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Objective: To analyze the factors related to Helicobacter pylori (Hp) infection in patients with severe acute pancreatitis (SAP) and to observe the effect of Hp on SAP, and to provide a reference for future clinical prevention and treatment of Hp infection in SAP., Methods: A retrospective analysis was performed on 77 SAP patients admitted to Pingxiang People's Hospital between January 2020 and February 2022, with 33 Hp-infected individuals as the Hp-positive group and the other 44 patients being without Hp infection served as the Hp-negative group. First, the related factors of Hp infection in SAP patients were analyzed with multiple Logistic regression. Subsequently, the Acute Physiology and Chronic Health Evaluation II (APACHE II), Bedside Index for Severity in Acute Pancreatitis (BISAP) and Modified CT Severity Index (MCTSI) scores, as well as the levels of C-reactive protein (CRP), white blood cell (WBC), procalcitonin (PCT) and immunoglobulins A/M/G (IgA, IgM, and IgG) were recorded for inter-group comparisons. The adverse reactions and hospitalization time were also recorded. Besides, a six-month follow-up was carried out after discharge, and patients' quality of life was evaluated using the Short-Form 36 Item Health Survey (SF-36)., Results: Logistic regression analysis identified that history of Hp infection, long-term drinking, eating habits and history of biliary tract diseases were independent risk factors for Hp infection (all P<0.05). At 2 weeks after admission, higher APACHE II, BISAP and MCTSI scores were observed in Hp-positive group compared with Hp-negative group (all P<0.05). The Hp-positive group exhibited higher CRP, WBC and PCT levels while lower IgA, IgM and IgG levels during treatment compared to the Hp-negative group (all P<0.05). No difference was found in the incidence of adverse reactions between the two groups (P>0.05), but the hospitalization time of the Hp-positive group was significantly prolonged (P<0.05). The follow-up results determined better quality of life in the Hp-negative group, which resulted in higher SF-36 scores in various dimensions (P<0.05)., Conclusion: The history of Hp infection, long-term drinking, eating habits, and history of biliary tract diseases are all independent risk factors for Hp infection. Hp infection exacerbates disease progression of SAP, adversely influences patients' recovery, impairs their immune function, and compromises their prognoses., Competing Interests: None., (AJTR Copyright © 2024.)

Background: C16:0 and cis-9 C18:1 may have different effects on animal growth and health due to unique metabolism in vivo. This study was investigated to explore the different effects of altering the ratio of C16:0 and cis-9 C18:1 in fat supplements on growth performance, lipid metabolism, intestinal barrier, cecal microbiota, and inflammation in fattening bulls. Thirty finishing Angus bulls (626 ± 69 kg, 21 ± 0.5 months) were divided into 3 treatments according to the randomized block design: (1) control diet without additional fat (CON), (2) CON + 2.5% palmitic acid calcium salt (PA, 90% C16:0), and (3) CON + 2.5% mixed fatty acid calcium salt (MA, 60% C16:0 + 30% cis-9 C18:1). The experiment lasted for 104 d, after which all the bulls were slaughtered and sampled for analysis., Results: MA tended to reduce 0-52 d dry matter intake compared to PA (DMI, P = 0.052). Compared with CON and MA, PA significantly increased 0-52 d average daily gain (ADG, P = 0.027). PA tended to improve the 0-52 d feed conversion rate compared with CON (FCR, P = 0.088). Both PA and MA had no significant effect on 52-104 days of DMI, ADG and FCR (P > 0.05). PA tended to improve plasma triglycerides compared with MA (P = 0.077), significantly increased plasma cholesterol (P = 0.002) and tended to improve subcutaneous adipose weight (P = 0.066) when compared with CON and MA. Both PA and MA increased visceral adipose weight compared with CON (P = 0.021). Only PA increased the colonization of Rikenellaceae, Ruminococcus and Proteobacteria in the cecum, and MA increased Akkermansia abundance (P < 0.05). Compared with CON, both PA and MA down-regulated the mRNA expression of Claudin-1 in the jejunum (P < 0.001), increased plasma diamine oxidase (DAO, P < 0.001) and lipopolysaccharide (LPS, P = 0.045). Compared with CON and MA, PA down-regulated the ZO-1 in the jejunum (P < 0.001) and increased plasma LPS-binding protein (LBP, P < 0.001). Compared with CON, only PA down-regulated the Occludin in the jejunum (P = 0.013). Compared with CON, PA and MA significantly up-regulated the expression of TLR-4 and NF-κB in the visceral adipose (P < 0.001) and increased plasma IL-6 (P < 0.001). Compared with CON, only PA up-regulated the TNF-α in the visceral adipose (P = 0.01). Compared with CON and MA, PA up-regulated IL-6 in the visceral adipose (P < 0.001), increased plasma TNF-α (P < 0.001), and reduced the IgG content in plasma (P = 0.035). Compared with CON, PA and MA increased C16:0 in subcutaneous fat and longissimus dorsi muscle (P < 0.05), while more C16:0 was also deposited by extension and desaturation into C18:0 and cis-9 C18:1. However, neither PA nor MA affected the content of cis-9 C18:1 in longissimus dorsi muscle compared with CON (P > 0.05)., Conclusions: MA containing 30% cis-9 C18:1 reduced the risk of high C16:0 dietary fat induced subcutaneous fat obesity, adipose tissue and systemic low-grade inflammation by accelerating fatty acid oxidative utilization, improving colonization of Akkermansia, reducing intestinal barrier damage, and down-regulating NF-κB activation., (© 2024. The Author(s).)

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 gut microbiome plays an important role in the healthy and efficient farming of dairy cows. However, high-dimensional microbial information is difficult to interpret in a simplified manner. We collected fecal samples from 161 cows and performed 16S amplicon sequencing. We developed an interpretable machine learning framework to classify individuals based on their milk urea nitrogen (MUN) concentrations. In this framework, we address the challenge of handling high-dimensional microbial data imbalances and identify 9 microorganisms strongly correlated with MUN. We introduce the Shapley Additive Explanations (SHAP) method to provide insights into the machine learning predictions. The results of the study showed that the performance of the machine learning model improved (accuracy = 72.7%) after feature selection on high-dimensional data. Among the 9 microorganisms, g&#95;&#95;Firmicutes&#95;unclassified had the greatest impact in the model. This study provides a reference for precision animal husbandry., Competing Interests: The authors declare no competing interests., (© 2024 The Authors.)

