Paper-based fluorescence sensor array with functionalized carbon quantum dots for bacterial discrimination using a machine learning algorithm.

The rapid discrimination of bacteria is currently an emerging trend in the fields of food safety, medical detection, and environmental observation. Traditional methods often require lengthy culturing processes, specialized analytical equipment, and bacterial recognition receptors. In response to this need, we have developed a paper-based fluorescence sensor array platform for identifying different bacteria. The sensor array is based on three unique carbon quantum dots (CQDs) as sensing units, each modified with a different antibiotic (polymyxin B, ampicillin, and gentamicin). These antibiotic-modified CQDs can aggregate on the bacterial surface, triggering aggregation-induced fluorescence quenching. The sensor array exhibits varying fluorescent responses to different bacterial species. To achieve low-cost and portable detection, CQDs were formulated into fluorescent ink and used with an inkjet printer to manufacture paper-based sensor arrays. A smartphone was used to collect the responses generated by the bacteria and platform. Diverse machine learning algorithms were utilized to discriminate bacterial types. Our findings showcase the platform's remarkable capability to differentiate among five bacterial strains, within a detection range spanning from 1.0 × 103 CFU/mL to 1.0 × 107 CFU/mL. Its practicality is further validated through the accurate identification of blind bacterial samples. With its cost-effectiveness, ease of fabrication, and high degree of integration, this platform holds significant promise for on-site detection of diverse bacteria.

Metformin promotes bacterial surface aggregation by inhibiting the swimming motility of flagellated Escherichia coli.

The antiglycemic drug metformin, which is widely prescribed as a first-line drug for the treatment of type 2 diabetes, has become a concern for emerging pollutants in natural ecosystems. However, its effects on bacterial swimming motility remain unclear. In this study, we showed that metformin promotes bacterial surface aggregation by tracking swimming and by measuring the density distribution of Escherichia coli cultured with metformin near a surface in a homogeneous environment. Flagella are essential for the promotion of bacterial surface aggregation by metformin. Swimming motility, which is mediated by flagella, determines bacterial surface aggregation. The promotion of bacterial surface aggregation by metformin is caused by a reduction in swimming motility, which is governed by a decrease in the proton motive force. Our results reveal that metformin has a pronounced effect on flagellated bacterial motility associated with surface sensing and aggregation.

Diet Supplementation of Luteolin before Fatty Liver Formation Improves Hepatic Steatosis in Obese Mice by Inhibiting Visceral Adipose Tissue Lipolysis.

SCOPE: Serotonin (5-HT)-induced visceral adipocyte lipolysis is essential for the development of obesity-related complications. Diet supplementation of luteolin prevents high-fat diet (HFD)-fed mice against obesity and associated fatty liver. However, independent of the body weight loss, whether dietary luteolin can substantially reduce hepatic steatosis remains unclear. METHODS AND RESULTS: In differentiated 3T3-L1 cells, 5-HT treatment promotes adipocyte lipolysis, while luteolin significantly inhibits 5-HT-induced lipolysis, Ca2+ -PKG cascade, and SIRT1/FoxO1/AMPKα signaling through binding to 5-HT receptor HTR2B. Further, 5-week-old mice are fed with an HFD for 16 weeks. At the 6th, 8th, or 10th weeks of HFD feeding, some mice are switched to a luteolin-containing HFD, respectively. In all HFD-fed mice, body weight gain and body component are unaffected by dietary luteolin. However, diet supplementation of luteolin at the 6th or 8th, rather than at the 10th weeks, alleviates hepatic steatosis. Meanwhile, dietary luteolin reduces epididymal adipose tissue (EAT) lipolysis, and represses the level of lipolytic enzyme, the expression of Htr2b, and the activation of PKG and SIRT1/FoxO1/AMPKα signaling in EAT. CONCLUSIONS: Diet supplementation of luteolin before the formation of fatty liver protects HFD-fed mice against ectopic lipid deposition in liver by inhibiting visceral adipocyte lipolysis.

The activity comparison of six dietary flavonoids identifies that luteolin inhibits 3T3-L1 adipocyte differentiation through reducing ROS generation.

