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The objective of this work is to investigate the chemical and nutritional value of milk thistle meal (MTM) in order to improve it and to provide theoretical support for its application in dairy cattle production. MTM was assessed in comparison with seven conventional protein feed sources, namely, soybean meal (SBM), cottonseed meal (CS), canola meal (CN), palm kernel meal (PK), rice bran meal (RB), corn germ meal (CG), and sesame meal (SS). The chemical composition of these feedstuffs was assessed using wet chemical analysis, the Cornell Net Carbohydrate and Protein System was used to evaluate the carbohydrate and protein fractions, and the in situ nylon bag technique and the modified three-step in vitro method were used to assess the rumen degradation and intestinal digestibility. Additionally, Fourier transform infrared technology was used to determine the feedstuff protein spectral molecular structure and its amino acid profile was also assessed. The result showed that MTM acid detergent fiber, lignin, unavailable nitrogen, and non-degradable carbohydrate content were higher than those of the other feedstuffs. It had a 17% and 36% rumen effective degradation rate of neutral detergent fiber and dry matter, respectively, and had the lowest small intestinal rumen undegradable protein digestibility rate. It was low in leucine, histidine, arginine, and proline, but high in methionine. The total area of amide I and amide II in the protein secondary structure was similar to that of CN and CS, and the amide I and II ratio was not different from that of RB. To sum up, MTM has a poor carbohydrate composition and is high in fiber but, in comparison to most other protein feeds, has a higher crude protein rumen effective degradation rate, similar to that of SBM, and it is a good source of methionine, a limiting amino acid. Hence, its nutritional value can be further improved for application in dairy feeding through processes such as microbial or enzymatic fermentation.

The aim of this study is to identify an alternative approach for simulating the in vitro fermentation and quantifying the production of rumen methane and rumen acetic acid during the rumen fermentation process with different total mixed rations. In this experiment, dietary nutrient compositions (neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), and dry matter (DM)) were selected as input parameters to establish three prediction models for rumen fermentation parameters (methane and acetic acid): an artificial neural network model, a genetic algorithm-bp model, and a support vector machine model. The research findings show that the three models had similar simulation results that aligned with the measured data trends (R 2 ≥ 0.83). Additionally, the root mean square errors (RMSEs) were ≤1.85 mL/g in the rumen methane model and ≤2.248 mmol/L in the rumen acetic acid model. Finally, this study also demonstrates the models' capacity for generalization through an independent verification experiment, as they effectively predicted outcomes even when significant trial factors were manipulated. These results suggest that machine learning-based in vitro rumen models can serve as a valuable tool for quantifying rumen fermentation parameters, guiding the optimization of dietary structures for dairy cows, rapidly screening methane-reducing feed options, and enhancing feeding efficiency.