This study evaluated the effects of different proportions of palmitic (C16:0) and oleic ( cis -9 C18:1) acids in fat supplements on rumen fermentation, glucose (GLU) and lipid metabolism, antioxidant function, and visceral fat fatty acid (FA) composition in Angus bulls. The design of the experiment was a randomized block design with 3 treatments of 10 animals each. A total of 30 finishing Angus bulls (21 ± 0.5 months) with an initial body weight of 626 ± 69 kg were blocked by weight into 10 blocks, with 3 bulls per block. The bulls in each block were randomly assigned to one of three experimental diets: (1) control diet without additional fat (CON), (2) CON + 2.5% palmitic calcium salt (PA; 90% C16:0), (3) CON + 2.5% mixed FA calcium salts (MA; 60% C16:0 + 30% cis -9 C18:1). Both fat supplements increased C18:0 and cis -9 C18:1 in visceral fat ( P  < 0.05) and up-regulated the expression of liver FA transport protein 5 ( FATP5 ; P  < 0.001). PA increased the insulin concentration ( P  < 0.001) and aspartate aminotransferase activity (AST; P  = 0.030) in bull's blood while reducing the GLU concentration ( P  = 0.009). PA increased the content of triglycerides (TG; P  = 0.014) in the liver, the content of the C16:0 in visceral fat ( P  = 0.004), and weight gain ( P  = 0.032), and up-regulated the expression of liver diacylglycerol acyltransferase 2 ( DGAT2 ;  P  < 0.001) and stearoyl-CoA desaturase 1 ( SCD1 ; P  < 0.05). MA increased plasma superoxide dismutase activity (SOD; P  = 0.011), reduced the concentration of acetate and total volatile FA (VFA) in rumen fluid ( P  < 0.05), and tended to increase plasma non-esterified FA (NEFA; P  = 0.069) concentrations. Generally, high C16:0 fat supplementation increased weight gain in Angus bulls and triggered the risk of fatty liver, insulin resistance, and reduced antioxidant function. These adverse effects were alleviated by partially replacing C16:0 with cis -9 C18:1., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2024 The Authors.)

Aerosol ammonium (NH 4 + ), mainly produced from the reactions of ammonia (NH 3 ) with acids in the atmosphere, has significant impacts on air pollution, radiative forcing, and human health. Understanding the source and formation mechanism of NH 4 + can provide scientific insights into air quality improvements. However, the sources of NH 3 in urban areas are not well understood, and few studies focus on NH 3 /NH 4 + at different heights within the atmospheric boundary layer, which hinders a comprehensive understanding of aerosol NH 4 + . In this study, we perform both field observation and modeling studies (the Community Multiscale Air Quality, CMAQ) to investigate regional NH 3 emission sources and vertically resolved NH 4 + formation mechanisms during the winter in Beijing. Both stable nitrogen isotope analyses and CMAQ model suggest that combustion-related NH 3 emissions, including fossil fuel sources, NH 3 slip, and biomass burning, are important sources of aerosol NH 4 + with more than 60% contribution occurring on heavily polluted days. In contrast, volatilization-related NH 3 sources (livestock breeding, N-fertilizer application, and human waste) are dominant on clean days. Combustion-related NH 3 is mostly local from Beijing, and biomass burning is likely an important NH 3 source (∼15%-20%) that was previously overlooked. More effective control strategies such as the two-product (e.g., reducing both SO 2 and NH 3 ) control policy should be considered to improve air quality., Competing Interests: Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2024 Science China Press. Published by Elsevier B.V. All rights reserved.)

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.

Little information exists on the variation in morphological characteristics, nutritional value, ruminal degradability, and molecular structural makeup of diverse whole-plant silage corn (WPSC) cultivars among different growing regions. This study investigated the between-regions (Beijing, Urumchi, Cangzhou, Liaoyuan, Tianjin) discrepancies in five widely used WPSC cultivars in China (FKBN, YQ889, YQ23, DK301 and ZD958), in terms of 1) morphological characteristics; 2) crude protein (CP) chemical profile; 3) Cornell Net Carbohydrate and Protein System (CNCPS) CP subfractions; 4) in situ CP degradation kinetics; and 5) CP molecular structures. Our results revealed significant growing region and WPSC cultivar interaction for all estimated morphological characteristics (P < 0.001), CP chemical profile (P < 0.001), CNCPS subfractions (P < 0.001) and CP molecular structural features (P < 0.05). Except ear weight (P = 0.18), all measured morphological characteristics varied among different growing regions (P < 0.001). Besides, WPSC cultivars planted in different areas had remarkably different CP chemical profiles and CNCPS subfractions (P < 0.001). All spectral parameters of protein primary structure of WPSC differed (P < 0.05) due to the growing regions, except amide II area (P = 0.28). Finally, the area ratio of amide I to II was negatively correlated with the contents of soluble CP (δ = -0.66; P = 0.002), CP (δ = -0.61; P = 0.006), non-protein nitrogen (δ = -0.56; P = 0.004) and acid detergent insoluble CP (δ = -0.43; P = 0.008), in conjunction with a positive correlation with moderately degradable CP (PB1; δ = 0.58; P = 0.01). In conclusion, the cultivar of DK301 exhibited high and stable CP content. The WPSC planted in Beijing showed high CP, SCP and NPN. The low rumen degradable protein of WPSC was observed in Urumchi. Meanwhile, above changes in protein profiles and digestibility were strongly connected with the ratio of amide I and amide II., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Animal nutritionists have incessantly worked towards providing livestock with high-quality plant protein feed resources. Soybean meal (SBM) has been an essential and predominantly adopted vegetable protein source in livestock feeding for a long time; however, several SBM antinutrients could potentially impair the animal's performance and growth, limiting its use. Several processing methods have been employed to remove SBM antinutrients, including fermentation with fungal or bacterial microorganisms. According to the literature, fermentation, a traditional food processing method, could improve SBM's nutritional and functional properties, making it more suitable and beneficial to livestock. The current interest in health-promoting functional feed, which can enhance the growth of animals, improve their immune system, and promote physiological benefits more than conventional feed, coupled with the ban on the use of antimicrobial growth promoters, has caused a renewed interest in the use of fermented SBM (FSBM) in livestock diets. This review details the mechanism of SBM fermentation and its impacts on animal health and discusses the recent trend in the application and emerging advantages to livestock while shedding light on the research gap that needs to be critically addressed in future studies. FSBM appears to be a multifunctional high-quality plant protein source for animals. Besides removing soybean antinutrients, beneficial bioactive peptides and digestive enzymes are produced during fermentation, providing probiotics, antioxidants, and immunomodulatory effects. Critical aspects regarding FSBM feeding to animals remain uncharted, such as the duration of fermentation, the influence of feeding on digestive tissue development, choice of microbial strain, and possible environmental impact., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2024 The Authors.)

