis of cottonseed protein enhances gastrointestinal digestive enzyme activity, thereby promoting nutrient digestion of suckling lambs. Full article

Background: The dysregulation of both glucose and lipid metabolism is the main clinical features of type 2 diabetes. Qihua Tongtiao Formula (QHTTF) is our team’s current clinical empirical formula, and the related patent has been granted. It is composed of Astragalus membranaceus, Atractylodes macrocephala koidz, Aurantii Fructus Immaturus, Radix Bupleuri, Ligusticum chuanxiong hort, Angelicae sinensis radix, Raphanus sativus, and Polyporus umbellatus. It can alleviate tissue pathological damage in type 2 diabetic rats by improving glycolipid metabolism disorders. Nevertheless, the specific mechanisms of QHTTF in the treatment of type 2 diabetes remain unclear. Purpose: This research aims to explore the fundamental effect and underlying mechanism of the QHTTF formula in ZDF rats via network pharmacology, biological validation, and metabolomics technology. Methods: The chemical compounds of QHTTF were initially identified via UHPLC-MS/MS analysis. Meanwhile, drug targets, genes, related diseases, and differential metabolites of QHTTF in the treatment of T2DM were obtained through network pharmacology, molecular docking, and metabolomics. Then, we conducted animal experiments to further explore the therapeutic molecular mechanism of QHTTF in ZDF rats. Results: A total of 39 main chemical components were recognized through LC-MS/MS technology, and 22 remained after the second screening. Network pharmacology and molecular docking results revealed that 59 intersection targets were involved in the treatment of glycolipid metabolic disorders, and the PPARα, PPARγ, and TNF proteins were identified as crucial targets through PPI network analysis. Additionally, serum metabolomics analysis of ZDF rats showed that QHTTF could regulate linoleic acid metabolism, fructose and mannose metabolism, galactose metabolism, fatty acid biosynthesis, and other related signaling pathways. The results of biological experiments proved that QHTTF effectively lowered blood glucose and lipid levels, alleviated hepatic and pancreatic pathological damage, increased the expression of IRS-1 and GLUT4 in the pancreas, and improved insulin resistance, while inhibiting the inflammatory response and oxidative stress, as well as enhancing the expression of liver PPARα, PPARγ, and AMPK proteins in ZDF rats. Conclusions: In summary, QHTTF exerted a significant effect in improving glycolipid metabolism disorders of ZDF rats, which might show therapeutic effects by relieving insulin resistance, mitigating inflammation and oxidative damage, regulating related glucose, fatty acid, and amino acid metabolism, and increasing the expression of PPARα, PPARγ, and AMPK proteins by combining network analysis, metabolomics, and biological research. Full article

Xylazine abuse is emerging as a global problem, whereas the toxic mechanisms of xylazine poisoning are seldom studied. The present study aims to assess the heart injury in xylazine poisoning and uncover the underlying mechanism. Forty male SD rats were randomly dived into four groups: control (saline), low dose (10 mg/kg xylazine), median dose (20 mg/kg xylazine) and high dose (40 mg/kg xylazine). The rats were injected with the drug intraperitoneally for 28 consecutive days, and then cardiac ultrasound examination was performed and serum and heart tissues were collected. Genomic, proteomic, and metabolic omics analyses were conducted. ELISA, RNA sequencing, histopathology examination, RT-qPCR, and Western blot were performed. Repeated injection of xylazine led to a decrease in the expression of cardiac output (CO), ventricular systole (VS), and ventricular diastole (VD), while concurrently elevating the levels of lactate dehydrogenase (LDH), creatine kinase myocardial band (CK-MB), and cardiac troponin T (c-TNT) in the serum. HE staining analysis showed evidence of contraction band necrosis, interstitial fibrosis, and infiltration by inflammatory cells in animals with xylazine poisoning. The modified genes, proteins, and metabolites were gathered, and the integration of transcriptomic, proteomic, and metabolic networks identified 25 overlapping pathways between the differentially expressed genes and metabolites (DEGs-DEMs) and the differentially expressed proteins and metabolites (DEPs-DEMs) joint pathways. The majority of these pathways pertained to the metabolism of sugars, amino acids, and fats. The proteins associated with fructose and mannose metabolism, as well as cholesterol metabolism, were validated, thereby substantiating their pivotal role in the development of xylazine-induced cardiac injury. Repeated injection of xylazine impaired heart function and the metabolism of fructose and mannose. Cholesterol metabolism pathways were critical in the process of xylazine-induced heart injury. Full article