Mitochondrial reactive oxygen species (ROS)generation plays an essential role in the process of adipocyte differentiation and is involved in the development of obesity and associated metabolic diseases. Various dietary flavonoids possess the substantial anti-adipogenic activity. However, it is unclear whether these flavonoids inhibit adipocyte differentiation by reducing ROS generation. In this study, the effects of six common dietary flavonoids on adipocyte differentiation were assessed in 3T3-L1 cells. The flavonoids with the same backbone of 5,7-dihydroxylflavone, including flavones apigenin, chrysin, luteolin and flavonols kaempferol, myricetin, quercetin, dose-dependently inhibited 3T3-L1 adipocyte differentiation, suggesting an associated hierarchy of inhibitory capability: luteolin > quercetin > myricetin > apigenin/kaempferol > chrysin. Meanwhile, six flavonoids were found to inhibit adipogenic gene expression and the early stage of adipocyte differentiation. Among the tested flavonoids, luteolin significantly reduced both intracellular and mitochondrial ROS generation during adipocyte differentiation. Further, luteolin treatment depressed the elevation of H2O2 concentration in the early stage of 3T3-L1 differentiation and reversed the facilitated effects of exogenous H2O2 on 3T3-L1 adipocyte differentiation and ROS generation. Altogether, the activity comparison of six dietary flavonoids identifies that luteolin inhibits 3T3-L1 adipocyte differentiation through reducing ROS generation, elucidating a new mechanism underlying the anti-adipogenic actions of flavonoids.

Lysosome Inhibition Reduces Basal and Nutrient-Induced Fat Accumulation in Caenorhabditis elegans.

A long-term energy nutritional imbalance fundamentally causes the development of obesity and associated fat accumulation. Lysosomes, as nutrient-sensing and lipophagy centers, critically control cellular lipid catabolism in response to nutrient deprivation. However, whether lysosome activity is directly involved in nutrient-induced fat accumulation remains unclear. In this study, worm fat accumulation was induced by 1 mM glucose or 0.02 mM palmitic acid supplementation. Along with the elevation of fat accumulation, lysosomal number and acidification were also increased, suggesting that lysosome activity might be correlated with nutrient-induced fat deposition in Caenorhabditis elegans. Furthermore, treatments with the lysosomal inhibitors chloroquine and leupeptin significantly reduced basal and nutrient-induced fat accumulation in C. elegans. The knockdown of hlh-30, which is a critical gene in lysosomal biogenesis, also resulted in worm fat loss. Finally, the mutation of aak-2, daf-15, and rsks-1 showed that mTORC1 (mechanistic target of rapamycin complex-1) signaling mediated the effects of lysosomes on basal and nutrient-induced fat accumulation in C. elegans. Overall, this study reveals the previously undescribed role of lysosomes in overnutrition sensing, suggesting a new strategy for controlling body fat accumulation.

Characterization of the response of Escherichia coli to l-fucose in bacterial swimming motility.

l-Fucose, as a monosaccharide in nature, plays a crucial role in bacteria colonization. Escherichia coli (E. coli), as a common microorganism in environment, utilize bacterial flagellar motor to drive the rotation of flagella, which is regulated by chemotactic signal transduction signals. Yet the effect of l-fucose to bacterial motility remains unclear. The effect of l-fucose on the swimming motility of bacteria was investigated from the level of single flagellar motor to individual cell and cell population by employing a bead assay, a high-throughput 2D tracking assay and a high-throughput dark-field flicker microscopy. The results showed that the swimming motility of the bacteria cultured with l-fucose was decreased, while the tumble frequency increased. Furthermore, the behavioral alterations of bacteria affected by l-fucose were directly reveled by measuring the cell distribution of bacteria swimming near surfaces and bacterial surface adhesion, suggesting that l-fucose promotes bacterial surface aggregation and surface adhesion. The effect of l-fucose on bacterial swimming motility characterized in this study are consistent with the key role that l-fucose plays in bacterial colonization.

Solubilization of luteolin in PVP40 solid dispersion improves inflammation-induced insulin resistance in mice.

Our previous studies have confirmed that luteolin (LU) has a good therapeutic effect on obesity and its complications. However, due to its poor water solubility, the bioavailability is low with limited clinical application. Therefore, the water-soluble solid dispersions (SD) of luteolin were prepared with polyvinylpyrrolidone (PVP) (K10, K40 & K90) by solvent evaporation. The polyvinylpyrrolidone K40 (PVP40) was selected as the ideal carrier to formulate polyvinylpyrrolidone K40-luteolin solid dispersion (PVP40-LU SD), thereby the solubility of luteolin increased about 250 times compared to the pure luteolin, without changing its physical stability and activity. The crystallinity of luteolin was reduced after the formation of solid dispersion, and no strong drug-polymer interactions were observed. This prepared water-soluble luteolin inhibits the polarization of inflammatory macrophages by decreasing the expression of pro-inflammatory cytokine genes interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in vitro. Moreover, it can improve glucose tolerance and insulin sensitivity quickly after intraperitoneal injection in mice.

