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Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells (BMEC). The objectives of this study were to determine how glucose affects hexokinase (HK) activity in BMEC and investigate the regulatory effect of HK in kappa casein (CSN3) synthesis via the mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway in BMEC. For this, HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 system. The gene and protein expression, glucose uptake, and cell proliferation were measured. We found that glucose uptake, cell proliferation, CSN3 gene expression levels, and expression of HK1 and HK2 increased with increasing glucose concentrations. Notably, glucose uptake was significantly reduced in HK2 knockout (HK2KO) BMEC treated with 17.5 mM glucose. Moreover, under the same glucose treatment conditions, the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout (HK1KO) and HK2KO BMEC compared with that in wild-type BEMC. We further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1 (S6K1) were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM glucose. As expected, the levels of glucose-6-phosphate and the mRNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose treatment. These results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment, which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.

In this study, the deformation and damage mechanisms of a high-concentration Re/Ru single-crystal nickel-based alloy at ultrahigh temperatures are studied via creep performance tests, microstructural observations and dislocation configuration diffraction contrast analyses at 1170 °C. The results show that the alloy has good mechanical properties. In addition, the creep life is 155 h at 1170 °C and 120 MPa. During creep, dislocations moving in the γ matrix can react with each other to form a dense and complete dislocation network. This dense and complete dislocation network can effectively prevent dislocations from shearing into the γ′ phase and reduce the creep rate during the steady-state creep period. In the late stage of ultrahigh-temperature creep, stacking faults are found in the γ′ phase, indicating that the alloy has a low stacking fault energy and that the stacking faults that form in this alloy can serve as obstacles for dislocation movement. In addition, the dislocations that cut into the γ′ phase in the late stage of ultrahigh temperature creep can cross-slip from the {111} plane to the {100} plane, forming a KW lock, which can inhibit the slip and cross-slip dislocation movement modes. Therefore, the presence of dislocation networks, stacking faults and KW locks in alloys is the main reason for the relatively low strain rate and good creep resistance of these alloys under ultrahigh-temperature creep conditions. In the late stage of creep, the initiated dislocations slip in two orientations, resulting in kinking of the rafted γ′ phase and crack initiation at the top interface of the twisted rafted γ/γ′ two phases. Under the action of shear stress, the crack generates tensile stress perpendicular to the stress axis, and it expands in this direction. This crack is the damage mechanism of the alloy during ultrahigh-temperature creep.

Early weaning leads to weaning stress in calves, which hinders healthy growth and development. As an excellent sweetener applied in food, steviol glycosides (STE) has also been shown to exhibit positive biological activity in monogastric animals. Therefore, this study aimed to evaluate the impact of incorporating STE as a dietary supplement on rumen development, fermentation, and microbiota of rumen in weaned calves. This study selected 24 healthy Holstein bull calves and randomly allocated them into two groups (CON and STE). The results indicated that supplementation STE group improved rumen development in weaned calves, as demonstrated by a marked increase in the weight of the rumen, as well as the length and surface area of the rumen papilla. Compared with the CON group, the concentrations of total volatile fatty acids (TVFA), propionate, butyrate, and valerate were higher in the STE group. Moreover, STE treatment increased the relative abundance of Firmicutes and Actinobacteria at the phylum level. At the genus level, the STE group showed a significantly increased relative abundance of Succiniclasticum, Lachnospiraceae_NK3A20_group, and Olsenella, and a decreased relative abundance of Acinetobacter compared to the CON group. Pusillimonas, Lachnospiraceae_NK3A20_group, Olsenella, and Succiniclasticum were significantly enriched in rumen chyme after supplementation with STE, as demonstrated by LEfSe analysis. Overall, our findings revealed that rumen bacterial communities altered in response to the dietary supplementation with STE, and some bacterial taxa in these communities may have positive effects on rumen development during this period.

In recent years, negative Poisson's ratio structures have attracted many scholars' attention as a result of their distinctive mechanical properties and exceptional energy absorption ability. However, the geometry of structures with negative Poisson's ratio is generally complex, and the preparation of the model has become problematic. Therefore, the increasingly mature additive manufacturing industry can be used to prepare three-dimensional models. In this paper, five negative Poisson's ratio structures were fabricated by 3D printing, and cementitious materials were poured onto them. The purpose of this study is to investigate the mechanical properties of different structural reinforced concrete specimens under in-plane quasi-static uniaxial compression and obtain the deformation and failure modes of different structural reinforced concrete specimens. In the experiment, two-dimensional digital image correlation (2D-DIC) and acoustic emission (AE) techniques were used to obtain plane displacement and internal damage evolution, and the plane strain field was calculated using Vic-2D software. The experimental results show that the ductility and energy absorption capacity of structural reinforced concrete specimens have been significantly improved, and there are significant differences in the performance of different structural reinforced concrete due to different deformation modes.

The objectives of the current study were to evaluate the fluctuations in production performance, rumen fermentation, and microbial community in lactating dairy cows fed a high-grain diet (HG). In this study, 16 healthy Holstein lactating dairy cattle with similar milk yields of 16.80 ± 4.30 kg/d, days in milk 171.44 ± 23.25 days, and parity 2.2 ± 1.5 times were selected and randomly allocated into two groups. One group was fed a low-grain diet (LG; 40% concentrate, DM basis; n = 8), and the other group was fed a high-grain diet (HG; 60% concentrate, DM basis; n = 8). The experiment lasted 6 weeks, including 1 week for adaptation. The experimental results showed that the milk fat content in the milk of lactating cows in the HG group was significantly reduced (p < 0.05), and the milk urea nitrogen (MUN) content showed an increasing trend (0.05 < p < 0.10) compared with the LG group. Compared with the LG group, rumen fluid pH was significantly decreased after feeding a high-grain diet, and contents of total volatile fatty acids (TVFA), acetate, propionate, and butyrate were significantly increased (p < 0.05). The acetate/propionate significantly decreased (p < 0.05). HG group significantly increased the abundance of Prevotella and Bacteroides in rumen fluid while significantly reducing the abundance of Methanobrevibacter and Lachnospiraceae ND3007_group (p < 0.05). Microorganisms with LDA scores > 2 were defined as unique, with the bacterial genus Anaerorhabdus_furcosa_group identified as a biomarker for the LG group, and the unique bacterial genus in the HG group were Prevotella, Stenotrophomonas, and Xanthomonadaceae. The prediction results of microbial function showed that a total of 18 KEGG differential pathways were generated between the two treatment groups, mainly manifested in metabolic pathways, signal transduction, and the immune system. In conclusion, the HG group promoted rumen fermentation by altering the microbial composition of lactating cows. Our findings provide a theoretical basis for the rational use of high-grain diets to achieve high yields in intensive dairy farming.

Acute diarrhoea and intestinal inflammation represent one of the most prevalent clinical disorders of milk production, resulting in enormous annual financial damage for the dairy sector. In the context of an unsatisfactory therapeutic effect of antibiotics, the natural products of plants have been the focus of research. Quercetin is an important flavonoid found in a variety of plants, including fruits and vegetables, and has strong anti-inflammatory effects, so it has received extensive attention as a potential anti-inflammatory antioxidant. However, the underlying basis of quercetin on inflammatory reactions and oxidative tension generated by lipopolysaccharide (LPS) in bovine intestinal epithelial cells (BIECs) is currently unexplained. This research aimed to determine the influence of quercetin on LPS-induced inflammatory reactions, oxidative tension, and the barrier role of BIECs. Our findings demonstrated that BIEC viability was significantly improved in LPS-treated BIEC with 80 μg/mL quercetin compared with the control group. Indicators of oxidative overload and genes involved in barrier role revealed that 80 μg/mL quercetin efficiently rescued BIECs from oxidative and barrier impairment triggered by 5 μg/mL LPS. In addition, the mRNA expression of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6, as well as chemokines CXCL2, CXCL5, CCL5, and CXCL8, was diminished in LPS-treated BIECs with 80 μg/mL quercetin compared with LPS alone. Furthermore, the mRNA expression of toll-like receptor 4 (TLR4), CD14, myeloid differential protein-2 (MD2), and myeloid differentiation primary response protein (MyD88) genes associated with the TLR4 signal mechanism was markedly reduced by the addition of quercetin to LPS-modulated BIECs, indicating that quercetin can suppress the TLR4 signal mechanism. We performed Western blotting on the NF-κB signalling mechanism and compared it with immunofluorescence to further corroborate this conclusion. The LPS treatment enhanced the proportions of p-IκBα/GAPDH and p-p65/GAPDH. Compared with the LPS-treated group, quercetin administration decreased the proportions of p-IκBα/GAPDH and p-p65/GAPDH. In addition, immunofluorescence demonstrated that quercetin greatly reduced the LPS-induced nuclear translocation of NF-κB p65 in BIECs. The benefits of quercetin on inflammatory reactions in LPS-induced BIECs may be a result of its capacity to inhibit the TLR4-mediated NF-κB signalling mechanism. These findings suggest that quercetin can be used as an anti-inflammatory reagent to treat intestinal inflammation induced by LPS release.