To investigate the impacts of dietary Bacillus-based probiotics and yeast-derived prebiotics on the hepatic transcriptome profile, 500 Hisex White laying hens were randomly allotted into five dietary treatments from 37 to 52 weeks of age: control; control + Bacillus subtilis; control + Bacillus subtilis and Bacillus licheniformis; control + Bacillus coagulans; and control + Saccharomyces cerevisiae yeast cell wall. Transcriptome analysis revealed a substantial number of differentially expressed genes exclusively between the control and prebiotic groups, identifying 2221 genes (FDR ≤ 0.05), with 980 genes upregulated (log₂ fold change 0.69 to 24.62) and 1241 downregulated (log₂ fold change −0.74 to −26.46). The top 10 upregulated KEGG pathways included protein export, glycerophospholipid metabolism, tryptophan metabolism, amino acid biosynthesis, alanine, aspartate, and glutamate metabolism, cofactor biosynthesis, propanoate metabolism, ABC transporters, 2-oxocarboxylic acid metabolism, and protein processing within the endoplasmic reticulum. In contrast, the most prominently downregulated pathways encompassed fructose and mannose metabolism, hedgehog signaling, PPAR signaling, Notch signaling, GnRH signaling, cell adhesion molecules, cytokine–cytokine receptor interactions, apelin signaling, glycosaminoglycan degradation, and RIG-I-like receptor signaling. These findings advance understanding of the hepatic transcriptomic response to yeast-derived prebiotics and identify key molecular pathways that could be targeted to enhance metabolic function in laying hens. Full article

Background/Objectives: Blunt cardiac injury (BCI) is a severe medical condition that may arise as a result of various traumas, including motor vehicle accidents and falls. The main objective of this study was to explore the role and underlying mechanisms of the TRPV4 gene in BCI. Elucidating the function of TRPV4 in BCI may reveal potential novel therapeutic targets for the treatment of this condition. Methods: Rats in each group, including the SD control group (SDCON), the SD blunt-trauma group (SDBT), the TRPV4 gene-knockout control group (KOCON), and the TRPV4 gene-knockout blunt-trauma group (KOBT), were all freely dropped from a fixed height with a weight of 200 g and struck in the left chest with a certain energy, causing BCI. After the experiment, the levels of serum IL-6 and IL-1β were detected to evaluate the inflammatory response. The myocardial tissue structure was observed by HE staining. In addition, cardiac transcriptome analysis was conducted to identify differentially expressed genes, and metabolomics studies were carried out using UHPLC-Q-TOF/MS technology to analyze metabolites. The results of transcriptomics and metabolomics were verified by qRT-PCR and Western blot analysis. Results: Compared with the SDCON group, the levels of serum IL-6 and IL-1β in the SDBT group were significantly increased (p < 0.001), while the levels of serum IL-6 and IL-1β in the KOBT group were significantly decreased (p < 0.001), indicating that the deletion of the TRPV4 gene alleviated the inflammation induced by BCI. HE staining showed that myocardial tissue injury was severe in the SDBT group, while myocardial tissue structure abnormalities were mild in the KOBT group. Transcriptome analysis revealed that there were 1045 upregulated genes and 643 downregulated genes in the KOBT group. These genes were enriched in pathways related to inflammation, apoptosis, and tissue repair, such as p53, apoptosis, AMPK, PPAR, and other signaling pathways. Metabolomics studies have found that TRPV4 regulates nucleotide metabolism, amino-acid metabolism, biotin metabolism, arginine and proline metabolism, pentose phosphate pathway, fructose and mannose metabolism, etc., in myocardial tissue. The combined analysis of metabolic and transcriptional data reveals that tryptophan metabolism and the protein digestion and absorption pathway may be the key mechanisms. The qRT-PCR results corroborated the expression of key genes identified in the transcriptome sequencing, while Western blot analysis validated the protein expression levels of pivotal regulators within the p53 and AMPK signaling pathways. Conclusions: Overall, the deletion of the TRPV4 gene effectively alleviates cardiac injury by reducing inflammation and tissue damage. These findings suggest that TRPV4 may become a new therapeutic target for BCI, providing new insights for future therapeutic strategies. Full article