Moderate l-lactate administration suppresses adipose tissue macrophage M1 polarization to alleviate obesity-associated insulin resistance.

As a crucial metabolic intermediate, l-lactate is involved in redox balance, energy balance, and acid-base balance in organisms. Moderate exercise training transiently elevates plasma l-lactate levels and ameliorates obesity-associated type 2 diabetes. However, whether moderate l-lactate administration improves obesity-associated insulin resistance remains unclear. In this study, we defined 800 mg/kg/day as the dose of moderate l-lactate administration. In mice fed with a high-fat diet (HFD), moderate l-lactate administration for 12 weeks was shown to alleviate weight gain, fat accumulation, and insulin resistance. Along with the phenotype alterations, white adipose tissue thermogenesis was also found to be elevated in HFD-fed mice. Meanwhile, moderate l-lactate administration suppressed the infiltration and proinflammatory M1 polarization of adipose tissue macrophages (ATMs) in HFD-fed mice. Furthermore, l-lactate treatment suppressed the lipopolysaccharide-induced M1 polarization of bone marrow-derived macrophages (BMDMs). l-lactate can bind to the surface receptor GPR132, which typically drives the downstream cAMP-PKA signaling. As a nutrient sensor, AMP-activated protein kinase (AMPK) critically controls macrophage inflammatory signaling and phenotype. Thus, utilizing inhibitors of the kinases PKA and AMPK as well as siRNA against GPR132, we demonstrated that GPR132-PKA-AMPKα1 signaling mediated the suppression caused by l-lactate treatment on BMDM M1 polarization. Finally, l-lactate addition remarkably resisted the impairment of lipopolysaccharide-treated BMDM conditional media on adipocyte insulin sensitivity. In summary, moderate l-lactate administration suppresses ATM proinflammatory M1 polarization through activation of the GPR132-PKA-AMPKα1 signaling pathway to improve insulin resistance in HFD-fed mice, suggesting a new therapeutic and interventional approach to obesity-associated type 2 diabetes.

Global RNA editing identification and characterization during human pluripotent-to-cardiomyocyte differentiation.

RNA editing is widely involved in stem cell differentiation and development; however, RNA editing events during human cardiomyocyte differentiation have not yet been characterized and elucidated. Here, we identified genome-wide RNA editing sites and systemically characterized their genomic distribution during four stages of human cardiomyocyte differentiation. It was found that the expression level of ADAR1 affected the global number of adenosine to inosine (A-to-I) editing sites but not the editing degree. Next, we identified 43, 163, 544, and 141 RNA editing sites that contribute to changes in amino acid sequences, variation in alternative splicing, alterations in miRNA-target binding, and changes in gene expression, respectively. Generally, RNA editing showed a stage-specific pattern with 211 stage-shared editing sites. Interestingly, cardiac muscle contraction and heart-disease-related pathways were enriched by cardio-specific editing genes, emphasizing the connection between cardiomyocyte differentiation and heart diseases from the perspective of RNA editing. Finally, it was found that these RNA editing sites are also related to several congenital and noncongenital heart diseases. Together, our study provides a new perspective on cardiomyocyte differentiation and offers more opportunities to understand the mechanisms underlying cell fate determination, which can promote the development of cardiac regenerative medicine and therapies for human heart diseases.

Measurements of Chemical Compositions in Corn Stover and Wheat Straw by Near-Infrared Reflectance Spectroscopy.