This experiment investigated the effects of different levels of bile acid (BA) additives in diets on the lactation performance, serum antioxidant metabolites, and serum biochemical indices of 60 multiparous mid-lactation dairy cows. The cows were randomized to receive one of the four homogeneous treatments, with the BA preparation supplemented at 0, 6, 12, and 18 g/head/d. The experiment lasted for 14 weeks. The first 2 weeks were the pre-feeding period. The milk yield and composition data were recorded weekly, and the dry matter intake and antioxidative blood index were analyzed on the 6th, 10th, and 14th weeks of the study. On the 84th day of the experiment, the experimental group exhibited significantly higher levels of total protein and albumin, by 57.5% and 55.6%, respectively, compared to the control group (p < 0.05). On both the 28th and 84th days of the trial, the experimental group showed a markedly higher lipase content compared to the control group, by 26.5% and 25.2%, respectively (p < 0.05). Furthermore, the experimental group displayed notably elevated levels of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity, surpassing the control group by 17.4%, 21.6%, and 8.7%, respectively. In conclusion, BA additives improve the serum antioxidant indices of dairy cows, thereby enhancing the performance of these cows.

Abstract Background To validate tumor volume-based imaging markers for predicting local recurrence-free survival (LRFS) in locoregionally advanced nasopharyngeal carcinoma patients, who underwent induction chemotherapy followed by definitive intensity-modulated radiotherapy. Methods We enrolled 145 patients with stage III–IVA nasopharyngeal carcinoma in this retrospective study. Pre-treatment tumor volume (Vpre) and late-course volume (LCV) were measured based on the MRIs scanned before treatment and during the first 3 days in the sixth week of radiotherapy, respectively. The volume regression rate (VRR) was calculated according to Vpre and LCV. Receiver operating characteristic (ROC) curves were used to identify the cut-off best separating patient subgroups in assessing the prognostic value of Vpre, LCV and VRR. The Kaplan–Meier method was used for survival analysis. Prognostic analyses were performed using univariate and multivariate COX proportional hazard models. Results The LCV was 5.3 ± 0.5 (range 0–42.1) cm3; The VRR was 60.4 ± 2.2% (range 2.9–100.0). The median follow-up period was 36 months (range 6–98 months). The cut-off value of LCV determined by the ROC was 6.8 cm3 for LRFS prediction (sensitivity 68.8%; specificity 79.8%). The combination of LCV and VRR for LRFS prediction (AUC = 0.79, P 6.8 cm3 . Multivariate Cox regression showed LCV was the only independent prognostic factor for local control (HR = 7.80, 95% CI 2.69–22.6, P

Glucose and amino acids are important sources of nutrients in the synthetic milk of dairy cows, and understanding the fate of amino acids is essential to optimize the utilization of amino acids in milk protein synthesis, thereby reducing nutrient inefficiencies during lactation. The purpose of this study was to investigate the effects of LPS and different concentrations of glucose on (1) the expression of inflammatory factors and genes, (2) the glucose metabolism, and (3) amino acid utilization in BMECs. The results showed that there was an interaction (LPS × glucose, p < 0.05) between LPS and glucose content in the inflammatory cytokine genes (IL-6 and TNF-α) and the inflammatory regulatory genes (CXCL2, CXCL8, and CCL5). With the addition of LPS, the HG + LPS group caused downregulated (p < 0.05) expression of IL-6 and TNF-α, compared with the LG + LPS group. Interestingly, compared with the LG + LPS group, the HG + LPS group upregulated (p < 0.05) the expression of CXCL2, CXCL8, and CCL5. LPS supplementation increased (p = 0.056) the consumption of glucose and GLUT1 gene expression (p < 0.05) and tended to increase (p = 0.084) the LDHA gene expression of BMECs under conditions of different concentrations of glucose culture. High glucose content increased (p < 0.001) the consumption of glucose and enhanced (p < 0.05) the GLUT1, HK1, HK2, and LDHA gene expression of BMECs with or without LPS incubation, and there was an interaction (LPS × glucose, p < 0.05) between LPS and glucose concentrations in GLUT1 gene expression. In this study, LPS enhanced (p < 0.05) the consumption of amino acids such as tryptophan, leucine, isoleucine, methionine, valine, histidine, and glutamate, while high levels of glucose decreased (p < 0.01) consumption, except in the case of tyrosine. For histidine, leucine, isoleucine, and valine consumption, there was an interaction (LPS × glucose, p < 0.05) between LPS and glucose levels. Overall, these findings suggest that relatively high glucose concentrations may lessen the LPS-induced BMEC inflammatory response and reduce amino acid consumption, while low glucose concentrations may increase the demand for most amino acids through proinflammatory responses.

The fatty acid profiles of ruminant-derived products are closely associated with human health. Ruminal microbiota play a vital role in modulating rumen biohydrogenation (BH). The aim of this study was to assess the influence of dietary supplementation with phlorotannins (PTs) extracted from Sargassum on rumen fermentation, fatty acid composition and bacterial communities by an in vitro culture study. The inclusion of PTs in the diet increased dry matter digestibility and gas production, and reduced ammonia-N concentration and pH. PT extract inhibited rumen BH, increasing the content of trans-9 C18:1, cis-9 C18:1, trans-9 and trans-12 C18:2 and reducing C18:0 concentration. 16S rRNA sequencing revealed that PTs caused an obvious change in rumen bacterial communities. The presence of Prevotella decreased while carbohydrate-utilizing bacteria such as Prevotellaceae_UCG-001, Ruminococcus, Selenomonas, Ruminobacter and Fibrobacter increased. Correlation analysis between rumen FA composition and the bacterial microbiome revealed that Prevotellaceae_UCG-001, Anaerovorax, Ruminococcus, Ruminobacter, Fibrobacter, Lachnospiraceae_AC2044_group and Clostridia_UCG-014 might have been involved in the BH process. In conclusion, the results suggest that the inclusion of PTs in the diet improved rumen fermentation and FA composition through modulating the rumen bacterial community.

This study investigated the impact of dietary neutral detergent fiber (NDF) levels (25.49%, 28.65%, 31.66%, and 34.65%, respectively) on the feeding behavior, rumen fermentation, cellulolytic bacteria, and production performance of dairy cows during peak lactation. A feeding experiment was conducted using four fistulated Holstein dairy cows (600 ± 25 kg) with days in milk (50 ± 15 days), employing a 4 × 4 Latin square design to assign the cows to four groups. The results demonstrated that increasing NDF levels in the diet had the following effects: (1) A linear decrease in dry matter intake (DMI), NDF intake, and physically effective NDF8.0 (peNDF8.0) intake; a linear increase in the average time spent eating and ruminating, as well as the time spent eating and ruminating per kilogram of dry matter (DM); a quadratic response in the time spent ruminating per kilogram of NDF and peNDF8.0. (2) A linear increase in average pH value, acetate concentration, and the proportions of Fibrobacter succinogenes and Ruminococcus flavefaciens among total bacteria; a linear decrease in ammonia nitrogen (NH3-N) concentration, microbial crude protein (MCP), total volatile fatty acid (TVFA), propionate, butyrate, and lactate. (3) A linear decrease in milk yield, milk protein percentage, and nitrogen efficiency of dairy cows; a linear increase in milk fat percentage and milk urea nitrogen (MUN) concentration. Based on the combined results, it was found that diets with 25% and 34% NDF had detrimental effects on the feeding behavior, rumen fermentation, and production performance of dairy cows. However, the diet with 28% NDF showed superior outcomes in production performance compared to the one with 31% NDF. Therefore, it is strongly recommended to include a diet containing 28% NDF during the critical peak lactation period for dairy cows.

Through creep measurement and microstructure observation, the microstructure evolution and deformation mechanism of a [111]-oriented nickel-based single-crystal superalloy at 980 °C/186 MPa were studied. The results show that the [111]-oriented single-crystal superalloy after full heat treatment featured cubic γ' phases coherently embedded in the γ phases and regularly arranged along the direction. During the creep, the cubic γ' phases were interconnected, phase thickening and coarsening occurred, and the phase transformed into a lamellar rafted structure that grew staggered in three-dimensional space. The deformation characteristics of the [111]-oriented nickel-based single-crystal superalloy during creep under experimental conditions were that the dislocations slipped, propagated, and climbed over the γ' phases in the matrix, and then cut into the γ' phase and cross-slipped from the {111} plane to the {100} plane to form Kear–Wilsdorf (K–W) locks. Finally, the dislocations cut the γ' phase in the form of dislocation pairs, causing the γ' phase to lose its creep resistance, resulting in creep fracture.

The primary product of rumen fermentation is acetic acid, and its sodium salt is an excellent energy source for post-partum cows to manage negative energy balance (NEB). However, it is unknown how adding sodium acetate (NAc) may affect the rumen bacterial population of post-partum cows. Using the identical nutritional total mixed ration (TMR), this research sought to characterize the impact of NAc supplementation on rumen fermentation and the composition of bacterial communities in post-partum cows. After calving, 24 cows were randomly assigned to two groups of 12 cows each: a control group (CON) and a NAc group (ACE). All cows were fed the same basal TMR with 468 g/d NaCl added to the TMR for the CON group and 656 g/d NAc added to the TMR for the ACE group for 21 days after calving. Ruminal fluid was collected before morning feeding on the last day of the feeding period and analyzed for rumen bacterial community composition by 16S rRNA gene sequencing. Under the identical TMR diet conditions, NAc supplementation did not change rumen pH but increased ammonia nitrogen (NH3-N) levels and microbial crude protein (MCP) concentrations. The administration of NAc to the feed upregulated rumen concentrations of total volatile fatty acids (TVFA), acetic, propionic, isovaleric and isobutyric acids without affecting the molar ratio of VFAs. In the two experimental groups, the Bacteroidota, Firmicutes, Patescibacteria and Proteobacteria were the dominant rumen phylum, and Prevotella was the dominant rumen genus. The administration of NAc had no significant influence on the α-diversity of the rumen bacterial community but upregulated the relative abundance of Prevotella and downregulated the relative abundance of RF39 and Clostridia_UCG_014. In conclusion, the NAc supplementation in the post-peripartum period altered rumen flora structure and thus improved rumen fermentation in dairy cows. Our findings provide a reference for the addition of sodium acetate to alleviate NEB in cows during the late perinatal period.

