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The dietary rumen-degradable starch (RDS) to rumen-degradable protein (RDP) ratio, denoted as the RDS-to-RDP ratio (SPR), has been proven to enhance in vitro rumen fermentation. However, the effects of dietary SPR in vivo remain largely unexplored. This study was conducted to investigate the effect of dietary SPR on lactation performance, nutrient digestibility, rumen fermentation patterns, blood indicators, and nitrogen (N) partitioning in mid-lactating Holstein cows. Seventy-two Holstein dairy cows were randomly assigned to three groups (24 head/group), balanced for (mean ± standard deviation) days in milk (116 ± 21.5), parity (2.1 ± 0.8), milk production (42 ± 2.1 kg/d), and body weight (705 ± 52.5 kg). The cows were fed diets with low (2.1, control), medium (2.3), or high (2.5) SPR, formulated to be isoenergetic, isonitrogenous, and iso-starch. The study consisted of a one-week adaptation phase followed by an eight-week experimental period. The results indicated that the high SPR group had a lower dry matter intake compared to the other groups (p

The DAEase gene sequence derived from Clostridium bolteae ATCCBAA-613 was synthesized by codon optimization. Using pCold TF as the expression vector, the cold-shock promoter CspA induced the expression of the DAEase gene in Escherichia coli BL21(DE3) at low temperature. Then, the highly soluble recombinant Cb-DAEase was obtained and purified by Ni-chelating affinity chromatography. Results showed that, the Cb-DAEase exhibited maximum activity at pH7.0 and 55 ℃. Additionally, the Cb-DAEase showed different sensitivities to the various metal ions when Co2+ was able to significantly (P

Interpersonal communication through vocal information is very important for human society. During verbal interactions, our vocal cord vibrations convey important information regarding voice identity, which allows us to decide how to respond to speakers (e.g., neither greeting a stranger too warmly or speaking too coldly to a friend). Numerous neural studies have shown that identifying familiar and unfamiliar voices may rely on different neural bases. However, the mechanism underlying voice identification of individuals of varying familiarity has not been determined due to vague definitions, confusion of terms, and differences in task design. To address this issue, the present study first categorized three kinds of voice identity processing (perception, recognition and identification) from speakers with different degrees of familiarity. We defined voice identity perception as passively listening to a voice or determining if the voice was human, voice identity recognition as determining if the sound heard was acoustically familiar, and voice identity identification as ascertaining whether a voice is associated with a name or face. Of these, voice identity perception involves processing unfamiliar voices, and voice identity recognition and identification involves processing familiar voices. According to these three definitions, we performed activation likelihood estimation (ALE) on 32 studies and revealed different brain mechanisms underlying processing of unfamiliar and familiar voice identities. The results were as follows: (1) familiar voice recognition/identification was supported by a network involving most regions in the temporal lobe, some regions in the frontal lobe, subcortical structures and regions around the marginal lobes; (2) the bilateral superior temporal gyrus was recruited for voice identity perception of an unfamiliar voice; (3) voice identity recognition/identification of familiar voices was more likely to activate the right frontal lobe than voice identity perception of unfamiliar voices, while voice identity perception of an unfamiliar voice was more likely to activate the bilateral temporal lobe and left frontal lobe; and (4) the bilateral superior temporal gyrus served as a shared neural basis of unfamiliar voice identity perception and familiar voice identity recognition/identification. In general, the results of the current study address gaps in the literature, provide clear definitions of concepts, and indicate brain mechanisms for subsequent investigations.