This study used Silibinin as an additive to conduct fermentation experiments, wherein its effects on rumen gas production, fermentation, metabolites, and microbiome were analyzed in vitro . The silibinin inclusion level were 0 g/L (control group), 0.075 g/L, 0.15 g/L, 0.30 g/L, and 0.60 g/L (experimental group). Fermentation parameters, total gas production, carbon dioxide (CO 2 ), methane (CH 4 ), hydrogen (H 2 ), and their percentages were determined. Further analysis of the rumen microbiome's relative abundance and α/β diversity was performed on the Illumina NovaSeq sequencing platform. Qualitative and quantitative metabolomics analyses were performed to analyze the differential metabolites and metabolic pathways based on non-targeted metabolomics. The result indicated that with an increasing dose of silibinin, there was a linear reduction in total gas production, CO 2 , CH 4 , H 2 and their respective percentages, and the acetic acid to propionic acid ratio. Concurrent with a linear increase in pH, when silibinin was added at 0.15 g/L and above, the total volatile fatty acid concentration decreased, the acetic acid molar ratio decreased, the propionic acid molar ratio increased, and dry matter digestibility decreased. At the same time, the relative abundance of Prevotella , Isotricha , Ophryoscolex , unclassified&#95;Rotifera , Methanosphaera , Orpinomyces , and Neocallimastix in the rumen decreased after adding 0.60 g/L of silibinin. Simultaneously, the relative abundance of Succiniclasticum , NK4A214&#95;group , Candidatus&#95;Saccharimonas , and unclassified&#95;Lachnospiraceae increased, altering the rumen species composition, community, and structure. Furthermore, it upregulated the ruminal metabolites, such as 2-Phenylacetamide, Phlorizin, Dalspinin, N6-(1,2-Dicarboxyethyl)-AMP, 5,6,7,8-Tetrahydromethanopterin, Flavin mononucleotide adenine dinucleotide reduced form (FMNH), Pyridoxine 5'-phosphate, Silibinin, and Beta-D-Fructose 6-phosphate, affecting phenylalanine metabolism, flavonoid biosynthesis, and folate biosynthesis pathways. In summary, adding silibinin can alter the rumen fermentation parameters and mitigate enteric methane production by regulating rumen microbiota and metabolites, which is important for developing novel rumen methane inhibitors., Competing Interests: YS was employed by Beijing Sunlon Livestock Development Company Limited. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Liu, Shen, Ma, Li, Lambo, Dai, Shen, Qu and Zhang.)

This study evaluated the effects of changing the ratio of palmitic, stearic, and oleic acids in dietary fat on nutritional metabolism, growth performance, and meat quality of finishing Angus bulls. Bulls received the following three treatments: (1) a control diet without fat supplement (CON), (2) CON + mixed fatty acid supplement (58% C16:0 + 28% cis-9 C18:1; MIX), (3) CON + saturated fatty acid supplement (87% C16:0 + 10% C18:0; SFA). In summary, both fat treatment diets simultaneously increased saturated fatty acids C16:0 (P = 0.025), C18:0 (P < 0.001) and total monounsaturated fatty acids (P = 0.008) in muscle, thus balancing the ratio of unsaturated to saturated fatty acids in muscle. MIX diet increased the digestibility of dry matter (P = 0.014), crude protein (P = 0.038), and ether extract (P = 0.036). SFA diet increased the daily gain (P = 0.032) and intramuscular fat content (P = 0.043). The high content of C16:0 and C18:0 in the SFA diet promoted weight gain and fat deposition of beef cattle by increasing feed intake, up-regulating the expression of lipid uptake genes and increasing deposition of total fatty acids, resulting in better growth performance and meat quality., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest in the study., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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.

Background: Unreasonable use of antibiotics in animals is a major concern and will remain so, thus affecting people's health. However, the application of plant extracts can better solve this problem., Objectives: The purpose of this study was to study the effect of Moringa leaf flavonoids on the production performance, immunity, and rumen fermentation of dairy cows., Methods: Nine Holstein multiparous cows (average weight: 550 kg; days of lactation: 150 ± 6 days) were used in the experiment, using a 3 × 3 Latin square design. Cows were divided into three groups, each of which was supplemented with 0, 50, or 100 mg/body weight (BW) Moringa oleifera leaf flavonoids. Each experimental period consisted of three periods of 21 days, and the prefeeding period lasted 15 days., Results: Our results indicated that supplementation with Moringa leaf flavonoids increased the protein content and decreased the number of somatic cells in milk; had little effect on the biochemical indices of blood, the rumen fermentation, and serum biochemical indicators; and improved the activity of antioxidant enzymes, the antioxidant capacity, and immunity., Conclusions: Addition of 50 mg/BW Moringa leaf flavonoids to cow enhanced the antioxidant and immunity capacity in dairy cows but did not affect physiological levels of common biochemical parameters in blood or fermentation parameters in rumen., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

This research aimed to investigate the effects of replacing soybean meal with high-oil pumpkin seed cake (HOPSC) on ruminal fermentation, lactation performance, milk fatty acid, and ruminal bacterial community in Chinese dairy cows. Six multiparous Chinese Holstein cows at 105.50 ± 5.24 d in milk (mean ± standard deviation) and 36.63 ± 0.74 kg/d of milk yield were randomly allocated, in a 3 × 3 Latin square design, to 3 dietary treatments in which HOPSC replaced soybean meal. Group 1 was the basal diet with no HOPSC (0HOPSC); group 2 was a 50% replacement of soybean meal with HOPSC and dried distillers grains with solubles (DDGS; 50HOPSC), and group 3 was a 100% replacement of soybean meal with HOPSC and DDGS (100HOPSC). We found no difference in the quantity of milk produced or milk composition among the 3 treatment groups. Feed efficiency tended to increase linearly as more HOPSC was consumed. In addition, rumen fermentation was not influenced when soybean meal was replaced with HOPSC and DDGS; the relative abundance of ruminal bacteria at the phylum and genus levels was altered. We also observed that as the level of HOPSC supplementation increased, the relative abundance of Firmicutes and Tenericutes linearly increased, whereas that of Bacteroidetes decreased. However, with increasing HOPSC supplementation, the relative abundance of Ruminococcus decreased linearly at the genus level in the rumen, and the relative abundance of Prevotella showed a linear downward tendency. Changes in dietary composition and rumen bacteria had no significant effect on the fatty acid composition of milk. In conclusion, our results indicated that replacing soybean meal with a combination of HOPSC and DDGS can meet the nutritional needs of high-yielding dairy cows without adversely affecting milk yield and quality; however, the composition of rumen bacteria could be modified. Further study is required to investigate the effects of long-term feeding of HOPSC on rumen fermentation and performance of dairy cows., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

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.

This trial was designed to investigate the effects of industrial hemp ethanol extraction byproduct (IHEEB) and Chinese wildrye hay (CWH) replacement of alfalfa hay (AH) on digestibility, and lactation performance, plasma metabolites, ruminal fermentation, and bacterial communities in Holstein dairy cows. Nine healthy multiparous Holstein cows (parity = 3) with similar body weights (584 ± 12.3 kg), days in milk (108 ± 11.4), and milk yields (30 ± 1.93 kg; all mean ± standard deviation) were used in a replicated 3 × 3 Latin square design with 3 periods of 21 d. During each period, each group consumed 1 of 3 diets: (1) 0% IHEEB (0IHEEB); (2) 6.0% IHEEB and 1.7% Chinese wildrye hay (6IHEEB); (3) 10.8% IHEEB and 4.3% Chinese wildrye hay (11IHEEB). The diets in each group were isocaloric and isonitrogenous, with similar contents of concentrate and silage but different ratios of IHEEB and CWH to replace AH. The results showed that increasing the substitute did not affect the total-tract apparent nutrient digestibility. There was no difference in lactation performance of dairy cows fed the three diets, except for the cows' somatic cell count (SCC), which decreased with the increase in the amount of the substitute. Cannabidiol and tetrahydrocannabinol were not detected in milk samples of dairy cows in the different treatment groups. 6IHEEB and 11IHEEB-fed cows showed a linear decrease in total volatile fatty acids (VFA) and butyrate compared to the 0IHEEB cows. Plasma IL-1β content quadratically decreased with feeding IHEEB and CWH, and other blood parameters were unaffected. The rumen fluid's relative abundances of Bacteroidota, Fibrobacterota , and Prevotellaceae quadratically increased, while Firmicutes tended to decrease quadratically as the substitution increased. Feeding IHEEB and CWH linearly increased the relative abundances of Firmicutes, Lachnospiraceae, Monoglobaceae , and Butyricicoccaceae in the feces. As the substitution increased, the cost of dairy farming was reduced. In summary, substituting AH with IHEEB and CWH in diets did not affect the total-tract apparent nutrient digestibility, improved milk composition, and plasma immune indices. It changed the bacterial composition in rumen fluid and feces and improved dairy farming benefits., Competing Interests: JH was employed by Heilongjiang Wellhope Agri-Tech Co., Ltd. PH was employed by Harbin Wellhope Trading Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wang, Yu, Wang, Song, Lambo, Huang, He, Li and Zhang.)