This paper presents a machine learning approach to localizing a five-peak narrow-band modulated sinusoidal signal excitation source within a composite panel. In particular, the Back Propagation (BP) neural network is used. The idea is to use the arrival time of the first wave packet in a five-peak wave signal to locate their source. Specifically, this paper divides the composite material board into multiple regions, designs 8 receiving points to receive the signal from the excitation source, and finds the region where each source is located. The COMSOL numerical simulation platform is used to build a composite plate model and simulate the propagation of five-peak waves to train and test the machine learning network. Correspondingly, carry out experimental verification and use a scanning laser Doppler vibrometer (SLDV) to build a non-contact experimental platform to obtain the wave field information in the composite material plate. The results show that BP neural networks can learn to map signal features to their sources in both contexts.

Bovine liver mainly utilizes the propionate as a gluconeogenic substrate to synthesize the glucose. However, the mechanism underlying the regulatory effects of propionate on the glucose production in bovine hepatocytes remains less known. Previous studies have demonstrated G protein-coupled receptor 41 (GPR41) as receptors for propionate. We hypothesized that propionate may regulate the glucose production by GPR41 in bovine hepatocytes. Therefore, the aim of the study was to investigate the regulatory effects of propionate and GPR41 on glucose production in bovine hepatocytes. Hepatocytes with GPR41 overexpression were incubated in the presence of either 0 or 3 mM propionate for 24 h. These results showed that the expression of phosphoenolpyruvate carboxykinase 2 (PCK2) and pyruvate carboxylase (PC) genes involved in gluconeogenesis was enhanced (P < 0.01) with propionate treatment. Remarkably, the addition of propionate promotes the glucose production in bovine hepatocytes. Expression of GPR41 was increased by the addition of propionate in bovine hepatocytes overexpressed GPR41 by overexpression plasmid AAV1 compared with the absence of propionate. Interestingly, expression of PCK2 was markedly attenuated in GPR41 overexpressed-hepatocytes with propionate. Importantly, overexpression of GPR41 attenuated glucose output in propionate-induced bovine hepatocytes. These findings revealed that GPR41 negatively regulates glucose production by downregulating the expression of PCK2 in propionate-induced bovine hepatocytes.

Excessive lipid mobilization will snatch cell membrane lipids in postpartum dairy cows, which may impair the function of immune cells, including peripheral mononuclear cells (PBMCs) and polymorphonuclear granulocytes (PMNs). Acetate, as a precursor and the energy source of milk fat synthesis, plays a key role in lipid synthesis and the energy supply of dairy cows. However, there is little information about the effect of sodium acetate (NaAc) on the immune function of PBMC and PMN in postpartum dairy cows. Therefore, this study aimed to evaluate the effects of NaAc on the immune functions of PBMCs and PMNs in postpartum dairy cows. In this experiment, twenty-four postpartum multiparous Holstein cows were randomly selected and divided into a NaAc treatment group and a control group. Our results demonstrated that the dietary addition of NaAc increased (p < 0.05) the number of monocytes and the monocyte ratio, suggesting that these postpartum cows fed with NaAc may have better immunity. These expressions of genes (LAP, XBP1, and TAP) involved in the antimicrobial activity in PBMCs were elevated (p < 0.05), suggesting that postpartum dairy cows supplemented with NaAc had the ability of antimicrobial activity. In addition, the mRNA expression of the monocarboxylate transporters MCT1 and MCT4 in PBMCs was increased (p < 0.05) in diets supplemented with NaAc in comparison to the control. Notably, the expression of the XBP1 gene related to antimicrobial activity in PMN was upregulated with the addition of NaAc. The mRNA expression of genes (TLN1, ITGB2, and SELL) involved in adhesion was profoundly increased (p < 0.05) in the NaAc groups. In conclusion, our study provided a novel resolution strategy in which the use of NaAc can contribute to immunity in postpartum dairy cows by enhancing the ability of antimicrobial and adhesion in PBMCs and PMNs.

This study aimed to assess the effects of partially substituting soybean meal in the diet with slow-release urea (SRU) on the lactation performance, heat shock signal molecules, and environmental sustainability of heat-stressed lactating cows in the middle stage of lactation. In this study, 30 healthy Holstein lactating dairy cattle with a similar milk yield of 22.8 ± 3.3 kg, days in milk of 191.14 ± 27.24 days, and 2.2 ± 1.5 parity were selected and randomly allocated into two groups. The constituents of the two treatments were (1) basic diet plus 500 g soybean meal (SM) for the SM group and (2) basic diet plus 100 g slow-release urea and 400 g corn silage for the SRU group. The average temperature humidity index (THI) during the experiment was 84.47, with an average THI of >78 from day 1 to day 28, indicating the cow experienced moderate heat stress conditions. Compared with the SM group, the SRU group showed decreasing body temperature and respiratory rate trends at 20:00 (p < 0.1). The substitution of SM with SRU resulted in an increasing trend in milk yield, with a significant increase of 7.36% compared to the SM group (p < 0.1). Compared to the SM group, AST, ALT, and γ-GT content levels were significantly increased (p < 0.05). Notably, the levels of HSP-70 and HSP-90α were significantly reduced (p < 0.05). The SRU group showed significantly increased acetate and isovalerate concentrations compared with the SM group (p < 0.05). The prediction results indicate that the SRU group exhibits a significant decrease in methane (CH4) emissions when producing 1 L of milk compared to the SM group (p < 0.05). In summary, dietary supplementation with SRU tended to increase the milk yield and rumen fermentation and reduce plasma heat shock molecules in mid-lactation, heat-stressed dairy cows. In the hot summer, using SRU instead of some soybean meal in the diet alleviates the heat stress of dairy cows and reduces the production of CH4.

Subacute rumen acidosis (SARA) will cause an increase in endotoxin, which will have a negative effect on the bovine rumen epithelial cells (BREC). Flavonoids are effective in treating inflammation caused by endotoxin. Quercetin is a vital flavonoid widely occurring in fruits and vegetables and has received significant interest as a prospective anti-inflammatory antioxidant. Nonetheless, quercetin’s protective machinery against such damage to BREC induced by lipopolysaccharide (LPS) remains unclear. A combined quercetin and LPS-induced BREC inflammation model was utilized to elucidate the effect of quercetin protecting BREC from LPS-induced injury. After treating BREC with different doses of LPS (1, 5, and 10 μg/mL) for 6 h or 24 h, the mRNA expression of inflammatory factors was detected. Our experimental results show the establishment of the BREC inflammation model via mRNA high expression of pro-inflammatory cytokines in BREC following 6 h treatment with 1 µg/mL LPS. The promotive effect of 80 μg/mL quercetin on BREC growth via the cell counting kit-8 (CCK8) assay was observed. The expression of pro-inflammatory cytokines and chemokines, notably tumor necrosis factor α (TNF-α), Interleukin 1β (IL-1β), IL-6, CC-motif chemokine ligand 2 (CCL2), CCL20, CCL28, and CXC motif chemokine 9 (CXCL9), etc., was significantly reduced by quercetin supplementation. We also analyzed the mRNA detection of related pathways by qRT-PCR. Our validation studies demonstrated that quercetin markedly curbed the mRNA expression of the toll-like receptor 4 (TLR4) and myeloid differentiation primary response protein (MyD88) and the nuclear factor-κB (NF-κB) in LPS-treated BREC. In addition, western blot result outcomes confirmed, as expected, that LPS significantly activated phosphorylation of p44/42 extracellular regulated protein kinases (ERK1/2) and NF-κB. Unexpectedly, this effect was reversed by adding quercetin. To complement western blot results, we assessed p-ERK1/2 and p-p65 protein expression using immunofluorescence, which gave consistent results. Therefore, quercetin’s capacity to bar the TLR4-mediated NF-κB and MAPK signaling pathways may be the cause of its anti-inflammatory effects on LPS-induced inflammatory reactions in BREC. According to these results, quercetin may be utilized as an anti-inflammatory medication to alleviate inflammation brought on by high-grain feed, and it also lays out a conceptual foundation regarding the development and utilization of quercetin in the later stage.