Rapid, non-destructive methods for determining the biochemical composition of straw are crucial in ruminant diets. In this work, ground samples of corn stover (n = 156) and wheat straw (n = 135) were scanned using near-infrared spectroscopy (instrument NIRS DS2500). Samples were divided into two sets, with one set used for calibration (corn stover, n = 126; wheat straw, n = 108) and the remaining set used for validation (corn stover, n = 30; wheat straw, n = 27). Calibration models were developed utilizing modified partial least squares (MPLS) regression with internal cross validation. Concentrations of moisture, crude protein (CP), and neutral detergent fiber (NDF) were successfully predicted in corn stover, and CP and moisture were in wheat straw, but other nutritional components were not predicted accurately when using single-crop samples. All samples were then combined to form new calibration (n = 233) and validation (n = 58) sets comprised of both corn stover and wheat straw. For these combined samples, the CP, NDF, and ADF were predicted successfully; the coefficients of determination for calibration (RSQC) were 0.9625, 0.8349, and 0.8745, with ratios of prediction to deviation (RPD) of 6.872, 2.210, and 2.751, respectively. The acid detergent lignin (ADL) and moisture were classified as moderately useful, with RSQC values of 0.7939 (RPD = 2.259) and 0.8342 (RPD = 1.868), respectively. Although the prediction of hemicellulose was only useful for screening purposes (RSQC = 0.4388, RPD = 1.085), it was concluded that NIRS is a suitable technique to rapidly evaluate the nutritional value of forage crops.

Chrysin Stimulates Subcutaneous Fat Thermogenesis in Mice by Regulating PDGFRα and MicroRNA Expressions.

The activation of adipose tissue browning and thermogenesis provides a new strategy to counter obesity and associated metabolic diseases. Here, a natural flavonoid chrysin is used as the supplement of a high-fat diet (HFD). Dietary chrysin alleviates adiposity and insulin resistance in HFD-fed mice. Meanwhile, dietary chrysin elevates systemic energy expenditure and enhances the uncoupling protein-1 (UCP1) level in subcutaneous adipose tissue (SAT), which is accompanied by the increased thermogenic program, beige preadipocyte number, and angiogenesis in SAT. Dietary chrysin also induces the expression of SAT platelet-derived growth factor receptor α (PDGFRα), which commits adipose progenitor cells to differentiate into beige or white adipocytes in response to various environmental signals. Double immunofluorescent staining for UCP1 and PDGFRα reveals that chrysin elevates the number of UCP1+PDGFRα+ beige progenitors in SAT. Further, chrysin treatment reverses the effects of the specific PDGFRα inhibitor imatinib on browning differentiation of stromal vascular fraction cells from SAT. Finally, chrysin-induced adipocyte browning is correlated with the expressions of microRNAs as PDGFRα inhibitors or thermogenesis suppressors. In conclusion, dietary chrysin promotes subcutaneous adipocyte browning and systematic energy expenditure by regulating PDGFRα and microRNA expressions in HFD-fed mice.

In situ growth of ZIF-8 on gold nanoparticles/magnetic carbon nanotubes for the electrochemical detection of bisphenol A.

We herein report a facile and scalable strategy for the fabrication of a metal-organic framework (MOF) based composite by in situ growing ZIF-8 on gold nanoparticle (AuNP) loaded magnetic carbon nanotubes (mCNTs). AuNPs were firstly loaded on PEI (polyethylenimine) modified mCNTs by electrostatic forces, and then AuNPs/mCNTs were encapsulated into the ZIF-8 frame through in situ self-assembling of zinc ions and 2-methylimidazole. The morphology, spectroscopy and structural properties of the AuNP/mCNT@ZIF-8 nanocomposites were systematically characterized. The conductivity-strain tests revealed that the in situ insertion of AuNPs/mCNTs in ZIF-8 could not only shorten the electron transfer distance between active sites and mCNTs, but also increase the dispersion of mCNTs, which would benefit the electron and mass transfer. Besides, by adopting the AuNP/mCNT@ZIF-8 nanocomposite-modified glassy carbon electrode (GCE) as the working electrode, a novel electrochemical sensor was successfully developed for the detection of bisphenol A (BPA). A linear range of BPA detection from 1 µM to 100 µM with a limit of detection of 690 nM was favorably obtained. Moreover, the developed sensor exhibited satisfactory reproducibility and superior stability with excellent anti-interference ability, and was successfully applied in the detection of BPA in real samples.

AIEgen modulated per-functionalized flower-like IRMOF-3 frameworks with tunable light emission and excellent sensing properties.

A series of functional IRMOF-3 frameworks with solid-state luminescence and tuneable light emission (from 490 to 608 nm) have been synthesized by per-functionalizing AIE-active Schiff-bases with zinc. These precursor AIE-active ligands endowed the functional frameworks with boosted fluorescence emission efficiencies (from 0.16% to 1.03%). IRMOF-3-h revealed a flower-like morphology attributed to the formation of J-aggregates, and could be used as a fluorescent probe for sensitive detection of copper(ii) (135 pM) and thiols (subnanomole).