The mitigation and prevention of acute immune stress are essential for livestock production. Clostridium butyricum ( C. butyricum ) has shown positive effects in stabilizing intestinal microbiota disorders, improving immune function and inhibiting disease development, but its effects on ruminants are unclear. Therefore, the current trial hypothesized that C. butyricum could improve goats' immune function and antioxidant capacity by regulating bacterial communities and blood metabolism and effectively alleviating the acute immune stress induced by Lipopolysaccharides (LPS). Sixteen healthy goats were fed C. butyricum for 70 days, and the goats were challenged with LPS on day 71. Blood and feces were collected at 0 h and 6 h after the challenge to evaluate the effects of C. butyricum on their intestinal microbiota, immune function, antioxidant function, and plasma metabolites. The results showed that C. butyricum had no significant effect on plasma biochemical parameters at the beginning of the LPS challenge. However, supplementation with C. butyricum increased plasma levels of IgA, IgG, T-SOD, and T-AOC ( P < 0.05), but TNF-α , IL-6 , and MDA were decreased ( P < 0.05). In contrast, IL-10 showed an increasing trend ( P < 0.10) . Rectal microbiota analysis showed that C. butyricum significantly increased the relative abundance of Epsilonbacteraeota at the phylum level of goats; at the genus level, the relative abundances of Campylobacter and Anaerorhabdus]&#95;furcosa&#95;group were also significantly increased ( P < 0.05 ). Christensenellaceae&#95;R-7&#95;group as the dominant microbiota also showed a significant increase in their abundance values, while Clostridium and Lachnospiraceae&#95;UCG-001 were significantly lower ( P < 0.05 ). When the LPS challenge continued up to 6 h, dietary supplementation with C. butyricum still resulted in significantly higher plasma concentrations of IgA, IL-10 , and T-SOD in goats than in the control group, reducing TNF-α levels ( P < 0.05 ). In addition, plasma levels of T-CHOL and LDL were significantly reduced, and the expression of d-proline was significantly upregulated according to metabolomic analysis ( P < 0.05 ). In conclusion, dietary supplementation with C. butyricum helped optimize the expression of bacterial communities and plasma metabolites to enhance the ability of goats to alleviate acute immune stress., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhang, Hou, Wang, Yu, Zhang and Sun.)

The objective of this study was to investigate the effect of waste milk with antibiotic residue on rumen fermentation and rumen bacterial composition of dairy calves during pre-weaned and post-weaned periods. A total of 24 Holstein male calves (43.4 ± 0.93 kg body weight, mean ± standard error) were allocated into four blocks based on birth date. Dairy calves were supplied 100% milk replacer (MR, n  = 8), 50% milk replacer mixed with 50% waste milk (MM, n = 8), or 100% waste milk (WM, n = 8). Ruminal samples were collected at 49 and 63 days of age and then subjected to determinations of pH value, volatile fatty acids (VFA), ammonia nitrogen (NH 3 -N) and 16S rRNA gene amplicon sequencing. The results showed that feeding WM had no effect on the pH value, the concentrations of VFA (acetic acid, propionic acid, butyric acid, isovaleric acid, valeric acid), and NH 3 -N in dairy calves compared to feeding MR. However, from 49 to 63 days of age, the pH value ( p  < 0.001) was significantly increased, while the levels of total VFA ( p  = 0.004), acetic acid ( p  = 0.01), propionic acid ( p  = 0.003) and valeric acid ( p  < 0.001) were significantly decreased. For rumen microorganisms, there was no differences in bacterial diversity among the treatments. But the relative abundance of Veillonellaceae was significantly lower ( p  = 0.05) in the calves fed WM than that from MR group at 49 days of age; however, no difference was detected at 63 days of age. Feeding WM to calves tended to reduce family Veillonellaceae and genus Olsenella in the rumen at 49 days of age ( p  = 0.049). Analysis of temporal changes in rumen bacteria based on alpha-diversity and beta-diversity as well as the microbial relative abundances did not exhibit any difference. In addition, relative abundances of Clostridia&#95;UCG-014 , Prevotella , Syntrophococcus , Eubacterium&#95;nodatum&#95;group , Pseudoramibacter and Solobacterium were correlated with rumen pH value and the concentrations of TVFA, propionic acid, isovaleric acid, valeric acid and NH 3 -N. In conclusion, compare to MR, calves supplied with WM had little changes on the rumen pH value, NH 3 -N or VFAs contents. Additionally, limited effects could be found on rumen microbiota in the calves fed WM. However, further studies needed to explore if there exist any long-term effects of early-life rumen microbiota modulation on dairy cows., Competing Interests: CL was employed by Harbin Wondersun the Cow Feeds the Reproduction Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhang, Cheng, Lv, Bai, Sun, Liu, Liu, Zhang and Xin.)

Oat hay and alfalfa hay are important roughage resources in livestock production. However, the effect of the mixture of oat hay and alfalfa hay on the meat quality of Albas goats is unclear. This study aimed to investigate the effects of feeding different proportions of oat hay and alfalfa hay on the growth performance and meat quality of Albas goats. Therefore, 32 goats were fed for 70 days and randomly divided into four treatment groups on the principle of similar weight: whole oat group (OAT), oat alfalfa ratio 3:7 group (OA73), oat alfalfa ratio 7:3 group (OA37) and whole alfalfa group (Alfalfa), with eight goats in each group. Daily feed intake records, feces, feed samples, and rumen fluid collection were made throughout the trial. The goats were weighed on the last day of the trial, and four goats per group were randomly selected for slaughter. Cecum contents, meat samples, and hot carcass weight were collected, and data were recorded. Furthermore, the relationship between the rumen and cecal microbes on performance and meat quality was clarified by analyzing the rumen and hindgut microbiomes. The results showed that feeding alfalfa could significantly reduce the daily weight gain of fattening goats. Compared with the highest group (OA37), the daily weight gain decreased by 19.21%. Although there was no significant change in feed intake in the four treatments, the feed conversion rate of the alfalfa group significantly decreased by 30.24-36.47% compared to the other groups. However, with the increased alfalfa content, MDA decreased significantly, T-AOC was up-regulated, and the antioxidant activity of the fattened goat meat fed with the high alfalfa group was significantly higher than that of the low alfalfa group. Notably, the abundance of Bacteroidales&#95;unclassified and Clostridium were strongly correlated with T-AOC and MDA. Therefore, increasing the proportion of alfalfa in the diet can affect the antioxidant activity of goat meat by improving the gut microbiota, while an oat-hay mixture can improve the growth performance of livestock., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Sun, Hou, Yu, Zhang, Zhang and Xu.)