Bovine mastitis is one of the most common clinical diseases in dairy cows, causing huge economic losses to the dairy industry. Quercetin is an important flavonoid existing in many food resources, which has attracted widespread attention as a potential anti-inflammatory and antioxidant. However, the molecular mechanism of quercetin on inflammatory responses and oxidative stress in bovine mammary epithelial cells (BMECs) induced by lipopolysaccharide (LPS) remains unknown. The objective of this study was to investigate the effects of quercetin on inflammation responses, oxidative stress, and barrier function of BMEC induced by LPS. Our results showed that BMEC viability was not affected by treatment with 50 and 100 μg/ml of quercetin and 1 μg/ml of LPS compared with control group. The results of oxidative stress indicators and related genes of barrier function indicated that 100 μg/ml of quercetin effectively protected the BMECs from damage of oxidative and barrier induced by 1 μg/ml of LPS. Moreover, the messenger RNA (mRNA) expressions of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and chemokines CXCL2, CXCL5, CCL5, and CXCL8 were markedly decreased in the LPS-treated bovine retinal endothelial cells (BRECs) with 100 μg/ml of quercetin relatively to LPS alone. More importantly, the mRNA expressions of toll-like receptor 4 (TLR4), CD14, myeloid differential protein-2 (MD2), and myeloid differentiation primary response protein (MyD88) genes involved in TLR4 signal pathway were significantly attenuated by the addition of quercetin in LPS-treated BMEC, suggesting that quercetin can inhibit the TLR4 signal pathway. In addition, immunocytofluorescence showed that quercetin significantly inhibited the nuclear translocation of NF-κB p65 in BMEC induced by LPS. Therefore, the protective effects of quercetin on inflammatory responses in LPS-induced BMEC may be due to its ability to suppress the TLR4-mediated NF-κB signaling pathway. These findings suggest that quercetin can be used as an anti-inflammatory reagent to treat mastitis induced by exogenous or endogenous LPS release.

The purpose of the study was to assess the recovery, immune function, and breeding efficiency of postpartum dairy cows fed Astragalus membranaceus (AM) as a feed additive. The experiment used a completely randomized design. Cows were randomly assigned to two groups: (1) Control group fed total mixed ration (TMR; CON group, n = 15); (2) AM group fed TMR and AM (AM group, n = 15). The AM group was fed 675 g/day. The experimental results showed that compared with the CON group. The breeding interval of the AM group of dairy cows had a tendency to shorten (0.05 < p < 0.1). Plasma viscosity (PV), Plasma fibrinogen (FIB), the red cell aggregation index (TRCAI), Calcitonin (CT), Immunoglobulin M (IgM), and Luteinizing hormone (LH) results of AM group showed a time-treatment interaction (p < 0.05). Furthermore, the result of the study revealed that feeding AM as feed additives to dairy cows during the postpartum period had positive effects on wound recovery, immune function, endocrine regulation, and breeding efficiency.

The increased use of antibiotics continues to pose a threat to public health because of the increasing concern of antibiotic residue. Tea tree oil (TTO) is an extract of the Australian plant Melaleuca alternifolia with anti-inflammatory and antioxidant properties. However, there is little information on TTO supplementation in the diet of finishing pigs. Hence, the present study aimed to investigate the effect of TTO supplemented diets on the growth performance, meat quality, serum biochemical indices, and antioxidant capacity of the finishing pigs. Our results showed that TTO supplementation increased (P < 0.05) the mRNA expression of insulin-like growth factors -I (IGFs-I), growth acceleration hormone (GH), and heart fatty acid-binding protein (H-FABP), while the mRNA expression of myostatin gene (MSTN), and calpain-1 (CAST) decreased by the TTO supplementation, compared with the control group. In addition, TTO supplementation increased (P < 0.05) serum alkaline phosphatase (ALP), immunoglobulin G (IgG), and IgM levels but decreased (P < 0.05) serum aspartate transaminase (AST) concentration, relative to the control group. In addition, we found that the live weight and intramuscular fat enhanced (P < 0.05) significantly, and muscle pH 24 min value, cooking loss, and shear force decreased (P < 0.05) dramatically in the TTO group. The TTO supplementation increased (P < 0.05) C18:2n6t concentration and decreased (P < 0.05) C12:0 and C16:0 concentration, relative to the control group. Dietary supplementation with TTO decreased (P < 0.05) malondialdehyde (MDA) and increased (P < 0.05) glutathione peroxidase (GSH-Px) activity in serum. These results indicated that TTO supplementation could improve immunity and antioxidant, carcass traits, the nutritional value of pork, and the antioxidant capacity of finishing pigs. Therefore, TTO has potential positive effects as a feed additive in the pig industry.

This study introduces a methodology for detecting the location of signal sources within a metal plate using machine learning. In particular, the Back Propagation (BP) neural network is used. This uses the time of arrival of the first wave packets in the signal captured by the sensor to locate their source. Specifically, we divide the aluminum plate into several areas, design eight receiving points for receiving the excitation signal, and determine the location of each sound source. In order to train and test the machine learning network, the aluminum plate model was established using the COMSOL numerical simulation platform and the propagation of five peak waves was simulated. Correspondingly, experimental verification was carried out and a scanning laser Doppler vibrometer (SLDV) was used to build an experimental platform to collect the corresponding wave field information to obtain a data set for machine learning. The results show that the trained BP neural network can classify the sound source region in both environments.

This study aimed to evaluate whether sodium butyrate (SB) attenuates the ruminal response to LPS-stimulated inflammation by activating GPR41 in bovine rumen epithelial cells (BRECs). We examined the SB regulation of GPR41 and its impact on LPS-induced inflammation using GPR41 knockdown BRECs. The LPS-induced BRECs showed increases in the expression of genes related to pro-inflammation and decreases in the expression of genes related to tight junction proteins; these were attenuated by pretreatment with SB. Compared with that in LPS-stimulated BRECs, the ratio of phosphorylated NF-κB (p65 subunit) to NF-κB (p65 subunit) and the ratio of phosphorylated IκBα to IκBα were suppressed with SB pretreatment. The LSB group abated LPS-induced apoptosis and decreased the expression of Bax, Caspase 3, and Caspase 9 mRNA relative to the LPS group. In addition, the LSB group had a lower proportion of cells in the G0–G1 phase and a higher proportion of cells in the S phase than the LPS group. The mRNA expression of ACAT1 and BDH1 genes related to volatile fatty acid (VFA) metabolism were upregulated in the LSB group compared to those in LPS-induced BRECs. In addition, pretreatment with SB promoted the gene expression of GPR41 in the LPS-induced BRECs. Interestingly, SB pretreatment protected BRECs but not GPR41KD BRECs. Our results suggest that SB pretreatment protects against the changes in BRECs LPS-induced inflammatory response by activating GPR41.

The objective of this study was to determine the effect of replacing isonitrogenous and isoenergetic basis alfalfa hay (AH) with stevia (Stevia rebaudiana) hay in dairy cow diets on nutrient digestion, milk performance, rumen fermentation, and nitrogen (N) utilization. In this study, 24 healthy Holstein lactating dairy cattle with a similar milk yield of 33.70 ± 2.75 (mean ± SD) kg, days in milk 95.98 ± 23.59 (mean ± SD) days, and body weight 587.75 ± 66.97 (mean ± SD) kg were selected and randomly allocated into three groups. The constituents of the three treatments were (1) 30.0% AH, and 0% stevia hay (SH) for the AH group; (2) 24.0% AH, and 6% SH for the 6% SH group; (3) 18.0% AH, and 12% SH for the 12% SH group. The substitution of AH with SH did not affect dry matter intake (DMI), gross energy (GE), and other nutrients intake but increased the digestibility of neutral detergent fiber (NDF) and acid detergent fiber (ADF). Compared with the AH diet, the cows fed the 6% SH diet had a higher milk yield and concentration of milk fat. Fecal and urinary nitrogen (N) were lower in cows fed a 6% SH diet than in cows fed the AH diet. Milk N secretion and milk N as a percentage of N intake were higher in cows fed a 6% SH diet than in cows fed AH diets. The concentration of ruminal volatile fatty acids, acetic acid, and ammonia-N were higher in cows fed a 6% SH diet than in cows fed an AH diet. By comparison, the 12% SH group did not affect milk yield, milk composition, N utilization, and rumen fermentation compared with the AH and 6% SH groups. In conclusion, it appears that feeding 6% SH, replacing a portion of AH, may improve lactation performance and N utilization for lactating dairy cows.

The aim of this study was to evaluate the effects of buffalo milk and cow milk on lipid metabolism in obese mice. Milk composition analysis showed fat, protein, and total solid content in buffalo milk was higher than cow milk, while the lactose content of buffalo milk was lower than cow milk. After milk metabolite extraction and LC-MS/MS analysis, differential metabolites were mainly enriched in “linoleic acid metabolism pathways,” “pentose and glucuronate interconversion pathways,” and “metabolism of xenobiotics by cytochrome P450 pathways.” We fed three groups of C57BL/6J mice (n = 6 per group) for 5 weeks: (1) high-fat diet group (HFD group); (2) high-fat diet + buffalo milk group (HBM group); and (3) high-fat diet + cow milk group (HCM group). Our results showed that body weight of mice was significantly decreased in HBM and HCM groups from 1 to 4 weeks compared with the HFD group. The mRNA expression of ACAA2, ACACB, and SLC27A5 genes involved in the lipid metabolism in liver tissue were significantly elevated in HCM group, relatively to HFD and HBM group. In addition, the adipocyte number, size and lipid accumulation in the liver were significantly decreased in HCM group compared with the HFD group by H&E staining and oil red O staining, but was not change in HBM group. The mRNA levels of TNF-α and IL-1β inflammatory genes were significantly increased in HBM group, relatively to HFD and HCM group, which is consistent with results from inflammatory cell infiltration and tissue disruption by colon tissue sections. In conclusion, dietary supplementation of cow milk has beneficial effects on loss of weight and lipid metabolism in obese mice.