Metformin Alters the Chemotaxis and Flagellar Motility of Escherichia coli.

Metformin is a biguanide molecule that is widely prescribed to treat type 2 diabetes and metabolic syndrome. Although it is known that metformin promotes the lifespan by altering intestinal microorganism metabolism, how metformin influences and alters the physiological behavior of microorganisms remains unclear. Here we studied the effect of metformin on the behavior alterations of the model organism Escherichia coli (E. coli), including changes in chemotaxis and flagellar motility that plays an important role in bacterial life. It was found that metformin was sensed as a repellent to E. coli by tsr chemoreceptors. Moreover, we investigated the chemotactic response of E. coli cultured with metformin to two typical attractants, glucose and α-methyl-DL-aspartate (MeAsp), finding that metformin prolonged the chemotactic recovery time to the attractants, followed by the recovery time increasing with the concentration of stimulus. Metformin also inhibited the flagellar motility of E. coli including the flagellar motor rotation and cell swimming. The inhibition was due to the reduction of torque generated by the flagellar motor. Our discovery that metformin alters the behavior of chemotaxis and flagellar motility of E. coli could provide potential implications for the effect of metformin on other microorganisms.

Characterization of chemotaxis and motility response towards fructose in Escherichia coli.

Peritrichously flagellated bacteria such as Escherichia coli (E. coli) perform chemotaxis by a biased random walk toward various chemicals, which was driven by the bacterial flagellar motor. Fructose, a typical monosaccharide that can attract E. coli. However, little is known about the chemotaxis and motility response of E. coli towards fructose. Here, we characterized the chemotaxis behavior of E. coli to different concentrations of fructose from 0 mM to 50 mM by using microfluidics and bead assay. We observed the wild-type cells responded to the stimulus of fructose, which suggested fructose is an attractant to E. coli, while the cells defective in chemotaxis could not sense the stimulus of fructose. The motility of wild-type cells was reduced in various concentrations of fructose, which helped the aggregation of cells near surfaces, in contrast with the result that the fructose showed no effect on the motility of the cells defective in chemotaxis. Similar phenomena are expected to be found in the effect of other monosaccharides to E. coli.

Genomic Identification of RNA Editing Through Integrating Omics Datasets and the Clinical Relevance in Hepatocellular Carcinoma.

RNA editing is a widespread post-transcriptional mechanism to introduce single nucleotide changes to RNA in human cancers. Here, we characterized the global RNA editing profiles of 373 hepatocellular carcinoma (HCC) and 50 adjacent normal liver samples from The Cancer Genome Atlas (TCGA) and revealed that most editing events tend to occur in minor percentage of samples with moderate editing degrees (20-30%). Moreover, these RNA editing prefer to be A-to-I RNA editing in protein coding genes, especially in 3'UTR regions. Considering the association between DNA mutation and RNA editing, our analysis found that RNA editing maybe a complementary event for DNA mutation of HCC risk genes in HCC patients. We next identified 454 HCC-related editing sites, and many locate on the same genes with the same editing patterns. The functional consequences of editing revealed 2,086 functional editing sites and demonstrated that most editing in coding regions are non-synonymous variations. Furthermore, our results showed that editing in the 3'UTR regions tend to influence miRNA-target binding, and the editing degree seems to be negatively correlated with gene expression. Finally, we found that 46 HCC-related editing sites with consequence are able to distinguish the prognosis differences of HCC patients, suggesting their clinical relevance. Together, our results highlight RNA editing as a valuable molecular resource for investigating HCC mechanisms and clinical treatments.

Cumulative risk assessment of phthalates in edible vegetable oil consumed by Chinese residents.

BACKGROUND: Phthalates have been widely used as plasticizers in various industries and are widely focused on in the international community as a result of their reproductive toxicity. Exposure of Chinese residents to phthalates via edible vegetable oil occurs often. In the present study, gas chromatography-mass spectrometry was used to detect the two main phthalates bis(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) in four major edible vegetable oil sources: an edible oil blend, soybean oil, peanut oil and rapeseed oil (a total of 1016 samples), as collected throughout China. Furthermore, cumulative risk assessment was used to estimate the reproductive health risk to Chinese residents caused by the phthalates that come from edible vegetable oils. RESULTS: Both phthalates were detected in four major edible vegetable oil sources. The phthalate with the highest detection rate was DBP (13.48%), followed by DEHP (7.78%). The results of the cumulative risk assessment showed that the hazard indices of these two phthalates in edible vegetable oils were less than 1, except in soybean oil. Nevertheless, the two phthalates had the lowest detection rates in soybean oil, which were 1.94% (DEHP) and 5.16% (DBP). In China, contamination levels of phthalates in the soils where oil crops are cultivated have a great influence on the phthalate concentrations in edible vegetable oils. CONCLUSION: It is recommended that Chinese residents who are consuming soybean oil choose well-known brands and regularly change their brand of consumption. The phthalates in edible vegetable oils pose a relatively small reproductive health risk to Chinese residents. © 2019 Society of Chemical Industry.