Fresh alfalfa is difficult to ensile successfully because of its high moisture content and greater susceptibility to spoilage by Clostridia, Bacilli or Enterobacter. In this study, we evaluated the effects of mixing high-moisture alfalfa with peanut vine in different proportions on the bacterial communities and fermentation characteristics of silage. The high-moisture alfalfa and peanut vine were mixed at ratios of 10:0 (CK), 8:2 (TI), 7:3 (T2), 6:4 (T3) and 5:5 (T4), respectively. For each treatment, silos (25 × 35 cm) were anaerobically fermented in darkness at room temperature and analyzed after 45 days. The results showed that the CK silage was weakly fermented, as indicated by a low lactic acid concentration, a high pH value, and high levels of propionic acid (PA), butyric acid (BA) and ammonia nitrogen (NH3-N). As the proportion of peanut vine in the mixture increased, the pH level decreased, and levels of BA, propionic acid, NH3-N, crude protein(CP), nonprotein nitrogen and soluble protein also declined (p < 0.05), while the lactic acid concentration increased and levels of neutral detergent fiber (NDF) and water-soluble carbohydrates (WSC) also rose (p < 0.05). A protein component analysis of silage mixtures using the Cornell Net Carbohydrate and Protein System (CNCPS) showed that the content of the nonprotein nitrogen component (PA) decreased when the proportion of peanut vine increased, whereas the content of rapidly degraded protein (PB1) increased. Mixing with peanut vine also influenced the distribution of the bacterial community. Compared with the CK silage, the relative abundances of Enterococcus, Garciella and Anaerosporobacter in T2, T3 and T4 were significantly lower, while the relative abundance of Lactobacillus was significantly higher. In the T2, T3 and T4 groups, Garciella and Anaerosporobacter were not detected. In summary, in this study, we ensiled high-moisture alfalfa, which was weakly fermented. We found that mixing with peanut vine improved fermentation quality and optimized the structure of the bacterial community. Therefore, to improve the fermentation quality and nutritional value of silage, high-moisture alfalfa should be ensiled with at least 30% peanut vine.

To explore the molecular mechanisms of the antifungal compound phenazine-1-carboxamide (PCN) inhibits Rhizoctonia solani and discover potential targets of action, we performed an integrated analysis of transcriptome and metabolome in R. solani mycelium by whether PCN treating or not. A total of 511 differentially expressed genes (DEGs) were identified between the PCN treatment and control groups. The fluorescence-based quantitative PCR (qPCR) got the accordant results of the gene expression trends for ten randomly selected DEGs. The Gene Ontology (GO) enrichment analysis revealed that fatty acid metabolic process, fatty acid oxidation, and lipid oxidation were among the most enriched in the biological process category, while integral component of membrane, plasma membrane, and extracellular region were among the most enriched in the cellular component category and oxidoreductase activity, cofactor binding, and coenzyme binding were among the most enriched in the molecular function category. KEGG enrichment analysis revealed the most prominently enriched metabolic pathways included ATP-binding cassette (ABC) transporters, nitrogen metabolism, aminobenzoate degradation. The DEGs related functions of cellular structures, cell membrane functions, cellular nutrition, vacuole-mitochondrion membrane contact site and ATPase activity, pH, anti-oxidation, were downregulated. A total of 466 differential metabolites were found between the PCN treatment and control groups after PCN treatment. KEGG enrichment found purine, arachidonic acid, and phenylpropanoid biosynthesis pathways were mainly affected. Further results proved PCN decreased the mycelial biomass and protein content of R. solani, and superoxide dismutase (SOD) activity reduced while peroxidase (POD) and cytochrome P450 activities increased. The molecule docking indicted that NADPH nitrite reductase, ATP-binding cassette transporter, alpha/beta hydrolase family domain-containing protein, and NADPH–cytochrome P450 reductase maybe the particular target of PCN. In conclusion, the mechanisms via which PCN inhibits R. solani AG1IA may be related to cell wall damage, cell membrane impairment, intracellular nutrient imbalance, disturbed antioxidant system, and altered intracellular pH, which laid foundation for the further new compound designing to improve antifungal efficacy.