Abstract Background Tea tree oil (TTO) plays an important role in antibacterial activity and alleviating the inflammatory responses. Bovine mammary epithelium and polymorphonuclear leukocytes (PMNL) can actively respond to bovine mastitis infection. However, regulatory effects of TTO extracts on the innate immune response of bovine mammary epithelial cells (BMECs) and PMNL remain not reported. Therefore, aim of the study was to evaluate the effects of TTO extracts on the mRNA levels of the genes involved in the innate immune response of BMECs and PMNL. Results Our results demonstrated that addition of 0.025% and 0.05% TTO increased the proliferation of BMECs, and significantly enhanced (P

By means of tensile creep tests and microstructure analysis, the creep behavior and deformation mechanism of a nickel-based single-crystal alloy with small-angle deviations from the [111] orientation at 1040°C/137 MPa were investigated. The results suggested that in the early stage of creep, proliferating dislocations generated by different slip systems get accumulated in the γ phase. Dislocations mainly slipped in the γ phase and climbed over the γ′ phase, and they began to form a dislocation network at the interface. γ′ phases were connected to each other, and the coherence relationship was destroyed. In the steady-state stage of creep, the solid interfacial dislocation network was formed, and the γ′ phase transformed into a lamellar raft structure, which hinders the slip and climb of dislocations in the matrix channel. In the accelerated creep stage, the dislocation network was destroyed, and dislocations sheared into the γ′ phase in the way of dislocation pairs, which could react to form a superdislocation node and decompose to form APBs on the plane or cross slip from the plane to the plane to form K-W locks.

Tea tree oil (TTO) plays an important role in lipid metabolism, alleviating the inflammatory responses. Fatty liver is associated with lipid accumulation in hepatocytes, leading to inflammation. However, there is very limited information on the effects of TTO on lipid accumulation, and inflammation in bovine hepatocytes. This study aimed to evaluate whether TTO alleviates palmitic acid (PA)-induced lipid accumulation in bovine hepatocytes. Hepatocytes isolated from mid-lactating Holstein cows were pretreated with 100 μM PA for 72 h. Cells were either pretreated with PA alone (PA group) or with PA followed by 0.00625% TTO treatment for 12 h (PT group). Expression of fatty acid oxidant genes increased (P < 0.05) while fatty acid synthesis genes decreased (P < 0.05) in the PT group compared with the PA group. PA treatment resulted in increased (P < 0.05) expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), but these increases were less in the PT group (P < 0.05). Compared to the PA group, expression of phosphorylated (p)-p65 and p-inhibitor κBα (p-IκBα) was suppressed (P < 0.05) by TTO treatment. TTO treatment limited (P < 0.05) the increase in intracellular reactive oxygen species (ROS) and prevented (P < 0.05) a reduction in mitochondrial membrane potential observed in response to PA treatment. Expression of endoplasmic reticulum (ER) stress genes was reduced (P < 0.05) in the PT group compared with the PA group. Our results suggest that TTO treatment attenuates the effects of PA in hepatocytes, leading to fatty acid oxidation, decreased fatty acid synthesis, suppressed inflammatory response, and reduced ER stress. Taken together, the results of this study suggest that TTO treatment may be a promising therapeutic approach to imbalanced lipid homeostasis, inflammation and ER stress in dairy cows shortly before and after calving.

Tea tree oil (TTO) plays an important role in regulating lipid metabolism and has anti-inflammatory properties. In postpartum dairy cows, dry matter intake (DMI) is dramatically decreased, resulting in lipid metabolism disorder and the systemic pro-inflammatory response. However, the effects of TTO on glucolipid metabolism and immunity in postpartum dairy cows remain uninvestigated. Therefore, this study aimed to evaluate the effects of TTO on production performance, serum biochemical indicators, and immunity in postpartum dairy cows. Our results demonstrate that DMI tended to increase (p = 0.07) in the total mixed ration (TMR) diets supplemented with 0.01% TTO/dry matter (DM) basis relative to that in the control group. The 4% fat-corrected milk (FCM) content in the 0.01% and 0.02% TTO groups showed an increase (p = 0.09) compared with that in the control. Remarkably, the levels of globulin (GLO) and immunoglobulin G (IgG) were elevated (p < 0.05) in the TMR diet supplemented with 0.02% TTO compared to those in the control group. The TTO caused no profound changes in cholesterol (CHO), triglyceride (TG), high-density lipoprotein (HDL), or low-density lipoprotein (LDL). Notably, 0.02% TTO increased (p < 0.05) the serum glucose concentration relative to that in the control group. In conclusion, our results demonstrate that TTO could improve glucolipid metabolism and enhance immunity in postpartum dairy cows. It may be a novel resolution strategy for body condition recovery and the improvement of milk performance.

Fermentation of agricultural by-products by white rot fungi is a research hotspot in the development of ruminant feed resources. The aim of this study was to investigate the potential of the nutritional value and rumen fermentation properties of white tea residue fermented at different times, using single and dual culture white rot fungal species. Phanerochaete chrysosporium, Pleurotus ostreatus, and Phanerochaete chrysosporium + Pleurotus ostreatus (dual culture) solid-state fermented white tea residue was used for 4 weeks, respectively. The crude protein content increased significantly in all treatment groups after 4 weeks. Total extractable tannin content was significantly decreased in all treatment groups (p < 0.01). P. chrysosporium and dual culture significantly reduced lignin content at 1 week. The content of NH3-N increased in each treatment group (p < 0.05). P. chrysosporium treatment can reduce the ratio of acetic to propionic and improve digestibility. Solid state fermentation of white tea residue for 1 week using P. chrysosporium was the most desirable.

Cellulase enzymes attract a lot of research due to their industrial application. Diverse cellulase-producing organisms and substances that induce cellulase are highly sought after. This study aimed to evaluate the effect of different inducer sources on cellulase production by white rot fungi P. ostreatus CGMCC 3.7292 and P. chrysosporium CGMCC 3.7212 under submerged fermentation employing a completely randomized experimental design. The different inducer sources tested were nitrogen (yeast, potassium nitrate, sodium nitrate, ammonium sulphate, aqueous ammonia and urea), carbon (malt extract, glucose, fructose, carboxymethylcellulose, starch and xylose) and agro-biomass (stevia straw, wheat straw, oat straw, alfalfa straw, corn cobs and corn stover). These inducer sources strongly impacted enzyme activities by P. ostreatus CGMCC 3.7292 and P. chrysosporium CGMCC 3.7212. The suitable nitrogen and carbon inducer sources for cellulase activity by P. ostreatus and P. chrysosporium were yeast (1.354 U/mL and 1.154 U/mL) and carboxymethylcellulose (0.976 U/mL and 0.776 U/mL) while the suitable agro-biomass were wheat straw (6.880 U/mL) and corn stover (6.525 U/mL), respectively. The least inducer sources in terms of nitrogen, carbon and agro-biomass for cellulase activity by P. ostreatus and P. chrysosporium were urea (0.213 U/mL and 0.081 U/mL), glucose (0.042 U/mL and 0.035), xylose (0.042 U/mL and 0.035 U/mL) and stevia straw (1.555 U/mL and 0.960 U/mL). In submerged fermentation, the cellulase enzyme activity of P. ostreatus in response to various inducer sources was relatively higher than P. chrysosporium.

Acetate is a precursor substance for fatty acid synthesis in bovine mammary epithelial cells (BMECs), and the mTOR signaling pathway plays an important role in milk fat synthesis. However, the mechanism of the regulatory effects of acetate on lipogenic genes via the mTOR signaling pathway in BMEC remains unknown. We hypothesized that acetate can enhance the expression of lipogenic genes and triglyceride (TG) production by activating the mTOR signaling pathway in BMECs. Therefore, the aim of this study was to investigate the network of acetate-regulated lipid metabolism by the mTOR signaling pathway in BMECs. These results showed that TG synthesis was elevated (p < 0.01) in BMECs with acetate treatment. The lipid droplets were increased in the acetate-treated groups compared with those in the control group through the Bodipy staining of the lipids. In addition, the fatty acid profile in BMECs treated with acetate was affected, with an elevation in the proportions of C14:0, C16:0, and C18:0. The mRNA levels of the sterol-response-element-binding protein 1 (SREBP1), stearoyl-CoA desaturase 1 (SCD1), and fatty acid synthase (FAS) genes involved in the lipogenesis and transcriptional factors were upregulated (p < 0.05) in BMECs with acetate treatment. Remarkably, the expression of acetyl-CoA carboxylase α (ACCα) and FAS rate-limiting enzymes involved in lipogenesis was upregulated in BMECs with acetate treatment. Moreover, the addition of acetate enhanced the key protein expression of S6K1, which is related to the mTOR signaling pathway. Taken together, our data suggest that TG accumulation and expression of lipogenic genes induced by acetate are associated with the activation of the mTOR signaling pathway, which provides new insights into the understanding of the molecular mechanism in the expression of mTOR-signaling-pathway-regulated lipogenic genes.