Insights into dynamic regulatory factors in various stages of growth and development can guide strategies for precision and targeted breeding. Bitter gourd, as a vegetable product with medicinal value, plays a role in both agricultural and medical fields. In this study, phenotypic observations, metabolomic and transcriptomic analyses, and differential gene expression patterns, along with a correlation analysis, were conducted in different stages of fruit growth and development. The results revealed that the growth rate of fruit’s fresh weight, length, diameter, and flesh thickness during the first seven days was slow, and that it then rapidly increased after the seventh day, and finally slowed once more after 17 days, indicating that the overall process followed a “slow–fast–slow” pattern. Transcriptomic and metabolomic analyses identified several differentially expressed genes and metabolites, and joint analyses revealed that each of the glycolysis/gluconeogenesis, fructose and mannose metabolism and flavonoid biosynthesis pathways individually play significant roles in the dynamic regulation of fruit growth and development during the early, middle, and late stages. Among these, 53 differentially expressed genes (DEGs) and 12 differentially expressed metabolites (DEMs) were found in these pathways. A total of 12 randomly selected DEGs were analyzed using quantitative PCR, and the results showed that gene expression levels were generally consistent with transcriptomic sequencing results, exhibiting dynamic changes with varying expression levels. Correlation analysis revealed that 11 DEMs were positively correlated with four traits except for arbutin, while eight DEGs were related to all traits, including six significantly positive and two significantly negative correlations. These findings enhance our understanding of the regulatory network governing yield and quality and provide substantial evidence to support improvements in breeding programs. Full article

Low temperature is a major abiotic stress affecting rice productivity. Selenium (Se) treatment has been shown to enhance plant resilience to cold stress. In this study, low concentrations of selenium (ColdSe1) alleviated the adverse effects of cold stress on rice seedlings, improving fresh weight, plant height, and chlorophyll content by 36.9%, 24.3%, and 8.4%, respectively, while reducing malondialdehyde (MDA) content by 29.1%. Se treatment also increased the activities of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), by 25.2%, 42.7%, and 33.3%, respectively, and upregulated flavonoids, soluble sugars, cysteine (Cys), glutathione (GSH), and oxidized glutathione (GSSG). Transcriptome analysis revealed that ColdSe1 treatment upregulated genes associated with amino and nucleotide sugar metabolism, glutathione metabolism, and fructose and mannose metabolism. It also alleviated cold stress by modulating the MAPK signaling pathway, phytohormone signaling, and photosynthesis-related pathways, enriching genes and transcription factors linked to antioxidant metabolism and photosynthesis. Metabolomic analyses showed that ColdSe1 positively influenced amino acid glucose metabolism, glycerolipid metabolism, hormonal pathways, and alanine/glutamate pathways under cold stress, while also upregulating metabolites associated with plant secondary metabolites (e.g., flavonoids, phenolic compounds) and antioxidant metabolism (e.g., α-linolenic acid metabolism). In contrast, high selenium concentrations (ColdSe2) disrupted phenylpropanoid biosynthesis, α-linolenic acid metabolism, and ABC transporter function, exacerbating cold-stress injury. This study highlights the critical role of Se in mitigating cold stress in rice, offering a theoretical basis for its application as an agricultural stress reliever. Full article