The objective of the present study was to investigate the effects of N-carbamylglutamate (NCG) supplementation on metabolic profile and microbiota in ruminal content and feces of lactating dairy cows under heat stress (HS). Forty-eight lactating Holstein cows (154 ± 13.6 days in milk) were assigned randomly to four treatments (n = 12), to receive 0, 15, 20, or 25 g/day of commercial NCG (proportion: 97.7%) for the period of 60 days. The recorded ambient temperature–humidity index (THI) suggested that the cows were exposed to HS for almost the entire experimental period (average THI: 80.6). Samples of ruminal content and feces were collected at the end of the trial (day 60) to determine the biological effects of NCG supplementation on metabolome and microbiota using mass spectrometry-based metabolomics and 16S rRNA gene sequencing techniques, respectively. Results showed that NCG supplementation enhanced the levels of ruminal microbial protein, total volatile fatty acids (VFAs), and the molar proportion of propionate in the rumen, but lowered the ruminal pH, ammonia nitrogen (NH3-N), and the ratio of acetate to propionate. NCG at doses of 20 and 25 g/day reduced the community richness and diversity of ruminal microbiota with the decrease of Shannon and Simpson diversity. Compositions of ruminal and fecal microbiotas were altered by NCG, and the PICRUSt results revealed that metabolic pathways of the bacteria, such as amino acid metabolism, energy metabolism, and pyruvate metabolism, were enriched in NCG groups. Distinct changes in the metabolomic profile of ruminal fluid were observed between the control and NCG groups. Changes of 26 metabolites mainly involved in arginine metabolism, glutamate metabolism, and nitrogen metabolism were observed associated with NCG supplementation. These results provided new insights into the effects of NCG on metabolomic profile and microbiota in ruminal content and feces, and the optimal dose of NCG supplemented to dairy cows was 20 g/hd/day, which contributed to understanding the effects of NCG on HS in lactating dairy cows.

The aim of this study was to investigate the effect of low-protein diets supplemented with rumen-protected lysine (RPLys) and methionine (RPMet) on growth performance, rumen fermentation, blood biochemical parameters, nitrogen metabolism, and gene expression related to N metabolism in the liver of Holstein bulls. Thirty-six healthy and disease-free Holstein bulls with a similar body weight (BW) (424 ± 15 kg, 13 months old) were selected. According to their BW, they were randomly divided into three groups with 12 bulls in each group in a completely randomized design. The control group (D1) was fed with a high-protein basal diet (CP13%), while bulls in two low-protein groups were supplied a diet with 11% crude protein and RPLys 34 g/d·head + RPMet 2 g/d·head (low protein with low RPAA, T2) or RPLys 55 g/d·head + RPMet 9 g/d·head (low protein with high RPAA, T3). At the end of the experiment, the feces and urine of dairy bulls were collected for three consecutive days. Blood and rumen fluid were collected before morning feeding, and liver samples were collected after slaughtering. The results showed that the average daily gain (ADG) of bulls in the T3 group was higher than those in D1 (p < 0.05). Compared with D1, a significantly higher nitrogen utilization rate (p < 0.05) and serum IGF-1 content (p < 0.05) were observed in both T2 and T3 groups; however, blood urea nitrogen (BUN) content was significantly lower in the T2 and T3 groups (p < 0.05). The content of acetic acid in the rumen of the T3 group was significantly higher than that of the D1 group. No significant differences were observed among the different groups (p > 0.05) in relation to the alpha diversity. Compared with D1, the relative abundance of Christensenellaceae_R-7_group in T3 was higher (p < 0.05), while that of Prevotellaceae _YAB2003_group and Succinivibrio were lower (p < 0.05). Compared with D1 and T2 group, the T3 group showed an expression of messenger ribonucleic acid (mRNA) that is associated with (CPS-1, ASS1, OTC, ARG) and (N-AGS, S6K1, eIF4B, mTORC1) in liver; moreover, the T3 group was significantly enhanced (p < 0.05). Overall, our results indicated that low dietary protein (11%) levels added with RPAA (RPLys 55 g/d +RPMet 9 g/d) can benefit the growth performance of Holstein bulls by reducing nitrogen excretion and enhancing nitrogen efficiency in the liver.

Abstract Background RNA-sequencing was performed to explore the bovine liver transcriptomes of Holstein cows to detect potential functional genes related to lactation and milk composition traits in dairy cattle. The bovine transcriptomes of the nine liver samples from three Holstein cows during dry period (50-d prepartum), early lactation (10-d postpartum), and peak of lactation (60-d postpartum) were sequenced using the Illumina HiSeq 2500 platform. Results A total of 204, 147 and 81 differentially expressed genes (DEGs, p