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.

Clostridium butyricum , as a probiotic with a variety of active products, has been widely used to improve the intestinal health of humans and animals. Previous studies had demonstrated that Clostridium butyricum exhibited potential protective and positive effects in human disease research and animal production by producing a variety of beneficial substances, such as intestinal inflammation, the intestinal epithelial barrier, metabolic diseases, and regulation of the gut microbiota. Therefore, we hypothesized that dietary Clostridium butyricum supplementation could improve gut health in fattening goats by modulating gut microbiota. However, it is unclear whether Clostridium butyricum can reach the intestine through the rumen, so 15 healthy Albas goats were selected and randomly divided into 3 treatments with 5 replicates in each group. The groups were divided as follows: control group (CON: basal diet), rumen-protected Clostridium butyricum group (RPCB: basal diet plus 1.0 × 10 9 CFU/kg Clostridium butyricum coated with hydrogenated fat), and Clostridium butyricum group (CB: basal diet plus 1.0 × 10 9 CFU/kg Clostridium butyricum ). The experiment was slaughtered after a 70-day growth test, and the jejunal mucosa and intestinal contents of the goats were collected to determine tight junction proteins related genes expression and 16S rDNA microbial sequencing analysis to evaluate the intestine health. The results showed that dietary supplementation with Clostridium butyricum significantly increased the expression of the Claudin-4 gene of the jejunal mucosa ( P < 0.05) and had a trend toward a significant increase in the Occludin gene (0.05 < P < 0.10). However, Clostridium butyricum had no significant effect on the expression of intestinal inflammatory factors ( P > 0.10). In addition, the relative fractionation of Clostridium and Clostridiaceae&#95;unclassified in the gut microbiota at the genus level decreased significantly compared with controls ( P < 0.05). The results of the analysis of the level of Clostridium species showed that Clostridium butyricum only existed in the treatment group. And the correlation results showed that Occludin and Claudin-4 genes were positively correlated with Sharppea and Clostridium butyricum , and negatively correlated with Clostridium ( P < 0.05 ). Supplementing Clostridium butyricum in the diet did not significantly affect the intestinal immune function of goats, while regulation of the intestinal microbiota was associated with improving the intestinal epithelial barrier., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zhang, Hou, Yu, Wang, Ni, Zi, Xin, Zhang and Sun.)

This study was conducted to evaluate the effect of dietary supplementation with lysophospholipids (LPLs) on the growth performance, nutrient digestibility, nitrogen utilization, and blood metabolites of finishing beef cattle. In total, 40 Angus beef bulls were blocked for body weight (447 ± 9.64 kg) and age (420 ± 6.1 days) and randomly assigned to one of four treatments (10 beef cattle per treatment): (1) control (CON; basal diet); (2) LLPL (CON supplemented with 0.012% dietary LPL, dry matter (DM) basis); (3) MLPL (CON supplemented with 0.024% dietary LPL, DM basis); and (4) HLPL (CON supplemented with 0.048% dietary LPLs, DM basis). The results showed that dietary supplementation with LPLs linearly increased the average daily gain (p < 0.01), digestibility of DM (p < 0.01), crude protein (p < 0.01), and ether extract (p < 0.01) and decreased the feed conversion ratio (p < 0.01). A linear increase in N retention (p = 0.01) and a decrease in urinary (p = 0.04) and fecal N (p = 0.02) levels were observed with increasing the supplemental doses of LPLs. Bulls fed LPLs showed a linear increase in glutathione peroxidase (p = 0.04) and hepatic lipase (p < 0.01) activity and a decrease in cholesterol (p < 0.01), triglyceride (p < 0.01), and malondialdehyde (p < 0.01) levels. In conclusion, supplementation with LPLs has the potential to improve the growth performance, nutrient digestibility, and antioxidant status of beef cattle.

An experiment was conducted to investigate the influences of supplemental lysophospholipids (LPL) on the growth performance, nutrient digestibility, and fecal bacterial profile, and short-chain fatty acids (SCFAs) of beef cattle. Thirty-six Angus beef cattle [565 ± 10.25 kg body weight (BW)] were grouped by BW and age, and randomly allocated to 1 of 3 treatment groups: (1) control (CON, basal diet); (2) LLPL [CON supplemented with 0.5 g/kg LPL, dry matter (DM) basis]; and (3) HLPL (CON supplemented with 0.75 g/kg, DM basis). The Angus cattle were fed a total mixed ration that consisted of 25% roughage and 75% concentrate (dry matter [DM] basis). The results reveal that LPL inclusion linearly increased the average daily gain ( P = 0.02) and the feed efficiency (ADG/feed intake, P = 0.02), while quadratically increasing the final weight ( P = 0.02) of the beef cattle. Compared with CON, the total tract digestibilities of DM ( P < 0.01), ether extract ( P = 0.04) and crude protein ( P < 0.01) were increased with LPL supplementation. At the phylum-level, the relative abundance of Firmicutes ( P = 0.05) and ratio of Firmicutes : Bacteroidetes ( P = 0.04) were linearly increased, while the relative abundances of Bacteroidetes ( P = 0.04) and Proteobacteria ( P < 0.01) were linearly decreased with increasing LPL inclusion. At the genus-level, the relative abundances of Clostridium ( P < 0.01) and Roseburia ( P < 0.01) were quadratically increased, and the relative abundances of Ruminococcus was linearly increased ( P < 0.01) with LPL supplementation. Additionally, increasing the dose of LPL in diets linearly increased the molar proportion of butyrate ( P < 0.01) and total SCFAs ( P = 0.01) concentrations. A conclusion was drawn that, as a promising feed additive, LPL promoted growth performance and nutrient digestibility, which may be associated with the change of fecal microbiome and SCFAs., (Copyright © 2022 Zhang, Bai, Zhao, Wang, Li, Zhang and Jiao.)

Clostridium butyricum ( C. butyricum ) is currently widely used to improve the body health and productive performance of monogastric animals. However, there have been few reports on the effects and specific mechanism of action of Clostridium butyricum in ruminants. This study aimed to investigate the effects of Clostridium butyricum supplementation on the growth performance and digestive microbiota of fattening goats. Twenty-four healthy male Albas goats (body weight = 22 ± 2.03 kg) were randomly divided into 3 treatment groups with eight goats in each group. The treatments were as follows: control group (CON) (basal diet, concentrate to forage ratio = 65:35); low-dose Clostridium butyricum (LCB) (basal diet plus 2.0 × 10 8 CFU/kg Clostridium butyricum ); and high-dose Clostridium butyricum (HCB) (basal diet plus 1.0 × 10 9 CFU/kg Clostridium butyricum ). The experiment lasted for 8 weeks after a 2-week adaptation period. Therefore, growth performance and rumen and rectum microbiota were evaluated in goats supplemented with Clostridium butyricum and its metabolites. The results showed that dietary supplementation with Clostridium butyricum significantly increased the pH ( P < 0.05), but had no significant effect on growth performance ( P > 0.05). Compared with the control group, dietary Clostridium butyricum supplementation significantly increased the relative abundance of Prevotella&#95;1, Christensenellaceae AE&#95;R-7&#95;Group and Prevotellaceae AE&#95;UCG-003 ( P < 0.05), and significantly decreased Succiniclasticum and Muribaculaceae&#95;unclassified ( P < 0.05). The relative abundance of Clostridium in the rumen was <1.0%. Moreover, 16S rDNA analysis showed that the fecal Clostridium or Clostridium butyricum count was significantly decreased ( P < 0.05), and the relative abundance of Alistipes and Akkermansia was increased ( P < 0.10) in the low-dose group compared with the control group. Supplementing Clostridium butyricum in a high-concentrate diet did not significantly affect the performance of goats, while regulation of the gastrointestinal microbiota and related metabolites was associated with rumen fermentation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zhang, Yu, Wang, Yu, Zhang and Sun.)