Hydrogen peroxide (H₂O₂) functions as a signaling molecule that triggers physiological and biochemical adjustments that help plants cope with environmental stress. This study evaluated the effects of foliar application of 1.5 mM H₂O₂ on the physiological and biochemical responses of Passiflora incarnata subjected to 14 days of drought stress followed by 5 days of rehydration. Drought reduced Fv/Fm and photochemical efficiency, as well as stomatal conductance and transpiration rates. H₂O₂ treatment under drought further reduced stomatal conductance and transpiration, suggesting enhanced water conservation. Drought-stressed plants treated with H₂O₂ exhibited increased concentrations of glucose, fructose, and mannose along with reduced sucrose levels, indicating osmotic adjustment and energy mobilization. Enzymatic antioxidant activity, particularly that of superoxide dismutase and catalase, increased with H₂O₂ treatment, while peroxidase activity remained low. The content of vitexin, arabinose, and trehalose decreased under drought, likely due to their roles in membrane protection, as MDA levels remained stable. After rehydration, Fv/Fm and ΦPSII recovered, and H₂O₂-treated plants showed higher carbon assimilation and carboxylation efficiency. These results indicate that H₂O₂ promotes drought acclimation and enhances post-stress recovery in P. incarnata. We conclude that H₂O₂ induces signaling pathways, with trehalose, arabinose, and vitexin contributing to the regeneration of the photochemical apparatus, as well as defense and acclimation under drought conditions. Full article

Drought stress is an important abiotic stress factor restricting crop production. Broomcorn millet (Panicum miliaceum L.) has become an ideal material for analyzing the stress adaptation mechanisms of crops due to its strong stress resistance. However, the functional characteristics of its rhizosphere microorganisms in response to drought remain unclear. In this study, metagenomics and metabolomics techniques were employed to systematically analyze the compositional characteristics of the microbial community, functional properties, and changes in metabolites in the rhizosphere soil of broomcorn millet under drought stress. On this basis, an analysis was conducted in combination with the differences in functional pathways. The results showed that the drought treatment during the flowering stage significantly altered the species composition of the rhizosphere microorganisms of broomcorn millet. Among them, the relative abundances of beneficial microorganisms such as Nitrosospira, Coniochaeta, Diversispora, Gigaspora, Glomus, and Rhizophagus increased significantly. Drought stress significantly affects the metabolic pathways of rhizosphere microorganisms. The relative abundances of genes associated with prokaryotes, glycolysis/gluconeogenesis, and other metabolic process (e.g., ribosome biosynthesis, amino sugar and nucleotide sugar metabolism, and fructose and mannose metabolism) increased significantly. Additionally, the expression levels of functional genes involved in the phosphorus cycle were markedly upregulated. Drought stress also significantly alters the content of specific rhizosphere soil metabolites (e.g., trehalose, proline). Under drought conditions, broomcorn millet may stabilize the rhizosphere microbial community by inducing its restructuring and recruiting beneficial fungal groups. These community-level changes can enhance element cycling efficiency, optimize symbiotic interactions between broomcorn millet and rhizosphere microorganisms, and ultimately improve the crop’s drought adaptability. Furthermore, the soil metabolome (e.g., trehalose and proline) functions as a pivotal interfacial mediator, orchestrating the interaction network between broomcorn millet and rhizosphere microorganisms, thereby enhancing plant stress tolerance. This study sheds new light on the functional traits of rhizosphere microbiota under drought stress and their mechanistic interactions with host plants. Full article