This study aimed to determine changes and interactions of ruminal microbiota and chemical parameters in dairy cows fed FTMR. Twelve multiparous Holstein dairy cows (Body weight = 616 ± 13.4 kg; day in milk = 106 ± 7.55 d; and parity = 2.31 ± 0.49; mean ± standard deviation) were divided randomly into two treatments depending on the day in milk, milk production, and parity. The two treatments were: (1) total mixed ration (TMR) and (2) FTMR. Illumina MiSeq sequencing was used to explore the changes in the ruminal microbiota. The results revealed that the bacterial and fungal diversity of the FTMR group were significantly higher than the TMR group. The predominant microbiota phyla in the bacteria and fungi showed significant differences between TMR and FTMR, as follows: Verrucomicrobia ( p = 0.03) and Tenericutes ( p = 0.01), Ascomycota ( p = 0.04) and Basidiomycota ( p = 0.04). The dominant bacterial genera in the bacteria, fungi, protozoan, and archaea that showed significant differences between TMR and FTMR were Unclassified&#95; Bacteroidales ( p = 0.02), Unclassified&#95;RFP12 ( p = 0.03), Candida ( p = 0.0005), Bullera ( p = 0.002), Cryptococcus ( p = 0.007), and Ostracodinium ( p = 0.01). LefSe analysis was performed to reveal the biomarker genera of the rumen microbiota community (bacteria, fungi, protozoan, and archaea) in the TMR and FTMR were the genera Shuttleworthia , Ruminococcus, Cryptococcus , Mycosphaerella , Bullera , Candida , and Ostracodinium . NH 3 -N concentration ( p < 0.0001), total VFA concentration ( p = 0.003), and molar proportion in total VFA of acetate ( p = 0.01) were higher for the cows fed FTMR compared with the cows fed the TMR. Several bacterial genera showed significant correlations with rumen fermentation parameters. The genus Unclassified&#95; Bacteroidales and Bullera were positively correlated with total volatile fatty acids (VFA) and acetate, whereas Candida and Ostracodinium showed negative correlations. Meanwhile, propionate was positively correlated with Candida and negatively correlated with Bullera . The PICRUSt functional profile prediction indicated that the xenobiotics biodegradation and metabolism, the lipid, amino acid, terpenoids, and polyketides metabolisms of the FTMR group were significantly higher than that of the TMR group. The results imply that FTMR can increase lipid and amino acid metabolism, and modulate the rumen microbiome and improve ruminal fermentation.

Volatile fatty acids (VFAs) and methane are the main products of rumen fermentation. Quantitative studies of rumen fermentation parameters can be performed using in vitro techniques and machine learning methods. The currently proposed models suffer from poor generalization ability due to the small number of samples. In this study, a prediction model for rumen fermentation parameters (methane, acetic acid (AA), and propionic acid (PA)) of dairy cows is established using the stacking ensemble learning method and in vitro techniques. Four factors related to the nutrient level of total mixed rations (TMRs) are selected as inputs to the model: neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), and dry matter (DM). The comparison of the prediction results of the stacking model and base learners shows that the stacking ensemble learning method has better prediction results for rumen methane (coefficient of determination (R2) = 0.928, root mean square error (RMSE) = 0.968 mL/g), AA (R2 = 0.888, RMSE = 1.975 mmol/L) and PA (R2 = 0.924, RMSE = 0.74 mmol/L). And the stacking model simulates the variation of methane and VFAs in relation to the dietary fiber content. To demonstrate the robustness of the model in the case of small samples, an independent validation experiment was conducted. The stacking model successfully simulated the transition of rumen fermentation type and the change of methane content under different concentrate-to-forage (C:F) ratios of TMR. These results suggest that the rumen fermentation parameter prediction model can be used as a decision-making basis for the optimization of dairy cow diet compositions, rapid screening of methane emission reduction, feed beneficial to dairy cow health, and improvement of feed utilization.

The objective of this experiment was to explore the nutritional components of industrial hemp byproducts (industrial hemp ethanol extraction byproduct, IHEEB; industrial hemp stalk, IHS; industrial hemp seed meal, IHSM; industrial hemp oil filter residue, IHOFR) and provide theoretical support for the application of industrial hemp byproducts in dairy cattle production. This experiment used a combination of a wet chemical method with Cornell Net Carbohydrate and Protein System, in situ nylon bag technique, and three-step in vitro method to compare the chemical composition, carbohydrate and protein composition, in situ ruminal degradability and intestinal digestibility of industrial hemp byproducts and conventional feeds (alfalfa hay, AH; soybean meal, SBM). Available energy values were estimated based on the National Academies of Sciences, Engineering, and Medicine. The results showed that the nutritional composition of different feeds varied greatly. The two types of IHEEB were enriched with ash, crude protein (CP), neutral detergent fiber (NDF), and calcium, while the contents of neutral detergent insoluble crude protein, acid detergent insoluble crude protein, and acid detergent lignin were higher. As a result, the non-degradable carbohydrate and protein components were higher, and the effective degradation rate of rumen dry matter and protein was lower. IHS contains higher non-protein nitrogen and NDF, which enables it to provide more CP rumen effective degradation rate and carbohydrates, but the high acid detergent fiber also limits its application. IHSM possesses 296 g/kg CP and high rumen undegradable protein and intestinal digested protein, which can provide rumen bypass protein in dairy cows, making it a potentially good protein source. IHOFR had higher ether extract, rumen available protein degradation rate, and total tract digested protein, which can provide more energy and easily degradable protein for lactating cows. The available energy value of IHEEB and IHS was lower than AH, while SBM is between IHFOR and IHSM. In addition, the tetrahydrocannabinol of three industrial hemp byproducts that have not been assessed by the European Food Safety Authority (EFSA) was tested to evaluate their safety, and all of them were less than the limit set by ESFA. In conclusion, industrial hemp byproducts can be considered for inclusion in dietary formulations as unconventional feed sources for dairy cattle, but the purpose of use needs to be properly considered.