The objective of this study was to investigate the effects of dietary rumen-degradable starch (RDS, g/kg of DM) to rumen-degradable protein (RDP, g/kg of DM) ratios (SPR) on in vitro rumen fermentation characteristics and microbial protein synthesis (MCPS). Treatments were eight diets with SPR of 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5 and 2.6 and were formulated to be isoenergetic, isonitrogenous, and isostarch. Substrates were anaerobically incubated in sealed culture vials (100 mL) for 6, 24 or 48 h. Three incubation runs were conducted within two consecutive weeks. With the increase of the dietary SPR, the gas production (GP), in vitro dry matter disappearance (IVDMD) and concentration of MCPS and total volatile fatty acids (TVFA) linearly increased after 6 h of incubation (p ≤ 0.01), whereas they quadratically increased and peaked at the SPR of 2.3 after 24 and 48 h of incubation (p < 0.05). In response to dietary SPR increasing, the in vitro neutral detergent fiber disappearance (IVNDFD) quadratically increased (p < 0.01), and the ammonia nitrogen (NH3-N) concentration linearly decreased (p < 0.01) after 6, 24 and 48 h of incubation. Based on the presented results, an SPR of 2.3 is recommended for formulating a diet due to its greatest IVDMD, IVNDFD, GP, TVFA and MCPS. However, as the results obtained are strictly dependent on the in vitro conditions, further in vivo studies are needed to verify our findings.

Concrete pile foundations are the main load-carrying and aseismic structures for many large-scale building structures. Their structural quality will have a great impact on the safety of building structures. However, the imaging results still cannot easily meet practical demands in current ultrasonic Computerized Tomography (CT) imaging tests of concrete structures. In view of the current difficulties, a combinatorial optimization tomographic imaging method is proposed. First, a quadric broadening objective function with a clear physical meaning is established according to the characteristics of ultrasonic propagation in concrete. Then, a new CT imaging method of concrete pile foundation is formed by combining fast adaptive optimization search ability of Genetic Algorithm (GA) with the global search control ability of Simulated Annealing Algorithm (SAA). The numerical simulation experiments have shown that the usage of the correct priori information and the excellent characteristic of the Simulated Annealing Genetic Algorithm (SAGA) in searching for the global minimum value of the function have produced accurate and effective results with stable numerical values. Finally, the imaging method is verified by experiment, where results show that SAGA requires fewer iterations, has faster computation speed and gives more accurate imaging results compared with the single GA.

Rice grain yield is a complex and highly variable quantitative trait consisting of several key components, including the grain weight, the effective panicles per unit area, and the grain number per panicle (GNPP). The GNPP is a significant contributor to grain yield controlled by multiple genes (QTL) and is crucial for improvement. Attempts have been made to find genes for this trait, which has always been a challenging and arduous task through conventional methods. We combined a BSA analysis, RNA profiling, and a metabolome analysis in the present study to identify new candidate genes involved in the GNPP. The F2 population from crossing R4233 (high GNPP) and Ce679 (low GNPP) revealed a frequency distribution fitting two segregated genes. Three pools, including low, middle, and high GNPP, were constructed and a BSA analysis revealed six candidate regions spanning 5.38 Mb, containing 739 annotated genes. Further, a conjunctive analysis of BSA-Seq and RNA-Seq showed 31 differentially expressed genes (DEGs) in the candidate intervals. Subsequently, a metabolome analysis showed 1024 metabolites, with 71 significantly enriched, including 44 up and 27 downregulated in Ce679 vs. R4233. A KEGG enrichment analysis of these 31 DEGs and 71 differentially enriched metabolites (DEMs) showed two genes, Os12g0102100 and Os01g0580500, significantly enriched in the metabolic pathways’ biosynthesis of secondary metabolites, cysteine and methionine metabolism, and fatty acid biosynthesis. Os12g0102100, which encodes for the alcohol dehydrogenase superfamily and a zinc-containing protein, is a novel gene whose contribution to the GNPP is not yet elucidated. This gene coding for mitochondrial trans-2-enoyl-CoA reductase is involved in the biosynthesis of myristic acid, also known as tetradecanoic acid. The Os01g0580500 coding for the enzyme 1-aminoclopropane-1-carboxylate oxidase (OsACO7) is responsible for the final step of the ethylene biosynthesis pathway through the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) into ethylene. Unlike Os12g0102100, this gene was significantly upregulated in R4233, downregulated in Ce679, and significantly enriched in two of the three metabolite pathways. This result pointed out that these two genes are responsible for the difference in the GNPP in the two cultivars, which has never been identified. Further validation studies may disclose the physiological mechanisms through which they regulate the GNPP in rice.