High temperatures in summer often trigger disease outbreaks in shrimp, resulting in significant economic losses. To investigate the heat tolerance mechanisms of Penaeus monodon, juvenile tiger shrimp were subjected to a high-temperature stress of 38 °C for 144 h. The cumulative survival rate of shrimp sharply decreased to 5.29% in the later 144 h. The heat-sensitive shrimps (S group) were collected in the first 24 h, while those that survived beyond 120 h were collected as the heat-tolerant group (T group). The hepatopancreas of two groups was subjected to transcriptomic and metabolomic analysis. The results revealed that, compared to the S group, the T group exhibited a total of 3527 DEGs, including 2199 upregulated and 1328 downregulated genes. Additionally, 353 DAMs were identified in the T group, with 75 metabolites showing increased levels and 278 metabolites displaying decreased levels. The results revealed that the mechanisms of heat tolerance involve energy supply strategies, immune system regulation, amino acid metabolism, and glutathione metabolism. Energy supply strategies include the digestion and absorption of carbohydrates and proteins, glycolysis/gluconeogenesis, fructose and mannose metabolism, and pyruvate metabolism, all of which collectively meet energy demands in high-temperature environments. The immune system is regulated by C-type lectin receptor pathways and IL-17 signaling pathways, which together coordinate innate immunity to prevent pathogen invasion. In amino acid metabolism, various glycogenic amino acids, such as histidine, phenylalanine, valine, and serine, are metabolized for energy, while excess ammonia is converted to γ-glutamyl-glutamate and L-glutamate to mitigate ammonia accumulation. Combined transcriptomic and metabolomic analyses further indicate that glutathione metabolism plays a crucial role in the adaptation of P. monodon to high-temperature environments. This study explains the high-temperature tolerance mechanism of P. monodon from the aspects of gene expression regulation and material metabolism regulation and also provides a scientific basis and basic data for the selection and breeding of new varieties of P. monodon with a high-temperature tolerance. Full article

The ladybird beetle, Propylea japonica Thunberg (Coleoptera: Coccinellidae), is a widely distributed natural predator that is crucial in controlling various agricultural pests in China. Despite frequent references to its remarkable thermotolerance, the molecular mechanisms underlying its thermotolerance remain poorly understood. Here, we investigated metabolomic changes in P. japonica following exposure to acute heat stress (AHS) lasting 1 h at 39 °C and 43 °C in populations from Zhengzhou (ZZ, warm temperate climate zone) and Shenzhen (SZ, subtropical climate zone), representing distinct northern and southern Chinese ecosystems. A total of 4165 and 4151 metabolites were detected in positive and negative ion modes, respectively. The high proportion of lipid and lipid-like metabolites (35.5%) and the top 20 pathways containing the highest number of metabolites, implying membrane fluidity modulation and energy metabolism restructuring, served as the core adaptive mechanism in P. japonica populations confronting thermal stress. The SZ25 vs. SZ39 exhibited a significantly higher number of differentially expressed metabolites (DEMs), which were predominantly enriched in the purine and tryptophan metabolism pathways. This indicated that these pathways orchestrate thermal adaptation in the SZ population by coordinating energy metabolism reprogramming, orchestrating antioxidant defense mechanisms, and modulating neuroendocrine homeostasis dysregulation. Additionally, the starch and sucrose, arachidonic acid, and fructose and mannose metabolism pathways were also implicated. This study enhances our understanding of P. japonica thermotolerance and provides a valuable reference for thermotolerance mechanisms in other insects. Full article