The purpose of this experiment was to explore the relationship of protein functional groups (including amide I, amide II, α-helix, and β-sheet) in byproducts of seed-used pumpkin (pumpkin seed cake, pumpkin seed coat, and seed-used pumpkin flesh) with their nutrient profiles and biodegradation characteristics. The experiment was designed to use conventional chemical analysis, combining the Cornell Net Carbohydrate and Protein System (CNCPS) and nylon bag technology to assess the nutritional value and biodegradation characteristics of seed-used pumpkin byproducts. Fourier transform infrared spectroscopy (FTIR) was used to analyze the protein molecular structure properties of byproducts of seed-used pumpkin. In this study, we also applied correlation and regression analysis. The results showed that different byproducts of seed-used pumpkin had different in situ biodegradation, nutrient supplies, and spectral structures in the protein region. Among the byproducts of seed-used pumpkin, acid detergent-insoluble crude protein (ADICP) and neutral detergent-insoluble crude protein (NDICP) contents of the pumpkin seed coat were the highest, resulting in the lowest effective degradabilities (EDs) of dry matter and crude protein. The crude protein (CP) ED values were ranked as follows: pumpkin seed cake > seed-used pumpkin flesh > pumpkin seed coat. Significant differences were observed in the peak areas of amide I and amide II and the corresponding peak heights in the two peak areas in the molecular structure of the protein. The peak areas of amide I and amide II and the corresponding peak heights were at the highest levels for pumpkin seed cake, whereas there was no significant difference between the pumpkin seed coat and seed-used pumpkin flesh. Similarly, the peak heights of α-helices and β-sheets were highest for pumpkin seed cake. Correlation and regression results indicated that amide I and amide II area and height, α-helix and β-sheet heights, and area ratios of amide I: amide II, as well as the height ratios of amide I: amide II, and α-helices: β-sheets effectively estimated nutrient supply and that the height ratio of α-helices: β-sheets was mostly sensitive to biodegradation characteristics in different byproducts of seed-used pumpkin. There were significant differences in CP chemical composition and digestibility of different byproducts of seed-used pumpkin that were strongly related to the changes in protein molecular structures.

The objective of this study was to investigate the fermentation quality and microbial community of corn stover (CS) or rice straw (RS) silage mixed with soybean curd residue (SCR). In this study, SCR and CS or RS were mixed at ratios of 75:25, 70:30, and 65:35, respectively, and measured for nutrient content, fermentation indices, and bacterial diversity after 30 days of ensiling. The results showed an increase in lactic acid (LA) concentration (p < 0.01) and crude protein (CP) content (p < 0.0001), a decrease in pH value (p < 0.01), the content of NDF (p < 0.01) and ADF (p < 0.01), and ammonia nitrogen (AN) concentration (p < 0.01) as the proportion of SCR in raw materials (CS or RS) increased. The addition of SCR to silage led to a decrease in bacterial diversity and contributed to an increased relative abundance of beneficial microorganisms, such as Lactobacillus, and a corresponding decrease in the relative abundance of undesirable microorganisms, such as Clostridium and Enterobacter. Collectively, the mixed silage of soybean curd residue with corn stover or rice straw preserved more nutrients and helped improve fermentation quality.