Experiments were conducted to evaluate the effects of an aflatoxin B1 (AFB1)-contaminated diet treated with ammonia on the diet detoxification and growth performance, nutrient digestibility, nitrogen utilization, and blood metabolites in sheep. Twenty-four female mutton sheep with an initial body weight of 50 ± 2.5 kg were randomly assigned to one of three groups: (1) control diet (C); (2) aflatoxin diet (T; control diet supplemented with 75 μg of AFB1/kg of dry matter); and (3) ammoniated diet (AT; ammoniated aflatoxin diet). The results showed decreases (p < 0.05) in average daily feed intake, nutrient digestibility of dry matter, crude protein and ether extract, and retained nitrogen, and an increase (p < 0.05) in urine nitrogen excretion in sheep fed diet T compared with those fed the other diets. In comparison to C and AT, feeding T decreased (p < 0.05) the concentrations of total protein, immunoglobulin A, immunoglobulin G, immunoglobulin M, superoxide dismutase, and total antioxidants and increased (p < 0.05) the concentrations of alanine amino transferase, malondialdehyde, and interleukin-6. In summary, ammonia treatment has the potential to decrease the concentration of AFB1 and alleviate the adverse effects of AFB1.

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.

Fatty acid composition is a key factor affecting the flavor and quality of goat milk. CircRNAs are now recognized as important regulators of transcription, and they play an important role in the control of fatty acid synthesis. Thus, understanding the regulatory mechanisms controlling this process in ruminant mammary glands is of great significance. In the present study, mammary tissue from dairy goats during early lactation and the dry period (nonlactating) were collected and used for high-throughput sequencing. Compared to levels during the dry period, the expression level of circ003429 during early lactation was lower (12.68-fold downregulated). In isolated goat mammary epithelial cells, circ003429 inhibited the synthesis of triglycerides (TAG) and decreased the content of unsaturated fatty acids (C16:1, C18:1, and C18:2), indicating that this circRNA plays an important role in regulating lipid synthesis. A binding site for miR-199a-3p in the circ003429 sequence was detected, and a dual-luciferase reporter system revealed that circ003429 targets miR-199a-3p. Overexpression of circ003429 (pcDNA-circ003429) downregulated the abundance of miR-199a-3p. In contrast, overexpression of miR-199a-3p increased TAG content and decreased mRNA abundance of Yes-associated protein 1 ( YAP1 ) (a target gene of miR-199a-3p), and TAG content was decreased and mRNA abundance was increased in response to overexpression of circ003429. These results indicate that circ003429 alleviates the inhibitory effect of miR-199a-3p on the mRNA abundance of YAP1 by binding miR-199a-3p, resulting in subsequent regulation of the synthesis of TAG and unsaturated fatty acids.

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.

In China, the use of antibiotics growth promoters as feed additives has been banned. The goal of raising dairy heifers is to gain a relatively high body weight on a high-fiber diet at first mating or calving, thus increasing economic benefits. The objective of this experiment was to explore the effects of supplemental Clostridium butyricum ( C. butyricum ) on growth performance, rumen fermentation and microbiota, and blood parameters in Holstein heifers. Twenty Holstein heifers [mean ± standard deviation (SD); age = 182 ± 4.20 d, body weight = 197.53 ± 5.94 kg, dry matter intake (DMI) = 6.10 ± 0.38 kg] were randomly assigned to one of two diets group for a 42-day feeding period: (1) basal diet (an untreated control group, i.e., the CON group) or (2) basal diet plus daily 2 × 10 8 (colony-forming unit, CFU) of C. butyricum per kg of DMI per heifer (the CB group). The results demonstrated that C. butyricum supplementation increased the average daily gain from d 21 to 42 and DMI compared to the control group. Supplementation with C. butyricum significantly decreased the molar proportion of acetate and the acetate to propionate ratio but increased the molar proportion of butyrate and propionate. Compared with the control group, the relative abundance of Butyrivibrio fibrisolvens, Ruminococcus albus, Ruminobacter amylophilus, Ruminococcus flavefaciens , and S treptococcus bovis increased during the trial period in the CB group. However, C. butyricum had no significant effect on the blood parameters in Holstein heifers. In conclusion, these results show that feeding C. butyricum can improve growth performance and rumen fermentation without any negative impact on blood parameters in Holstein heifers., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Li, Wang, Lv, Dou and Zhang.)

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.

The purpose of this experiment was to investigate the changes of carbohydrate composition in fermented total mixed diet and its effects on rumen fermentation, methane production, and rumen microbiome in vitro. The concentrate-to-forage ratio of the total mixed ration (TMR) was 4:6, and TMR was ensiled with lactic acid bacteria and fibrolytic enzymes. The results showed that different TMRs had different carbohydrate compositions and subfractions, fermentation characteristics, and bacterial community diversity. After fermentation, the fermented total mixed ration (FTMR) group had lower contents of neutral detergent fiber, acid detergent fiber, starch, non-fibrous carbohydrates, and carbohydrates. In addition, lactic acid content and relative abundance of Lactobacillus in the FTMR group were higher. Compared with the TMR group, the in vitro ammonia nitrogen and total volatile fatty acid concentrations and the molar proportion of propionate and butyrate were increased in the FTMR group. However, the ruminal pH, molar proportion of acetate, and methane production were significantly decreased in the FTMR group. Notably, we found that the relative abundance of ruminal bacteria was higher in FTMR than in TMR samples, including Prevotella , Coprococcus , and Oscillospira . At the same time, we found that the diversity of methanogens in the FTMR group was lower than that in the TMR group. The relative abundance of Methanobrevibacter significantly decreased, while the relative abundances of Methanoplanus and vadinCA11 increased. The relative abundances of Entodinium and Pichia significantly decreased in the FTMR group compared with the TMR group. These results suggest that FTMR can be used as an environmentally cleaner technology in animal farming due to its ability to improve ruminal fermentation, modulate the rumen microbiome, and reduce methane emissions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Li, Lv, Wang, Zhou, Zhang, Wei, Sun, Lan, Dou and Zhang.)