Perfluoroctane sulfonate (PFOS) is an emerging pollutant widely existing in aquatic environments that has attracted many scholars’ attention. Cherax quadricarinatus (C. quadricarinatus) are crustaceans that live in freshwater environments. This study aimed to investigate the long-term toxic exposure effect of PFOS on C. quadricarinatus. Three PFOS environment concentrations (1 ng/L, 100 ng/L, and 10 μg/L) were set for 28 days of exposure to C. quadricarinatus. The results indicated that PFOS was detected in the serum, muscle, and hepatopancreas of the C. quadricarinatus, and the order of accumulation levels was as follows: hepatopancreas > serum > muscle. Furthermore, transcriptomics showed that the function of differentially expressed genes (DEGs) in PFOS exposure groups was related to biological processes, metabolism, organic system, and immune response. The Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that DEGs were significantly enriched in the lysosome signaling pathway, retinol binding, fructose and mannose metabolism, and glutathione metabolism, etc., and the lysosome signaling pathway was the most significant, which indicated that lysosome signaling pathway is the key pathway for the toxic effects of PFOS on C. quadricarinatus. Full article

Background/Objectives: The largemouth bass (Micropterus salmonides) is a farmed fish of significant economic value, and studying its adaptability is crucial for enhancing the economic benefits of aquaculture. The largemouth bass changes gene expression pattern to rapidly adapt to environmental changes and maintain normal physiological function. Methods: In this study, largemouth bass from two distinct environmental backgrounds—Huzhou and Xingtai—were used as experimental materials, and they have significantly different breeding conditions. Comparative transcriptomics was used to analyze the gene expression patterns in largemouth bass from both backgrounds. Results: In the female, there were 1678 differentially expressed genes, of which 541 were upregulated and 1137 were downregulated. Meanwhile, in the male, there were 1287 differentially expressed genes, including 542 upregulated genes and 745 downregulated genes. The differentially expressed genes were mainly enriched in biological processes such as metabolic process, biological regulation, response to stimulus, developmental process, signaling, reproduction and immune system process. The enriched pathways included carbon metabolism, glycolysis/gluconeogenesis, purine metabolism, biosynthesis of amino acids, starch and sucrose metabolism, fructose and mannose metabolism, pyrimidine metabolism, MAPK signaling pathway, spliceosome, protein processing in the endoplasmic reticulum, ribosome biogenesis in eukaryotes, etc. Conclusions: We speculated that largemouth bass in Xingtai may adapt to the environment by downregulating metabolism- and reproduction-related genes and altering the expression of immune-related genes. Our study provided molecular evidence for the adaptation research of largemouth bass and provided a scientific basis for optimizing largemouth bass breeding technology. Full article

Background/Objectives: miRNAs are a family of single-stranded non-coding RNAs that regulate gene expression by targeting messenger RNAs (mRNAs) for suppression, with an average length of 22 nt. The oriental armyworm, Mythimna separata Walker, is a pest insect with long-distance migratory capability, which causes severe loss of grains and pastures in Eastern Asia, Southeastern Asia, and Oceania. This study aims to elucidate the post-transcriptional regulatory mechanisms of miRNAs in the development of this pest. Methods: We carried out small RNA sequencing on samples from eggs, third instar larvae, pre-pupae, pupae, and adults. Results: A total of 400 miRNAs were identified, among which 40 were known and 360 were novel miRNAs. Dynamic trend analysis of miRNAs revealed that 199 miRNAs were highly expressed in eggs (profile 12), while 173 miRNAs were highly expressed in both eggs and pupae (profile 13). The results of differential expression analysis of miRNAs (DEmiR) revealed that 75 miRNAs were significantly more abundant in eggs compared to other developmental stages. Furthermore, more up-regulated miRNAs were observed than down-regulated miRNAs in adults relative to 3rd instar larvae, pre-pupae, and pupae. The core genes for miRNA biosynthesis—Pasha, Dicer1, and Ago1—were highly expressed in eggs but poorly expressed in 3rd instar larvae. KEGG enrichment analyses indicated that several genes in the pentose and glucuronate interconversion pathway, as well as the fructose and mannose metabolism pathway, were regulated by DEmiRs. Conclusions: DEmiRNAs targeted most genes of M. separata, resulting in a complex miRNA–mRNA regulation mode. Full article