The objectives of this experiment were to investigate the effects of N -carbamylglutamate (NCG) on growth and slaughter performance, meat quality, nitrogen utilization, plasma antioxidant and amino acids of Holstein bulls. In this case, 24 Holstein bulls (490 ± 29.0 kg of body weights and 540 ± 6.1 d of age) were blocked by body weights and age and randomly assigned to 1 of 4 groups: (1) CON group: bulls were fed the control diet, (2) CON + NCG group: bulls were fed the control diet with 40 mg/kg BW NCG, (3) Urea group: bulls were fed the urea diet, and (4) Urea + NCG group: bulls were fed the urea diet with 40 mg/kg BW NCG. Feeding NCG significantly improved ADG, FCR, DM and CP digestibility, carcass weight, slaughter weight, DOP, eye muscle area, shear force ( p = 0.001) and reduced L* of color, drip loss and cooking loss. Concurrently, feeding the urea diet induced a decreased ADG, carcass weight and slaughter weight, DOP, eye muscle area and shear force. NCG decreased contents of fecal N and urinary N, plasma urea in bulls and ammonia but increased N retention and utilization, plasma NO, plasma Arg, Leu, Ile and Tyr. On the other hand, feeding the urea diet increased urinary N, plasma urea and ammonia. Thus the study efficiently demonstrates that beef benefited from being fed a NCG product in the urea diet by enhancing its growth and slaughter performance, meat quality, nitrogen metabolism and plasma amino acids.

The management of body condition score (BCS) during the dry period is associated with the postpartum health outcomes of dairy cows. However, the difference between the actual BCS and the fixed ideal value is not able to accurately predict the occurrence of postpartum diseases. This study aimed to use statistical process control (SPC) technology to monitor the BCS of dry cows, to evaluate the effect of control charts on nutritional strategies, and to explore the utility of SPC in predicting the incidence of postpartum subclinical ketosis (SCK). The BCS and SCK data of 286 cows from the dry off period to 60 days postpartum were collected to set up the early warning function. Three control charts, including a control chart for the average BCS of the herds, for the BCS of each dry cow, and for individual BCS, were established. The early warning signs for postpartum SCK development were: (1) an individual BCS more than 3.5 that remained unchanged for six weeks; (2) a capability index (CPK), an SPC tool, greater than -0.52. Using these parameters, the early warning signs of SCK development were verified in 429 dry cows. The results showed that the accuracy of early warning signal was 0.64 and the precision was 0.26. The control chart showed that the average BCS of dry cows was consistently higher than the expected upper limit of BCS during the experimental period, and that the addition of new cows to the herds increased the average BCS. In summary, the application of SPC technology to monitor the BCS of dry cows was not a good tool for the prediction of postpartum SCK occurrence but was an appropriate tool for guiding positive nutrition strategies.