The E2F family of transcription factor is divided into activators and repressors that control cell proliferation. Bovine mammary epithelial cells (BMECs) can be immortalized using human papillomavirus 16 E6E7 (HPV16 E6E7) and simian vacuolating virus 40 large T antigen (SV40T). In addition, SV40T does not require E2F1, E2F2, and E2F3 activators to induce proliferation in mouse embryo fibroblasts (MEFs). However, we report that E2F3 activator is required to induce the proliferation of BMECs. Our results showed that, at an early stage, primary BMECs lacking the E2F1 expression have the capacity to proliferate and show E2F2 and E2F3 slight protein levels. At a late stage, primary BMECs deficient for E2F3 completely abolish any proliferative ability and exhibit a severe cell senescence signal, although the E2F2 can be expressed at a late stage of primary BMECs. Compared with the late stage of primary BMECs, the BMECs immortalized by SV40T and E6E7 restored the protein level of E2F3 and enhanced the CDK4, CDK6, cyclin D3, and CDK2 protein level, leading to proliferating robustly. Surprisingly, it was found that p53, p21Cip1, and p27Kip1 were upregulated in SV40T and E6E7-immortalized BMECs, relatively to primary BMECs. Notably, Cdc2 was almost expressed in primary BMECs. However, Cdc2 was elevated in BMECs immortalized by SV40T and E6E7. In conclusion, this study revealed a molecular mechanism where E2F3 controls the BMECs’ proliferation and senescence.

In the context of global restrictions on the use of antibiotics, there has been increased research on natural plant-based ingredients as additives. It has been proved that many natural active ingredients contained in plants have positive effects on animal growth regulation. Artemisia argyi (A. argyi) is a traditional Chinese herbal medicine, and its extracts have been reported to have a variety of biological activities. Therefore, in order to explore the potential of the active extract of Artemisia argyi leaves (ALE) as a plant source additive, mice were fed with ALE at different concentrations for 60 days. Finally, the effects of ALE were evaluated by the growth indexes, blood indexes, and intestinal microflora changes of the mice. It was found that a medium concentration of ALE (150 mg/kg) could promote growth, and especially improved the feed efficiency of the mice. However, high concentrations of ALE (300 mg/kg) had some negative effects on the growth of mice, especially liver damage, which significantly increased AST and ALT levels in the blood. Therefore, the 150 mg/kg ALE treatment group was selected for 16S rDNA analysis. It was found that ALE could play a positive role by regulating the proportion of Bacteroidetes and Firmicutes in the intestinal tract. In particular, it can significantly up-regulate the quantities of Akkermansia and Bifidobacterium. These results suggest that ALE at appropriate concentrations can positively regulate animal growth.

In this work, two types of self-healing composite laminates with embedded 6 and 12 wt% microcapsules respectively, were prepared. The guided waves were activated and captured using piezoelectric transducer in a pitch-catch approach. Mode I interlaminar fracture tests were conducted using double cantilever beam specimens, and the effects of concentration of microcapsules on the interlaminar properties of specimens were investigated. The interlaminar fracture surfaces of self-healing specimens were observed to support the analysis of mechanical tests and guided wave detecting results. The results show that the self-healing efficiency of specimens with high-concentration microcapsules is less than that of specimens with low-concentration microcapsules. Meanwhile, low-concentration microcapsules reduce the interlaminar properties of specimens. Two guided-wave parameters, namely signals peak value and waveform similarity, were set and correlated with the self-healing capability of specimens. When the specimens were damaged, the peak value of response signals decreases, and the waveform difference between the response signal and reference signal becomes larger. When the specimens were healed, the peak value of response signals rises, and the waveform difference between them becomes smaller.

Various pretreatments are employed to increase the utilization of rice straw as a ruminant feed ingredient to minimize its negative environmental impact. However, an efficient alternative is still needed. The purpose of this study was to evaluate the ability of ammonia and/or white-rot fungi (Pleurotus ostreatus) to degrade lignin, increase the nutritional value, and enhance the rumen fermentability of rice straw. Rice straw was treated with ammonia and/or basidiomycete white-rot fungi (P. ostreatus) with untreated straw as control under solid-state fermentation employing a completely randomized design. The crude protein increased from 2.05% in the control to 3.47% in ammoniated rice straw, 5.24% in basidiomycete white-rot fungi (P. ostreatus), and 6.58% in ammoniated-basidiomycete white-rot fungi-treated (P. ostreatus) rice straw. The ammoniated-basidiomycete white-rot fungi-treated (P. ostreatus) rice straw had the least lignin content (3.76%). Ammoniated-basidiomycete white-rot fungi-treated (P. ostreatus) rice straw had improved in vitro dry matter digestibility (65.52%), total volatile fatty acid (76.56 mM), and total gas production (56.78 mL/g) compared to ammoniated rice straw (56.16%, 67.71 mM, 44.30 mL/g) or basidiomycete white-rot fungi-treated (P. ostreatus) rice straw (61.12%, 75.36 mM, 49.31 mL/g), respectively. The ammoniated-basidiomycete white-rot fungi (P. ostreatus) treatment improved rice straw’s nutritional value, in vitro dry matter digestibility, volatile fatty acids, and gas production.

Lipopolysaccharide (LPS) is an endotoxin that induces immune and inflammatory responses in the rumen epithelium of dairy cows. It is well-known that flavonoid phloretin (PT) exhibits anti-oxidative, anti-inflammatory and antibacterial activity. The aim of this research was to explore whether PT could decrease LPS-induced damage to bovine rumen epithelial cells (BRECs) and its molecular mechanisms of potential protective efficacy. BRECs were pretreated with PT for 2 h and then stimulated with LPS for the assessment of various response indicators. The results showed that 100 µM PT had no significant effect on the viability of 10 µg/mL LPS-induced BRECs, and this dose was used in follow-up studies. The results showed that PT pre-relieved the decline in LPS-induced antioxidant indicators (T-AOC and GSH-PX). PT pretreatment resulted in decreased interleukin-1β (IL-1β), IL-6, IL-8, tumor necrosis factor-α (TNF-α) and chemokines (CCL2, CCL5, CCL20) expression. The underlying mechanisms explored reveal that PT may contribute to inflammatory responses by regulating Toll-like receptor 4 (TLR4), nuclear transcription factor-κB p65 (NF-κB p65), and ERK1/2 (p42/44) signaling pathways. Moreover, further studies found that LPS-induced BRECs showed decreased expression of claudin-related genes (ZO-1, Occludin); these were attenuated by pretreatment with PT. These results suggest that PT enhances the antioxidant properties of BRECs during inflammation, reduces gene expression of pro-inflammatory cytokines and chemokines, and enhances barrier function. Overall, the results suggest that PT (at least in vitro) offers some protective effect against LPS-induced ruminal epithelial inflammation. Further in vivo studies should be conducted to identify strategies for the prevention and amelioration of short acute rumen acidosis (SARA) in dairy cows using PT.

Twenty-four healthy castrated male Holstein growing cattle, with similar body weight (301 ± 11.5 kg), were enrolled in this study and randomly allocated into two groups (12/pen). Holstein growing cattle in the LPT (low NFC/NDF pelleted TMR) group were fed basal pelleted TMR with a low NFC/NDF ratio (NFC/NDF = 1.07), while the HPT (high NFC/NDF pelleted TMR) group were fed with a high NFC/NDF ratio diet (NFC/NDF = 1.71). The results showed that: (1) Body measurements were found to be increased for the LPT group (p < 0.05); compared with the HPT group, feed intake to gain ratio and feed cost in the LPT group were decreased by 12.24% and 15.35%, respectively (p < 0.01). Compared with the HPT group, the LPT group tended to increase chest girth. (2) Digestibility of DM and NDF in the LPT group was higher (p < 0.05) than in the HPT group, being increased by 3.41% and 4.26%, respectively, and increased digestibility of ADF in the LPT group was significant (p < 0.01). (3) The daily feed consumption of NDF in the LPT group was higher than that in the HPT group and the daily rumination time and chewing time in the LPT group were longer than that in the HPT group (p < 0.05). (4) Compared with the LPT group, the parameter of pH, microbial protein and acetate: propionate (p < 0.05) in the HPT group were decreased by 8.57%, 12.46% and 23.71%, respectively. In contrast, the concentration of total volatile fatty acids, acetate and propionate were higher (p < 0.05) in the HPT group, and increased by 13.49%, 19.59% and 52.70%, respectively. (5) Compared with the LPT group, rumen fluid in the HPT group diet up-regulated the mRNA expression levels of BRECs pro-inflammatory factor IL-1β and TNF-α (p < 0.05), and meanwhile, up-regulated the mRNA expression levels of BRECs pro-inflammatory factor IL-6 (p < 0.01); compared with the LPT group, rumen fluid in the HPT group diet up-regulated the mRNA expression levels of CCL28 and CCL20 (p < 0.05) chemokines in CCL types of BRECs; in addition, compared with the LPT group, rumen fluid in the HPT group up-regulated the mRNA expression levels of CXCL2, CXCL3, CXCL9 and CXCL14 chemokines in CXCL types of BRECs (p < 0.01), and the mRNA expression levels of the CXCL5 chemokine tended to be increased (p = 0.06).