Public risk perception and willingness to mitigate climate change: city smog as an example.

Climate change harms people's health and ecosystems. Encouraging the public to adopt behaviors that help to combat climate change can, at the same time, contribute to efforts to control and solve related serious environmental problems. This study aims to ascertain the way in which the public perceives risks related to climate change and adopts behaviors to respond to the issue. Using city smog as an example, this study proposes a conceptual model that integrates the theory of planned behavior (TPB), smog knowledge and risk perception. It aims to elucidate determinants of smog-reduction behavior. Data were obtained through questionnaire surveys. The results confirm the relationships among the core variables of the TPB and risk perception. Firstly, they confirm that TPB is an effective model for predicting responses to city smog, and secondly, they show that risk perception is significantly positive in predicting attitude and behavioral intention. In addition, our analysis confirms that knowledge about smog is a positive antecedent variable in risk perception, attitude, and perceived behavioral control. The paper contributes to the extension of the TPB model and to the enrichment of its application in the context of city smog. It also has practical implications both for people experiencing city smog, and for authorities such as local governments and environmental organizations. Governments and organizations need to make efforts to spread information concerning the harmful effects of city smog, because in doing so they can strengthen people's intention to participate in smog-reduction behavior.

The Second Messenger c-di-GMP Adjusts Motility and Promotes Surface Aggregation of Bacteria.

Bacteria can use the second messenger c-di-GMP to adjust their motility in response to environmental cues. The protein YcgR, upon binding of c-di-GMP, interacts with the flagellar motor to affect the motor behavior. However, the full feature of the effects of c-di-GMP::YcgR on the flagellar motor remains unclear, and its interacting partners on the motor is still controversial. Here, we characterized the effects of c-di-GMP::YcgR on the torque-speed curve of the flagellar motor, one of the most important properties of the motor, finding that it affects the motor behavior throughout the full range of load conditions from zero to high loads by shifting the motor torque-speed curve downward. We also investigated the interacting partner on the motor through dynamical fluorescent studies, finding that c-di-GMP::YcgR mainly interacts with the motor-switch complex instead of the torque-generating units (stators). To directly test the behavioral consequence of elevated c-di-GMP levels, we measured the distribution of bacteria swimming near a surface, finding that elevated c-di-GMP levels promote bacterial aggregation on surfaces. The effects of c-di-GMP on bacterial motile behavior that we characterized here are consistent with the key role that c-di-GMP plays in the transition between motile and sedentary forms of bacterial life.

Effects of early feeding on the host rumen transcriptome and bacterial diversity in lambs.

Early consumption of starter feed promotes rumen development in lambs. We examined rumen development in lambs fed starter feed for 5 weeks using histological and biochemical analyses and by performing high-throughput sequencing in rumen tissues. Additionally, rumen contents of starter feed-fed lambs were compared to those of breast milk-fed controls. Our physiological and biochemical findings revealed that early starter consumption facilitated rumen development, changed the pattern of ruminal fermentation, and increased the amylase and carboxymethylcellulase activities of rumen micro-organisms. RNA-seq analysis revealed 225 differentially expressed genes between the rumens of breast milk- and starter feed-fed lambs. These DEGs were involved in many metabolic pathways, particularly lipid and carbohydrate metabolism, and included HMGCL and HMGCS2. Sequencing analysis of 16S rRNA genes revealed that ruminal bacterial communities were more diverse in breast milk-than in starter feed-fed lambs, and each group had a distinct microbiota. We conclude that early starter feeding is beneficial to rumen development and physiological function in lambs. The underlying mechanism may involve the stimulation of ruminal ketogenesis and butanoate metabolism via HMGCL and HMGCS2 combined with changes in the fermentation type induced by ruminal microbiota. Overall, this study provides insights into the molecular mechanisms of rumen development in sheep.