Little information is available regarding the effect of supplementing 25-hydroxyvitamin D 3 during the transition period combined with a postpartum oral calcium bolus on Ca homeostasis. The objectives of the current study were to evaluate the effects of 25-hydroxyvitamin D 3 combined with postpartum oral calcium bolus on lactation performance, serum minerals and vitamin D 3 metabolites, blood biochemistry, and antioxidant and immune function in multiparous dairy cows. To evaluate the effects of 25-hydroxyvitamin D 3 combined with oral calcium, 48 multiparous Holstein cows were randomly assigned to one of four treatments: (1) supplementing 240 mg/day vitamin D 3 without a postpartum oral Ca bolus (control), (2) supplementing 240 mg/day vitamin D 3 with an oral Ca bolus containing 90 g of Ca immediately post-calving (Ca + VitD), (3) supplementing 6 g/day 25-hydroxyvitamin D 3 without an oral Ca bolus (25D), and (4) supplementing 6 g/day 25-hydroxyvitamin D 3 with an oral Ca bolus containing 90 g of Ca immediately post-calving (Ca + 25D). Lactation performance during the first 21 days was measured. Blood was collected at the initiation of calving and then 1, 2, 7, 14, and 21 days relative to the calving date. The yield of milk (0.05 < p < 0.10), energy-corrected milk ( p < 0.05), 3.5% fat-corrected milk ( p < 0.05), and milk protein ( p < 0.05) were significantly higher in 25-hydroxyvitamin D 3 -treated groups within 3 weeks of lactation than in vitamin D 3 -treated cows. The iCa ( p < 0.05) and tCa ( p < 0.05) were higher in both Ca and 25D + Ca cows than in the control and 25D groups within 48 h. The concentrations of serum tCa ( p < 0.05), tP ( p < 0.05), and 25-hydroxyvitamin D 3 ( p < 0.05) in 25D and 25D + Ca cows were higher than those in control and Ca cows within 21 days postpartum. Feeding 25-hydroxyvitamin D 3 also showed a lower concentration of malondialdehyde ( p < 0.05), interleukin 6 ( p < 0.05), and tumor necrosis factor-alpha (TNF-α) ( p < 0.05), as well as a higher concentration of alkaline phosphatase ( p < 0.05), total antioxidant capacity ( p < 0.05), and immunoglobulin G ( p < 0.05) than vitamin D 3 . Supplementing Ca bolus also showed lower concentrations of alanine transaminase ( p < 0.05) and TNF-α ( p < 0.05). In conclusion, supplementing 25-hydroxyvitamin D 3 during the transition period combined with a postpartum oral calcium bolus improved lactation performance, Ca homeostasis, and antioxidant and immune function of medium-production dairy cows within 21 days postpartum.

The objective of this experiment was to investigate the effect of lactic acid bacteria (LAB) and cellulase (CE) on the fermentation quality, rumen degradation rate and bacterial community of mixed silage of soybean residue (SR) and corn stover (CS). The experiment adopted a single-factor experimental design. Four treatment groups were set up: the control group (CON), lactic acid bacteria treatment group (LAB), cellulase treatment group (CE) and lactic acid bacteria + cellulase treatment group (LAB + CE). Among them, the amount of added LAB was 1 × 10 6 CFU/g, and the amount of added CE was 100 U/g. After 56 days of mixed silage, samples were taken and analyzed, and the chemical composition, fermentation quality, rumen degradation rate and microbial diversity were determined. The results showed that the pH of each treatment group was significantly ( p < 0.05) lower than that of CON, while the lactic acid and ammoniacal nitrogen contents of each treatment group were significantly higher than that of CON, with the highest contents in the LAB + CE group. The contents of DNFom (Ash-free NDF), ADFom (Ash-free ADF) and DM in the LAB + CE group were significantly lower than those in the CON group, while the content of crude protein (CP) was significantly higher than that in the CON group. The in situ effective degradation rates of DM (ISDMD), DNF (ISNDFD) and CP (ISCPD) were all significantly ( p < 0.05) higher in each treatment group than in the control group. The results of principal component analysis showed that the bacterial composition of the LAB, CE and LAB + CE groups was significantly different from that of the CON group ( p < 0.05). Bacterial genus level analysis showed that the content of lactic acid bacteria was significantly higher in the LAB + CE group than in the other treatment groups ( p < 0.05), while the content of undesirable bacteria was significantly lower than in the other treatment groups. The results showed that the addition of Lactobacillus and/or cellulase in mixed silage of SR and CS could effectively improve the quality of mixed silage fermentation, rumen degradation rate and microbial diversity, with better results when Lactobacillus and cellulase were added together, which provides new ideas for better application of SR and CS in dairy production.