The objectives of this study were to evaluate changes in profiles of odd- and branched-chain fatty acids (OBCFA), including pentadecanoic acid (C15:0), 13-methyltetradecanoic acid ( iso -C15:0), 12-methyltetradecanoic acid ( anteiso -C15:0), 14-methylpentadecanoic acid ( iso -C16:0), heptadecanoic acid (C17:0), 15-methylhexadecanoic acid ( iso -C17:0), and 14-methylhexadecanoic acid ( anteiso -C17:0) during in vitro fermentation of pure carbohydrates mixtures in the buffer-rumen fluid. The second objective was to correlate the changes in the OBCFA profile to the corresponding changes in ruminal fermentation parameters, microbial crude protein (MCP) synthesis, and bacterial populations. Five pure carbohydrates mixtures containing different cellulose: starch (C:S) ratios, i.e., 0:100, 25:75, 50:50, 75:25, and 100:0, were incubated for 6, 12, 18, and 24 h in vitro . The results showed that there was significant interaction ( P < 0.05) between C:S and incubation time for changes in all OBCFA profiles, except iso-C17:0. The highest concentration of total OBCFA (3.94 mg/g dry matter; DM) was observed in the residues after 24 h of fermentation when the C:S was 0:100, while the lowest concentration of OBCFA (1.65 mg/g DM) was produced after 6 h of incubation when the C:S was 50:50. The correlation analysis revealed that the concentration of iso-C16:0 might be a potential marker for the estimation of total volatile fatty acids (ρ = 0.78) and MCP synthesis (ρ = 0.82) in the rumen. Compared to starch degrading bacteria, cellulolytic bacteria had stronger correlations with OBCFA concentrations, and the strongest correlation was found between the population of Ruminococcus flavefaciens with C15:0 concentration (ρ = 0.70). Notably, this is the first paper reporting relationship between OBCFA with rumen fermentation products and microbial protein synthesis based on fermentation of pure carbohydrates mixtures in vitro , and thus avoid confounding interference from dietary protein and fat presence in the in vivo studies. However, more in-depth experiments are needed to substantiate the current findings., Competing Interests: XLiu and SZ were employed by the company Wellhope Feed Company Limited. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Xin, Khan, Liu, Jiang, Sun, Zhang, Sun, Zhang and Li.)

Researchers have confirmed the microRNA (miRNA/miR)‑epilepsy association in rodent models of human epilepsy via a comprehensive database. However, the mechanisms of miR‑142 in epilepsy have not been extensively studied. In the present study, a rat model of epilepsy was first established by an injection of lithium chloride‑pilocarpine and the successful establishment of the model was verified via electroencephalogram monitoring. The levels of miR‑142, phosphatase and tensin homolog deleted on chromosome 10 (PTEN)‑induced putative kinase 1 (PINK1), marker proteins of mitochondrial autophagy, and apoptosis‑related proteins were measured. Additionally, the pathological changes in the hippocampus, the ultrastructure of the mitochondria, and degeneration and the apoptosis of neurons were observed using different staining methods. The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the hippocampus, mitochondrial membrane potential (MTP) and reactive oxygen species (ROS) generation were detected. Furthermore, the targeting association between miR‑142 and PINK1 was predicted and verified. Consequently, apoptosis increased, and mitochondrial autophagy decreased, in the hippocampus of epileptic rats. Following miR‑142 inhibition, the epileptic rats exhibited an increased Bax expression, a decreased Bcl‑2 expression, upregulated marker protein levels of mitochondrial autophagy, a reduced MDA content, an enhanced SOD activity, an increased MTP and decreased ROS generation. PINK1 is a target gene of miR‑142, and its overexpression protected against hippocampal damage. Taken together, the results of the present study demonstrated that miR‑142 inhibition promotes mitochondrial autophagy and reduces hippocampal damage in epileptic rats by targeting PINK1. These findings may provide useful information for the treatment of epilepsy.

Background: Diacylglycerols as a fat substitute in meat products is a growing area of interest. This study was conducted to analyze the digestion rate, digestion extent, and changes in interfacial properties of lard, glycerolized lard (GL), and purified GL (PGL) in an emulsions system by pH-stat in vitro digestion., Results: PGL had significantly higher hydrolysis rate and final digestion extent than lard (P ≤ 0.05) during in vitro digestion. The analysis on diameter variations of lard, GL, and PGL during digestion indicated that the surface- and volume-weighted mean particle diameters of all samples had the same variation trend, but variation degree was different. Concurrently, the ζ-potential analysis of the lard, GL, and PGL during the digestion process showed that the absolute values of the ζ-potentials of the three types of lipids increased at first and subsequently decreased. The microstructure changes results for lard, GL, and PGL showed there was no obvious flocculation, and the particle size of lard throughout the digestion process was larger than that of GL and PGL., Conclusion: This study showed that lard-based diacylglycerols had high digestibility characteristics and could be applied as a functional lipid in meat products to improve human health. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

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.

Background: As a major disease affecting dairy cow production worldwide, bovine mastitis is caused by a variety of pathogenic microorganisms that eventually cause mammary gland inflammation. Acremonium terricola culture (ATC) is a new type of affordable feed additive produced by the solid fermentation of A. terricola isolated from Cordyceps gunnii and exerted its anti-inflammatory effect., Objectives: To evaluate the protective effects of ATC on mastitis and investigate its active mechanism, a lipopolysaccharide (LPS)-induced rat mastitis model was used in two experiments., Design: In Experiment 1, a total of 40 female Sprague-Dawley rats were used to determine the optimal supplementary dose of ATC via gavage trial. In Experiment 2, we examined the effects of an optimal dose of ATC on LPS-induced mastitis in rats., Results: The results of Experiment 1 showed that administration of ATC improved growth performance and antioxidant functions in the serum and the liver, as well as immunoglobulin A, G, and M levels in rat serum, and it decreased the content of alanine aminotransferase, aspartate aminotransferase, triglyceride, cholesterol, low-density lipoprotein, and serum urea nitrogen in rat serum; a dosage of 250-1,250 mg/kg/day was shown to be high enough to be effective. The results of Experiment 2 indicated that ATC can relieve the inflammatory reaction of mammary glands in rats, and the LPS-induced expression of tumor necrosis factor-α, interleukin-1β, interleukin-6, and inducible nitric oxide synthase significantly decreased after ATC treatment. Moreover, our results demonstrated that ATC markedly enhanced the activity of antioxidase in this rat mastitis model. The results of Western blot analysis revealed that ATC could suppress the expression of toll-like receptor 4, phosphorylation of extracellular signal-regulated kinase, and activity of c-Jun N-terminal kinase in the LPS-stimulated mastitis model., Conclusion: Taken together, ATC was shown to exert its anti-inflammatory effect by blocking mitogen-activated protein kinase signaling pathways. These results demonstrate that ATC exerts anti-inflammatory and antioxidant effects in mastitis prevention., Competing Interests: The authors have not received any funding or benefits from industry or elsewhere to conduct this study., (© 2020 Yang Li et al.)

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.