In the context of the unsatisfactory therapeutic effect of antibiotics, the natural products of plants have become a research hotspot. Artemisia argyi (A. argyi) is known as a traditional medicine in China, and its extracts have been reported to have a variety of active functions, including anti-inflammatory. Therefore, after establishing the mouse mastitis model by lipopolysaccharide (LPS), the effects of A. argyi leaves extract (ALE) were evaluated by pathological morphology of the mammary gland tissue, gene expression, and serum oxidation index. Studies have shown that ALE has a restorative effect on LPS-induced mammary gland lesions and significantly down-regulated the rise of myeloperoxidase (MPO) induced by LPS stimulation. In addition, ALE played a positive role in LPS-induced oxidative imbalance by restoring the activities of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) and preventing the increase in nitric oxide (NO) concentration caused by the over-activation of total nitric oxide synthase (T-NOS). Further analysis of gene expression in the mammary gland showed that ALE significantly down-regulated LPS-induced up-regulation of inflammatory factors IL6, TNFα, and IL1β. ALE also regulated the expression of MyD88, a key gene for toll-like receptors (TLRs) signaling, which, in turn, regulated TLR2 and TLR4. The effect of ALE on iNOS expression was similar to the effect of T-NOS activity and NO content, which also played a positive role. The IκB gene is closely related to the NF-κB signaling pathway, and ALE was found to significantly alleviate the LPS-induced increase in IκB. All of these results indicated that ALE may be considered a potential active substance for mastitis.

Naturally occurring plant compounds including tannins, saponins and essential oils are extensively assessed as natural alternatives to in-feed antibiotics. Tannins are a group of polyphenolic compounds that are widely present in plant region and possess various biological activities including antimicrobial, anti-parasitic, anti-viral, antioxidant, anti-inflammatory, immunomodulation, etc. Therefore, tannins are the major research subject in developing natural alternative to in-feed antibiotics. Strong protein affinity is the well-recognized property of plant tannins, which has successfully been applied to ruminant nutrition to decrease protein degradation in the rumen, and thereby improve protein utilization and animal production efficiency. Incorporations of tannin-containing forage in ruminant diets to control animal pasture bloat, intestinal parasite and pathogenic bacteria load are another 3 important applications of tannins in ruminant animals. Tannins have traditionally been regarded as “anti-nutritional factor” for monogastric animals and poultry, but recent researches have revealed some of them, when applied in appropriate manner, improved intestinal microbial ecosystem, enhanced gut health and hence increased productive performance. The applicability of plant tannins as an alternative to in-feed antibiotics depends on many factors that contribute to the great variability in their observed efficacies. Keywords: Tannins, Biological activities, In-feed antibiotics, Natural feed additives, Farm animal

Histidine (His) is considered to be the first-limiting amino acid (AA) on grass silage-based diets in lactation cows, which correlate positively with lactose yield. The higher glucose requirements of lactating cows can be met through a combination of increased capacity for gluconeogenesis and increased supply of gluconeogenic precursors. However, the effect of His on the expression of gluconeogenic genes in the bovine hepatocytes is less known. Therefore, this study aimed to investigate the regulatory effect of His on the key gluconeogenic genes and glucose output in bovine hepatocytes. The addition of 0.15, 0.6, and 1.2 mM His in a medium significantly enhanced (p < 0.05) the viability of bovine hepatocytes. Remarkably, 1.2 mM His induced profound changes (p < 0.05) in the mRNA level of key genes involved in gluconeogenesis, including PCK1, PCK2, FBP1, and G6PC in vitro. Furthermore, the mRNA expression of PCK1 was significantly elevated (p < 0.05) by the addition of 1.2 mM His at 3, 6, 12, and 24 h of incubation. The hepatic glucose output increased (p < 0.05) linearly with increasing His concentration. These findings indicate that the addition of His may be efficiently converted into glucose via the upregulation of genes related to the gluconeogenic pathway.

Sixty castrated male Haimen white growing goats with an initial age of 100 days old and similar body weight (16 ± 1.5 kg) were selected and randomly allocated into five groups with three replicates in each group with four goats in each pen (5 m × 3.2 m). Goats in the control group (CG) were fed a basal pelleted total mixed ration supplemented with 30% alfalfa hay pellet, while experimental treatments (G6, G12, G18, or G24) were supplemented with four levels (6%, 12%, 18%, or 24%) of GBLR replacing alfalfa hay pellet in the diet, separately. Results showed that (1) the final body weight, average daily gain, and average feed intake of G18 was significantly higher (p < 0.05) than CG; in contrast, the feed conversion ratio of G18 was significantly lower than CG and G12 (p < 0.05); the feed cost per head per day of CG was significantly higher (p < 0.05) than that of G18 and G24, and exhibited obvious linearly decrease (p = 0.04) with increasing GBLR supplementation; and apparent total-tract digestibility of DM and NDF in GBLR treatments were significantly higher (p < 0.05) than CG; (2) alanine transaminase (ALT) concentration in the G18 group was significantly lower (p < 0.05) than those in the control group; aspartate transaminase (AST) concentration in the G24 was significantly lower (p < 0.05) than those in the control group, and an increase in dietary level of GBLR tended to result in a linear decrease (p = 0.09) in the concentration of serum AST; (3) the concentration of malondialdehyde (MDA) demonstrated a tendency to decrease (p = 0.06) linearly with increasing GBLR supplementation; however, glutathione peroxidase (GSH-PX) activity in G12 was significantly higher (p < 0.05) than CG, G6, and G24; in addition, superoxide dismutase (SOD) activity in G18 was significantly higher (p < 0.05) than CG and G6; concentration of immunoglobulin M (IgM), immunoglobulin G (IgG), and immunoglobulin A (IgA) were not affected by GBLR, but increasing dietary GBLR showed a tendency (p = 0.08) to linearly increase the IgG concentration; the content of interleukin 4 (IL4) was significantly higher (p < 0.05) in G12, G18, and G24 than that in CG and G6; (4) There were similar NH3-N, pH, TVFA, and butyrate for goats fed different levels of GBLR supplementation; the C2 (p = 0.07) and acetate: propionate (p = 0.06) demonstrated a tendency to increase linearly with increasing level of GBLR supplementation, separately; however, it was observed that concentration of propionate showed a tendency to decrease (p = 0.08) linearly in response to GBLR supplementation; and (5) Increasing dietary GBLR tended to linearly enhance the lightness (L*) (p = 0.07) and yellowness (b*) (p = 0.09) values of longissimus dorsi muscles; the redness (a*) value in G18 was significantly higher than that in CG (p < 0.05).

Bacillus megaterium is an ideal microecologics in the feed industry. BM1259 was already isolated from chicken manure and the whole-genome sequencing was also analyzed in our previous study. However, few studies concentrated on dietary supplementation with BM1259 in young ruminants and especially its effect on Holstein bull calves have not been reported. Hence, this experiment was conducted with the aim to evaluate the effects of BM1259 on growth performance, nutrient digestibility, rumen fermentation, and blood biochemical parameters in Holstein bull calves. Twenty-four healthy Holstein bull calves with the initial age of 90 days old and a similar body weight (115 ± 6.5 kg) were selected and randomly allocated into two groups with one Holstein bull calf in each pen (2.5 m × 2.2 m). Holstein bull calves in the control group (COG) were fed a basal total mixed ration (TMR), while experimental treatments (BMG) were fed with the TMR diet supplemented with 12 g/head/day of BM1259 powder (1 × 1010 cfu/g) separately. Results showed that (1) the average daily gain and dry matter intake of the BMG were significantly higher than those of the COG (p < 0.01), increased by 12.5% and 8.79%, respectively, during the 4–8 weeks after the addition of 12 g/head/day of BM1259; from 0 to 8 weeks, ADG (p < 0.05) and DMI (p < 0.05) in the BMG were significantly higher than those in the COG, increased by 14.9% and 6.04%, respectively. (2) At the end of the fourth week, the apparent digestibility of crude protein and neutral detergent fiber in the BMG was significantly higher than that in the COG (p < 0.05), increased by 5.97% and 6.70%, respectively; at the end of the eighth week, the apparent digestibility of crude protein and neutral detergent fiber was significantly higher than that of the COG (p < 0.01), increased by 5.88% and 10.26%, respectively. (3) At the end of the eighth week, the rumen fluid pH (p < 0.05), MCP (p < 0.05), and acetate (p < 0.05) in the BMG were significantly higher than those in the COG, increased by 9.03%, 19.68%, and 12.74%, respectively; at the end of the fourth and eighth week, NH3-N concentration in the BMG was significantly lower than that in the COG, with a decrease of 21.81% and 16.40%, respectively. (4) At the end of the fourth (p < 0.05) and eighth week (p < 0.05), the glutamate content of the rumen fluid of the Holstein bull calves in the BMG was significantly higher than that in the COG, increased by 13.21% and 14.32%, respectively; at the end of the fourth week, the contents of glutamate in the serum (p < 0.05), urine (p < 0.05), and feces (p < 0.05) of the Holstein bull calves in the BMG were significantly lower than those in the COG, decreased by 25.76%, 33.87%, and 9.23%, respectively; at the end of the eighth week, the contents of glutamate in the serum, urine, and feces of the Holstein bull calves in the BMG were significantly lower than those in the COG (p < 0.01), decreased by 26.69%, 27.94%, and 11.11%, respectively. (5) After adding 12 g/head/day of BM1259, the urine ammonia–nitrogen content of the BMG was extremely significantly lower than that of the COG at the end of the fourth and eighth week (p < 0.01), decreased by 54.60% and 40.31%, respectively. (6) After adding 12 g/head/day of BM1259, there was no significant effect on the level of blood biochemical parameters of the Holstein bull calves. This study demonstrates that BM1259 can be applied as a potential microecologics to improve growth performance, nutrient digestibility, rumen fermentation, and nitrogen utilization in Holstein bull calves.