N -carbamylglutamate (NCG), a structural analog of N -acetylglutamate, improves nitrogen utilization in dairy cows. However, the effects of NCG on bulls are unknown. The purpose of the current research was to investigate the effects of adding different amounts of NCG on growth performance, nutrient digestibility, nitrogen metabolism and plasma metabolites of fattening Holstein bulls. Twenty-four Holstein bulls with similar body weights (BW, 408 ± 21.9 kg) and ages (450 ± 6.1 d; all mean ± SD) were selected for the feeding trial. After 2 weeks of adaptation, bulls were blocked by BW and age and subsequently randomly assigned to 1 of 4 groups: (1) CON group (control diet), (2) L group (supplementation with 20 mg/kg BW NCG), (3) M group (supplementation with 40 mg/kg BW NCG), or (4) H group (supplementation with 80 mg/kg BW NCG). The addition of NCG linearly and quadratically increased the average daily gain (CON vs. L vs. M vs. H = 1.03 vs. 1.19 vs. 1.40 vs. 1.26 kg/d) ( p < 0.05), feed conversion ratio (CON vs. L vs. M vs. H = 11.92 vs. 9.22 vs. 7.76 vs. 8.62) ( p < 0.05), crude protein digestibility (CON vs. L vs. M vs. H = 64.3 vs. 63.8 vs. 67.7 vs. 65.8%) (0.05 < p < 0.10), N retention ( p < 0.05) and N utilization ( p < 0.05) of bulls, whereas the contents of fecal N (0.05 < p < 0.10) and urinary N (0.05 < p < 0.10) in NCG-fed bulls linearly decreased compared with those in CON bulls. Bulls fed NCG showed a quadratic increased plasma nitric oxide ( p < 0.05) concentration. Furthermore, Arg ( p < 0.05), Ile ( p < 0.05), Val ( p < 0.05), Ala ( p < 0.05), Glu ( p < 0.05), Ser ( p < 0.05), total essential amino acid ( p < 0.05) and total nonessential amino acid ( p < 0.05) concentrations linearly and quadratically increased with increasing doses of NCG. In contrast, plasma urea ( p < 0.05) and ammonia ( p < 0.05) concentration linearly and quadratically decreased with increasing doses of NCG. Overall, the addition of NCG increased plasma Arg, Ile, Val, TEAA and TNEAA concentration, which in turn resulted in a higher N utilization and, therefore, higher average daily gain in NCG-fed bulls, providing baseline data for the widespread application of NCG in beef cattle production.

The objective of this study was to investigate the effects of different additives on the fermentation quality, bacterial community, and aerobic stability of total mixed ration (TMR) silage containing wet corn gluten feed (WCGF) and corn stover. The TMR was ensiled with four treatments: (1) no additive (control); (2) lactic acid bacteria (LAB); (3) fibrolytic enzyme (EN); (4) LAB + EN. The EN and LAB + EN decreased the neutral detergent fiber and acid detergent fiber contents. Additives led to a higher lactic acid (LA) content ( p < 0.0001) compared to control at all ensiling times. Silages inoculated with LAB and LAB + EN had higher dry matter ( p = 0.0007), LA ( p < 0.0001) and acetic acid (AA) contents ( p < 0.0001) compared to control. The LAB and LAB + EN had significantly lowest ammonia nitrogen among the treatments, while no significant difference occurred after days 7 of ensiling. Silages treated with LAB and LAB + EN had a higher LAB count ( p < 0.0001) and a lower pH, yeast, and mold counts compared to other silages. The LAB and LAB + EN greatly increased the portions of Firmicutes and Lactobacillus ( p < 0.0001, and p < 0.0001, respectively) and reduced undesirable bacteria. Inoculation with LAB + EN and LAB improved aerobic stability of TMR silages indicated by higher and more stable LA and AA contents, smaller rise in pH, and yeast count than other silages. The LAB + EN and LAB reduced microbial diversity and improved the fermentation quality and aerobic stability of TMR silage containing WCGF and corn stover.