Bovine mammary epithelial cells (BMECs) of high-producing dairy cows are subject to constant oxidative stress as a result of high metabolic rate and physiological adaptation to intensive farming. Moringa (Moringa oleifera) leaf has been proposed to have the antioxidant potential in scavenging free radicals due to the presence of flavonoids. In this study, we investigated the cytoprotective effects of moringa leaf flavonoids in alleviating oxidative stress in BMECs in vitro. Oxidative stress was established by exposing isolated BMECs to H 2 O 2 for 2 hr. Doses of moringa leaf flavonoids were evaluated by treating BMECs for 12 hr. The optimal concentrations of H 2 O 2 and moringa leaf flavonoids were 500 μmol/L and 1.0 mg/ml, respectively. The results showed that moringa leaf flavonoids increased the activities of superoxide dismutase, catalase, and glutathione peroxidase; and reduced malondialdehyde activity and intracellular accumulation of reactive oxygen species (ROS) in the H 2 O 2 -induced oxidative stress system. A Hoechst33258 staining assay revealed that moringa leaf flavonoids decreased the apoptosis rate in BMECs, while leaving membrane integrity and nucleolar morphology unchanged. We concluded that moringa leaf flavonoids have the antioxidant capacity to effectively reduce oxidative stress in BMECs., (© 2020 Blackwell Verlag GmbH.)

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.

Fragile X syndrome (FXS) is one of the most common forms of inherited mental retardation; it is usually associated with the transcriptional silencing of the Fmr1 gene and loss of its encoded protein, the fragile X mental retardation protein (FMRP). FMRP is an RNA-binding protein and participates in regulating the development of dendritic spines and synaptic plasticity. To uncover the possible role of microRNAs (miRNAs) in FXS and their relationship with FMRP, we used microarray analysis to investigate the miRNA expression profiles in the hippocampal tissues of Fmr1 knockout (Fmr1-KO) mice and wild type (WT) mice. A total of 75 differentially expressed miRNAs were identified, of which 58 were significantly upregulated and no miRNAs were significantly downregulated in Fmr1-KO mice. Quantitative real-time PCR (qRT-PCR) analysis was applied to validate the expression of 7 upregulated miRNAs; results indicated that the levels of only miR-449a and miR-720 were significantly upregulated. We further used bioinformatics software and databases to predict the target genes of these two miRNAs. The genes were related to dendritic spine development and synaptic plasticity; the qRT-PCR and western blotting results showed that cyclin-dependent kinase 5 (CDK5) and synaptotagmin 1 (SYT1) were differentially expressed in the Fmr1-KO mice and WT mice. In conclusion, this study evidenced diverse changes in the expression of miRNAs, and validated the miRNAs and their targeted genes in Fmr1-KO mice. Although further studies are required to better understand the function of miRNAs in FXS, the present research highlights a potential role of miRNAs in the pathogenesis of FXS., Competing Interests: None., (AJTR Copyright © 2020.)

The aim of the study was to understand the internal relationship between milk quality and lipid metabolism in cow mammary glands. A serial of studies was conducted to assess the molecular mechanism of PRL/microRNA-183/IRS1 (Insulin receptor substrate) pathway, which regulates milk fat metabolism in dairy cows. microRNA-183 (miR-183) was overexpressed and inhibited in cow mammary epithelial cells (CMECs), and its function was detected. The function of miR-183 in inhibiting milk fat metabolism was clarified by triglycerides (TAG), cholesterol and marker genes. There is a CpG island in the 5'-flanking promoter area of miR-183, which may inhibit the expression of miR-183 after methylation. Our results showed that prolactin (PRL) inhibited the expression of miR-183 by methylating the 5' terminal CpG island of miR-183. The upstream regulation of PRL on miR-183 was demonstrated, and construction of the lipid metabolism regulation network of microRNA-183 and target gene IRS1 was performed. These results reveal the molecular mechanism of PRL/miR-183/IRS1 pathway regulating milk fat metabolism in dairy cows, thus providing an experimental basis for the improvement of milk quality.

: 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.

Background: Yeast products showed beneficial effects with respect to stabilizing ruminal pH, stimulating ruminal fermentation and improving production efficiency. Batch cultures were conducted to evaluate the effects of yeast products on gas production (GP), dry matter disappearance (DMD) and fermentation characteristics of high-forage substrate. The study was a two media pH (5.8 and 6.5) × five yeasts (three live yeasts, LY: LY1, LY2, LY3; two yeast derivatives, YD: YD4, YD5) × four dosages factorial arrangement, with monensin (Mon) assigned as a positive control., Results: Greater (P < 0.01) GP, DMD, volatile fatty acid (VFA) concentration, ratio of acetate to propionate (A:P) and copy numbers of Fibrobacter succinogenes and Ruminococcus flavefaciens were observed at pH 6.5 than at pH 5.8. The GP kinetics, DMD, VFA concentration, A:P and NH 3 -N concentration differed (P < 0.05) among yeasts but varied with media pH or yeast dosages. Increasing doses of LY3 linearly increased DMD (P < 0.04) and VFA concentration (P < 0.001) at media pH 5.8. The DMD linearly (P < 0.02) increased with increased addition of YD4 (pH 6.5) and YD5 (pH 5.8) and the ratio of A:P linearly decreased (P < 0.01) with the addition of YD4 or YD5 at pH 5.8. Overall greater (P < 0.05) GP, A:P (pH 5.8) and DMD (pH 6.5) were observed with yeast products than with Mon., Conclusion: LY3 appeared to be an interesting candidate for improving rumen digestibility and fermentation efficiency, particularly at low media pH. YD4 or YD5 improved fermentation efficiency and can be potentially fed as an alternative to Mon. © 2019 Her Majesty the Queen in Right of Canada Journal of the Science of Food and Agriculture © 2019 Society of Chemical Industry., (© 2019 Her Majesty the Queen in Right of Canada Journal of the Science of Food and Agriculture © 2019 Society of Chemical Industry.)

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.

The aim of this study was to determine if feeding Lactobacillus rhamnosus GG (GG, ATCC 53013) to neonatal calves would alter their growth, health, rumen fermentation, and bacterial community composition during the preweaning stage; we hypothesized that it would. Twenty-four male Holstein calves were blocked and randomly assigned to 1 of 2 treatment groups: 1) untreated control (CON), or 2) treated with 1 × 1010 cfu/d of a GG suspension (GG). Calves received GG daily, mixed with the milk replacer in the morning feed for 6 wk. Starter and alfalfa hay intakes, as well as feces and respiratory scores, were recorded daily, while body weight and structures were measured weekly. Blood, rumen fluid, and feces samples were collected, from which relevant indicators were detected. The results showed that the administration of GG significantly increased voluntary starter intake (P = 0.023) and ADG (P = 0.035) of the calves. The fecal score (P = 0.018) was lower and the β-hydroxybutyric concentration in the plasma tended to increase (P = 0.092) in calves treated with GG. The pH of the rumen fluid in calves fed GG was lower (P = 0.007), which might be attributable to the tendency (P = 0.083) for total volatile fatty acids concentration to increase. Administration of GG significantly increased the amylase, protease activity, and the microbial protein concentrations (P = 0.043, P = 0.036, and P = 0.044, respectively) in the rumen fluids. Furthermore, GG treatment altered the dominant bacteria order and relative abundance of the bacteria families in the rumen fluids. The microbial diversity indices were significantly affected by GG administration. In conclusion, the neonatal calves fed GG before weaning increased their voluntary starter intake and growth performance, improved the rumen fermentation, and regulated the pattern to normally increase the propionate and butyrate concentrations. Administration of GG also diversified the bacterial community composition in the rumen, and regulated the balance of rumen and intestinal microorganisms. These results indicated that feeding calves GG were beneficial to the rumen development and early weaning., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)