In order to overcome the limitations of traditional ultrasonic imaging methods, which cannot directly give the imaging results of defects in a pipeline and have low imaging quality, a frequency domain synthetic aperture focusing pipeline imaging method based on phase migration is proposed. The measured data were reconstructed by frequency domain synthetic aperture focusing, and the results were compared with those of time-domain synthetic aperture focusing imaging. The results show that the frequency domain synthetic aperture imaging method is more accurate for the location and quantification of defects in a pipeline, the influence of noise waves on the imaging transverse resolution is reduced, and the imaging resolution is increased by nearly 30%. It was found that the quantitative errors of axial double defects are 5.2% and 17.9%, and that of circumferential defects is 7.5%. These results show that the frequency domain synthetic aperture focusing pipeline imaging algorithm based on phase migration can achieve the accurate determination of multiple defects in pipelines.

Glass fiber-reinforced plastics (GFRP) is widely used in many industrial fields. When acoustic emission (AE) technology is applied for dynamic monitoring, the interfering signals often affect the damage evaluation results, which significantly influences industrial production safety. In this work, an effective intelligent recognition method for AE signals from the GFRP damage is proposed. Firstly, the wavelet packet analysis method is used to study the characteristic difference in frequency domain between the interfering and AE signals, which can be characterized by feature vector. Then, the model of back-propagation neural network (BPNN) is constructed. The number of nodes in the input layer is determined according to the feature vector, and the feature vectors from different types of signals are input into the BPNN for training. Finally, the wavelet packet feature vectors of the signals collected from the experiment are input into the trained BPNN for intelligent recognition. The accuracy rate of the proposed method reaches to 97.5%, which implies that the proposed method can be used for dynamic and accurate monitoring of GFRP structures.

Gas sensors with excellent stability and a high response at room temperature has drawn a great deal of attention and demand for them is huge. Surface designs provide inspiration toward making more useful sensor devices. The facile electrospinning process and Ar plasma treatment are used to fabricate rich and stable oxygen vacancies that contain a core-shell structured SnO2 polyaniline (PANI) nanotube. It shows that the induced surface oxygen vacancies would accelerate the PANI shell to generate more protons, which can enhance its sensor responsibility through reacting with the target Ammonia (NH3) gas. It was also found that the obtained oxygen vacancies can be well-protected by the coated PANI shell, which enhance and stabilize the gas response. It shows that the room temperature for the gas response of NH3 can reach up to 35.3 at 100 ppm. Finally, its good stability is demonstrated by the response-recovery performances carried out over 3 months and multiple cycles. This work indicates that this well-designed PANI-coated plasma-treated SnO2 is a potential way to design ammonia gas sensors.

Previous research regarding Holstein cows has mainly focused on increasing milk yield. However, in order to maximize the economical profits of Holstein cattle farming, it is necessary to fully take advantage of Holstein bulls to produce high-grade beef. The present study aims to investigate different transcriptomic profiling of Holstein bulls and steers, via high-throughput RNA-sequencing (RNA-seq). The growth and beef quality traits of Holstein steers and bulls were characterized via assessment of weight, rib eye area, marbling score, shear force and intramuscular fat percentage of the longissimus lumborum (LL) muscle. The results indicated that castration improved the meat quality, yet reduced the meat yield. Subsequently, RNA-seq of the LL muscle from Holstein steers and bulls revealed a total of 56 differentially expressed genes (DEGs). We performed the functional enrichment analysis in Gene Ontology (GO) annotations of the DEGs using GOseq R package software and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis using KOBAS tool. Through the integrated analysis of DEGs with reported QTLs and SNPs, seven promising candidate genes potentially affecting the beef quality of LL muscle following castration were discovered, including muscle structural protein coding genes (MYH1, MYH4, MYH10) and functional protein coding genes (GADL1, CYP2R1, EEPD1, SHISA3). Among them, MYH10, GADL1, CYP2R1, EEPD1 and SHISA3 were novel candidate genes associated with beef quality traits. Notably, EEPD1 was associated with both meat quality and reproduction traits, thus indicating its overlapping role in responding to hormone change, and subsequently inducing beef quality improvement. Our findings provide a complete dataset of gene expression profile of LL in Holstein bulls and steers, and will aid in understanding how castration influence meat yield and quality.