The objective of this research was to examine the effect of sodium formate (SF) and calcium propionate (CAP) on the fermentation characteristics and microbial community of wet brewers grains (WBG) after short-term storage. In the laboratory environment, fresh WBG was ensiled with (1) no additive (CON), (2) sodium formate (SF, 3 g/kg fresh weight), and (3) calcium propionate (CAP, 3 g/kg fresh weight) for 20 days. After opening, fermentation characteristics, chemical composition, rumen effective degradability, and the microbial community of ensiled WBG were analyzed. The addition of CAP had no effect on pH and lactic acid concentration and increased the concentrations of propionic acid; the SF group had the lowest pH and acetic acid, butyric acid, and ammonia nitrogen contents and the highest lactic acid concentration. After fermentation, the SF group had the highest contents of dry matter (DM), water-soluble carbohydrates (WSCs), and neutral detergent fiber (NDF). The contents of the three nutrients in the CAP group were significantly higher than those in the CON group. The addition of the two additives had little influence on the crude protein (CP) and acid detergent fiber (ADF) contents of the ensiled WBG. Two additives elevated in situ effective degradability of DM and NDF compared with the parameters detected in the CON group; WBG ensiled with SF had higher effective in situ CP degradability than that in the CON and CAP groups. The results of the principal component analysis indicate that the SF group and two other groups had notable differences in bacterial composition. The analysis of the genus level of the bacterial flora showed that the content of Lactobacillus in the SF group was significantly higher than that in the two other treatment groups, while the content of Clostridium was significantly lower than that in the two other treatment groups. Therefore, the addition of sodium formate can suppress the undesirable microorganisms, improve the fermentation qualities, and ensure that WBG is well preserved after 20 days of ensiling.

The objectives of this study were to investigate the ruminal bacterial changes during the feeding cycle. Six ruminally cannulated Holstein cows were used in this experiment. The high-forage (HF) and high-concentrate (HC) diets contained 70% and 30% dietary forage, respectively. Dairy cows were fed their respective diets for at least 28 days, then samples were collected at 0, 2, 4, 9, 12, 16 and 20 h post-feeding. The results showed that pH, the concentration of (total volatile fatty acids) TVFAs and the percentages of acetate, propionate and butyrate were significantly affected by diet and time interactions. The diversity of rumen microbiota in HF dietary treatments was significantly higher than that in the HC dietary treatments. ACE (Abundance-based Coverage Estimator) and Chao 1 indices peak at 12 h post-feeding and then decline over the next 8 h. The rumen microbiota was mainly composed of the phyla Firmicutes, Bacteroidetes and Proteobacteria without considering the diet and time. The Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) functional profile prediction indicated that the carbohydrate metabolism was different at 9, 12 and 20 h post-feeding time, which revealed that the soluble carbohydrates were enough for microbial fermentation shortly after feeding. This research gave a further explanation of the interactions among rumen microorganisms, which could further help manipulate the rumen metabolism., Competing Interests: All authors declare that they have no conflicts of interest.

: The objectives of this study were to investigate the difference in the mechanism of VFAs production combined with macrogenome technology under different forage-to-concentrate ratios and sampling times. Six ruminally cannulated Holstein cows were used in a randomized complete block design. The high forage (HF) and high concentrate (HC) diets contained 70 and 35% dietary forage, respectively. The results showed that pH was affected by sampling time, at 4 h after feeding had lower value. Excepted for acetate, the VFAs was increased with forage decreased. Propionate formation via the succinic pathway, in which succinate CoA synthetase (EC 6.2.1.5) and propionyl CoA carboxylase (EC 2.8.3.1) were key enzymes, and significantly higher in HC treatment than in HF treatment, Selenomonas , Ruminobacter , Prevotella , and Clostridium were the main microorganism that encodes these key enzymes. Butyrate formation via the succinic pathway, in which phosphate butyryltransferase (EC 2.3.1.19), butyrate kinase (EC 2.7.2.7) and pyruvate ferredoxin oxidoreductase (EC 1.2.7.1) are the important enzymes, Prevotella and Bacteroides played important role in encodes these key enzymes. This research gave a further explanation on the metabolic pathways of VFAs, and microorganisms involved in VFAs production under different F:C ration, which could further reveal integrative information of rumen function., Competing Interests: Supplementary Materials: The following are available online at www.mdpi.com/xxx/s1, Figure S1 The metagenomic abundance of key elements of the propionate production pathway. Table S1 Ingredient compositions and nutrition levels of diets (% of DM); Table S2 Primers used for real-time PCR quantification; The percentage of reads mapped to each enzyme involved in propionate propionate