In subacute rumen acidosis (SARA), the rumen epithelium is frequently attacked by endotoxin (LPS), which is caused by the lysis of dead Gram-negative bacteria. However, the rumen epithelium innate immune system can actively respond to the infection. Previous studies have demonstrated that tea tree oil (TTO) has good bactericidal and anti-inflammatory effects. Therefore, the aim of this study was to investigate the effect of TTO on the expression of genes involved in the inflammatory cytokines in goat rumen epithelial cells (GRECs) triggered by LPS. Our study shows that rumen epithelial cells isolated from goat rumen tissue can be cultured in vitro in 0.25% trypsin for a long time. These cells were identified as epithelial cells by the expression of cytokeratin 18, monocarboxylate transporter 4 (MCT4), Na[+]/H[+] hydrogen exchanger 1 (NHE1), putative anion transporter 1 (PAT1), vH+ ATPase B subunit (vH+ ATPase), and anion exchanger 2 (AE2). The mRNA expression of IL-1β, IL-6, TNF-α, TLR-2, NF-κB, CXCL6 and CXCL8 genes was significantly increased when LPS was used compared to untreated controls. In addition, mRNA expression of IL-1β, IL-6, TNF-α, TLR-2, NF-κB, CXCL8, CXCL6 and interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) genes was also significantly higher in the LPS group compared to the 0.05% TTO group. However, the expression of IL-1β, IL-6, TNF-α, TLR-2, CXCL6 and IFIT3 genes was significantly lower in the LPS and 0.05% TTO group compared to the 1 μg/mL LPS group. These results suggest that TTO can inhibit LPS-induced inflammatory cytokines expression in GRECs.

Through creep performance tests, microstructural observations, and contrast analysis of the dislocation configuration, the deformation and damage mechanism of the directionally solidified nickel-based superalloy during creep at moderate temperatures was investigated. The findings suggested that the deformation of the alloy in the late stage of creep at moderate temperatures involved dislocations slipping in the γ matrix and shearing into the γ′ phase. The super-dislocations sheared into the γ′ phase could either be decomposed to form a super-Shockley incomplete dislocation plus superlattice intrinsic stacking fault (SISF) configuration, or it could slip from the {111} plane to the {100} plane and decompose to form a dislocation configuration of the Kear–Wilsdorf (K-W) lock plus antiphase domain boundary (APB). The configurations of the dislocations could inhibit the slipping and cross-slipping of dislocations to enhance the alloy creep strength, which is thought to be one reason that the alloy displayed good creep resistance. In the late creep stage, the primary/secondary slipping systems were alternately activated, and the interaction of the slipping traces caused micro-holes to appear on the interface of the γ/γ′ phases at the intersection areas of the two slipping systems. The micro-holes gathered and grew to form micro-cracks, which extended along the grain boundary at 45° to the stress axis until creep rupture occurred. These were the damage and fracture characteristics of the alloy in the late stage of creep at moderate temperatures.

Objective The objective of this study was to examine the effects of alfalfa flavonoids on the production performance, immunity, and ruminal fermentation of dairy cows. Methods The experiments employed four primiparous Holstein cows fitted with ruminal cannulas, and used a 4×4 Latin square design. Cattle were fed total mixed ration supplemented with 0 (control group, Con), 20, 60, or 100 mg of alfalfa flavonoids extract (AFE) per kg of dairy cow body weight (BW). Results The feed intake of the group receiving 60 mg/kg BW of AFE were significantly higher (p

This experiment was conducted to study the effects of dietary alfalfa flavonoids extraction supplemental level on growth performance, organ development and blood biochemical indexes of Yangzhou geese at the age of 28 to 70 days. Two hundred and forty 21-day-old healthy male geese with similar body weight were randomly distributed into 4 groups with 6 replicates per group and 10 geese per replicate. Geese in the control group were fed a basal diet and the others in the experimental groups (groups 1, 2, and 3) were fed experimental diets supplemented with 150, 300 and 450 mg/kg alfalfa flavonoids extraction (the concentration of it was 81%), respectively. The experiment had 7 days for pre-test and 42 days for formal test. The results showed that the final body weight and average daily intake of group 2 were significantly higher than those of other groups (P 0.05). Pre-slaughter live weight, carcass weight, slaughter rate, semi-eviscerated weight, semi-eviscerated rate, eviscerated weight, eviscerated rate, leg muscle weight and leg muscle rate had no significant difference between each group (P > 0.05). The breast muscle weight and ratio of each test group were significantly higher than those in the control group (P 0.05). Spleen weight in test groups was significantly higher than that in the control group (P 0.05). Alanine aminotransferase and aspartate transaminase (AST) in groups 2 and 3 were higher than those in the group 1 and control group but not obvious (P > 0.05) and alkaline phosphatase (ALP) in groups 1 and 2 was higher than that in the control group and group 3 (P > 0.05). It is concluded that alfalfa flavonoids extraction added in dietary feed improve the growth performance, organ development and blood biochemical indexes of Yangzhou geese. It is concluded that 300 mg/kg supplemental level of the dietary alfalfa flavonoids extraction is optimal in this experiment.

The rumen immune system often suffers when challenging antigens from lysis of dead microbiota cells in the rumen. However, the rumen epithelium innate immune system can actively respond to the infection. Previous studies have demonstrated G protein-coupled receptors 41 (GPR41) as receptors for short chain fatty acids (SCFAs) in human. We hypothesized that SCFAs, the most abundant microbial metabolites in rumen, may regulate the immune responses by GPR41 in bovine rumen epithelial cells (BRECs). Therefore, the objective of study was to firstly establish an immortal BRECs line and investigate the regulatory effects of SCFAs and GPR41 on innate immunity responses in BRECs. These results showed that long-term BRECs cultures were established by SV40T-induced immortalization. The concentrations of 20 mM SCFAs significantly enhanced the levels of GPR41, IL1β, TNFα, chemokines, and immune barrier genes by transcriptome analysis. Consistent with transcriptome results, the expression of GPR41, IL1β, TNFα, and chemokines were markedly upregulated in BRECs treated with 20 mM SCFAs by qRT-PCR compared with control BRECs. Remarkably, the GPR41 knockdown (GPR41KD) BRECs treated with 20 mM SCFAs significantly enhanced the proinflammatory cytokines IL1β and TNFα expression compared with wild type BRECs treated with 20 mM SCFAs, but reduced the expression of CCL20, CXCL2, CXCL3, CXCL5, CXCL8, CXCL14, Occludin, and ZO-1. Moreover, GPR41 mRNA expression is positively correlated with CCL20, CXCL2, CXCL3, CXCL8, CXCL14, and ZO-1. These findings revealed that SCFAs regulate GPR41-mediated levels of genes involved in immune cell recruitment and epithelial immune barrier and thereby mediate protective innate immunity in BRECs.

In this study, we isolated a novel bacterium, Bacillus megaterium 1259 (BM1259), from chicken manure. Whole-genome sequencing analysis showed that the BM1259 complete genome is composed of a 5,043,095 bp circular chromosome and three circular plasmids, and it encodes 5379 coding genes and 182 RNA genes. Among these genes, a series of nitrate assimilation-related genes and pathways were identified, implying a potential role of BM1259 in nitrate metabolism. In addition, 24 lactating Holstein dairy cows were randomly assigned to four groups that were fed a total mixed ration (TMR) diet only (C), a TMR diet supplemented with 5 g/day of BM1259 (T1), a TMR diet supplemented with 10 g/day of BM1259 (T2), or a TMR diet supplemented with 15 g/day of BM1259 (T3). The results showed that supplementing dairy cows with 15 g/day of BM1259 increased 4% fat-corrected milk production. The molar proportion of propionate (C3) was significantly higher in T2 than in C. The C2:C3 ratio of T3 was higher than those of C and T2. No negative effect of BM1259 on blood indicators was detected. This study demonstrates BM1259 can be applied as a potential probiotic to improve nitrogen utilization and milk production in lactating dairy cows.

In recent years, 3D printing has received increasing attention from researchers. This technology overcomes the limitations of traditional technologies by printing precise and personalized scaffold with arbitrary shapes, pore structures, and porosities for the applications in various tissues. The cellulose nanocrystal (CNC) is extracted from Humulus Japonicus (HJS) and mixed with poly(ε-caprolactone) (PCL) to prepare a series of CNC/PCL composites for printing. Based on the analysis of the physical and chemical properties of the series of the CNC/PCL composites, an optimal mass ratio of CNC to PCL was obtained. The Solidworks was used to simulate the stretching and compression process of the scaffolds with three different patterns under an external force. The flow of nutrient solution in the scaffolds with different patterns was simulated by ANSYS FLUENT, and then a new optimization scaffold pattern with a concave hexagon shape was advised based on the simulation results. Collectively, the mechanical test results of the material and scaffold confirmed that the optimal filling amount of the CNC was 5%, and the scaffold pattern with concave hexagon shape exhibited better mechanical properties and suitable for the transport of cells and nutrients, which is expected to be more widely used in 3D printing.