Anthocyanin is a flavonoid compound with potential antioxidant properties beneficial to human health and sustains plant growth and development under different environmental stresses. In black rice, anthocyanin can be found in the stems, leaves, stigmas, and caryopsis. Although the anthocyanin biosynthesis in rice has been extensively studied, limited knowledge underlying the storage mechanism and transporters is available. This study undertook the complementation of computational and transcriptome analysis to decipher a potential multidrug and toxic compound extrusion (MATE) gene candidate for anthocyanin transportation in black rice caryopsis. The phylogenetic analysis showed that OsMATE34 has the same evolutionary history and high similarities with VvAM1, VvAM3, MtMATE2, SlMATE/MTP77, RsMATE8, AtFFT, and AtTT12 involved in anthocyanin transportation. RNA sequencing analysis in black caryopsis (Bc; Bc11, Bc18, Bc25) and white caryopsis (Wc; Wc11, Wc18, Wc25), respectively, at 11 days after flowering (DAF), 18 DAF, and 25 DAF revealed a total of 36,079 expressed genes, including 33,157 known genes and 2922 new genes. The differentially expressed genes (DEGs) showed 15,573 genes commonly expressed, with 1804 and 1412 genes uniquely expressed in Bc and Wc, respectively. Pairwise comparisons showed 821 uniquely expressed genes out of 15,272 DEGs for Wc11 vs. Bc11, 201 uniquely expressed genes out of 16,240 DEGs for Wc18 vs. Bc18, and 2263 uniquely expressed genes out of 16,240 DEGs for Wc25 vs. Bc25. Along with anthocyanin biosynthesis genes (OsPAL, OsCHS, OsCHI, OsF3H, OsDFR, OsANS, and OsUFGT/Os3GT), OsMATE34 expression was significantly upregulated in all Bc but not in Wc. OsMATE34 expression was similar to OsGSTU34, a transporter of anthocyanin in rice leaves. Taken together, our results highlighted OsMATE34 (Os08g0562800) as a candidate anthocyanin transporter in rice caryopsis. This study provides a new finding and a clue to enhance the accumulation of anthocyanin in rice caryopsis.

Anthocyanins belong to the group of flavonoid compounds broadly distributed in plant species responsible for attractive colors. In black rice (Oryza sativa L.), they are present in the stems, leaves, stigmas, and caryopsis. However, there is still no scientific evidence supporting the existence of compartmentalization and trafficking of anthocyanin inside the cells. In the current study, we took advantage of autofluorescence with anthocyanin’s unique excitation/emission properties to elucidate the subcellular localization of anthocyanin and report on the in planta characterization of anthocyanin prevacuolar vesicles (APV) and anthocyanic vacuolar inclusion (AVI) structure. Protoplasts were isolated from the stigma of black and brown rice and imaging using a confocal microscope. Our result showed the fluorescence displaying magenta color in purple stigma and no fluorescence in white stigma when excitation was provided by a helium–neon 552 nm and emission long pass 610–670 nm laser. The fluorescence was distributed throughout the cell, mainly in the central vacuole. Fluorescent images revealed two pools of anthocyanin inside the cells. The diffuse pools were largely found inside the vacuole lumen, while the body structures could be observed mostly inside the cytoplasm (APV) and slightly inside the vacuole (AVI) with different shapes, sizes, and color intensity. Based on their sizes, AVI could be grouped into small (Ф < 0.5 um), middle (Ф between 0.5 and 1 um), and large size (Ф > 1 um). Together, these results provided evidence about the sequestration and trafficking of anthocyanin from the cytoplasm to the central vacuole and the existence of different transport mechanisms of anthocyanin. Our results suggest that stigma cells are an excellent system for in vivo studying of anthocyanin in rice and provide a good foundation for understanding anthocyanin metabolism in plants, sequestration, and trafficking in black rice.