This is a prospective observational study that evaluates the effects of body condition score (BCS) changes in primiparous Holstein cows during peripartum on their NEFA and BHBA concentrations, hormone levels, postpartum health, and production performance. The cows under study ( n = 213) were assessed to determine their BCS (5-point scale; 0.25-point increment) once a week during the whole peripartum by the same researchers; backfat was used for corrections. Blood samples were collected 21 and 7 days before calving and 7, 21, and 35 days after calving, and were assayed for NEFA, BHBA, growth hormone (GH), insulin, leptin, and adiponectin concentrations. The incidence of disease and milk yield were recorded until 84 days after calving. Cows were classified according to their BCS changes during peripartum as follows: Those that gained BCS (G; ΔBCS ≥ 0.25), maintained BCS (M; ΔBCS = 0-0.25), or lost BCS (L; ΔBCS ≥ 0.5). The BCS at -21 days and at 7, 14, and 21 days were different (p < 0.01), but trended toward uniformity in all groups at calving. The L group had higher NEFA and BHBA concentrations and hormone levels ( p < 0.01) than the M and G groups at 21 and 35 days after calving, and had a higher incidence of uterine and metabolic diseases; however, there were no differences in production performance between the various groups. In conclusion, a lower BCS in primiparous cows during peripartum influences the NEFA and BHBA concentrations, hormone levels, and occurrence of health problems postpartum. The postpartum effects of BCS changes appear prior to calving.

The purpose of this research was to evaluate whether relationships exist between odd- and branched-chain fatty acids (OBCFAs) originating from milk fat and the corresponding data of ruminal fermentation parameters, microbial populations, and base contents that were used to mark microbial protein in rumen. Nine lactating Holstein dairy cows with similar body weights and parity were selected in this study, and the samples of rumen and milk were collected at the early, middle, and late stages, respectively. The rumen and milk samples were collected over three consecutive days from each cow, and the ruminal and milk OBCFA profiles, ruminal fermentation parameters, bacterial populations, and base contents were measured. The results showed that the concentrations of OBCFAs, with the exception of C11:0 and C15:0, were significantly different between milk and rumen ( p < 0.05). The concentrations of anteiso -fatty acids in milk were higher than those in rumen, and the contents of linear odd-chain fatty acids were higher than those of branched-chain fatty acids in both milk and rumen. Significant relationships that existed between the concentrations of C11:0, iso- C15:0, anteiso- C15:0, C15:0, and anteiso- C17:0 in rumen and milk ( p < 0.05). The total OBCFA content in milk was positively related to the acetate molar proportion but negatively correlated with isoacid contents ( p < 0.05). The populations of Ruminococcus albus , R. flavefacients , and Eubacterium ruminantium were significantly related to milk C13:0 contents ( p < 0.05). The adenine/N ratio was negatively related to milk OBCFA content ( p < 0.05) but positively associated with the iso- C15:0/ iso- C17:0 ratio ( p < 0.05). Milk OBCFAs were significantly correlated with ruminal fermentation parameters, ruminal bacterial populations, and base contents. Milk OBCFAs had the potential to predict microbial nitrogen flow, and the prediction equations for ruminal microbial nitrogen flow were established for OBCFAs in dairy milk., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.