The Effects of Non-Fiber Carbohydrate Content and Forage Type on Rumen Microbiome of Dairy Cows.

The main objective of our current study was evaluating the effects of NFC supplementation and forage type on rumen microbiota and metabolism, by comparing microbial structures and composition among samples collected from cows fed AH (alfalfa-based diet), H-NFC (CS-based diet with high NFC) and L-NFC (CS-based diet with low NFC) diets. Our results show that microbial communities were structurally different but functionally similar among groups. When compared with L-HFC, NFC increased the population of Treponema, Ruminobacter, Selenomonas and Succinimonas that were negatively correlated with ruminal NH3-N, and urea nitrogen in blood, milk and urine, as well as significantly increasing the number of genes involved in amino acid biosynthesis. However, when compared to the AH group, H-NFC showed a higher abundance of bacteria relating to starch degradation and lactate production, but a lower abundance of bacteria utilizing pectin and other soluble fibers. This may lead to a slower proliferation of lignocellulose bacteria, such as Ruminococcus, Marvinbryantia and Syntrophococcus. Lower fibrolytic capacity in the rumen may reduce rumen rotation rate and may limit dry matter intake and milk yield in cows fed H-NFC. The enzyme activity assays further confirmed that cellulase and xylanase activity in AH were significantly higher than H-NFC. In addition, the lower cobalt content in Gramineae plants compared to legumes, might have led to the significantly down-regulated microbial genes involved in vitamin B12 biosynthesis in H-NFC compared to AH. A lower dietary supply with vitamin B12 may restrict the synthesis of milk lactose, one of the key factors influencing milk yield. In conclusion, supplementation of a CS-based diet with additional NFC was beneficial for nitrogen conversion by increasing the activity of amino acid biosynthesis in rumen microbiota in dairy cattle. However, lower levels of fibrolytic capacity may limit dry matter intake of cows fed H-NFC and may prevent increased milk yield.

Effects of Dietary Rumen-Protected Betaine Supplementation on Performance of Postpartum Dairy Cows and Immunity of Newborn Calves.

The objective of this study was to evaluate the effects of rumen-protected betaine supplementation on performance of postpartum dairy cows and immunity of newborn calves. Twenty-four multiparous Holstein dairy cows were randomly divided into the control (CON, n = 12) and rumen-protected betaine (BET, n = 12) groups after blocking by parity and milk yield during the previous lactation cycle. The cows were fed a basal total mixed ration diet without BET (CON) or with BET at 20 g/d per cow (BET) from four weeks before expected calving to six weeks postpartum. The results showed that betaine supplementation had no effect on dry matter intake and milk yield of the cows. The BET cows tended to increase feed efficiency (energy-corrected milk/dry matter intake) and body weight loss postpartum compared to the CON cows. The plasma ß-hydroxybutyrate concentrations of the BET cows were greater at d seven after calving than those of the CON cows. Moreover, compared to the CON calves, the BET calves had greater plasma total protein and globulin concentrations. The plasma glucose concentrations of the BET calves tended to decrease relative to CON cows. In conclusion, rumen-protected betaine supplementation from four weeks before expected calving tended to increase fat mobilization of postpartum dairy cows, and might improve the immunity of newborn calves.

Lactation performance and rumen fermentation in dairy cows fed a diet with alfalfa hay replaced by corn stover and supplemented with molasses.

OBJECTIVE: The objective of current study was to investigate the lactation performance and rumen fermentation characteristics of dairy cows fed a diet with alfalfa hay replaced by corn stover but supplemented with molasses. METHODS: Sixteen Holstein cows in mid-lactation were randomly assigned to 1 of 2 dietary treatments: i) alfalfa based diet (AH), and ii) corn stover based diet supplemented with molasses (CSM). The experiment was conducted according to a 2×2 crossover design with 22-d each period, consisting of 17 d for adaptation and 5 d for data and samples collection. RESULTS: Dry matter intake and milk yield were higher for cows fed AH than CSM (p<0.01). Milk protein content and nitrogen conversion were higher (p<0.05), while milk urea nitrogen was lower (p<0.01) for cows fed AH than CSM-fed cows. Contents of milk total solids, fat and lactose were not different between two groups (p>0.10). Total rumen volatile fatty acid concentration tended to be higher (p = 0.06) for cows fed AH than CSM-fed cows. Molar proportion of acetate was lower (p = 0.04), but valerate was higher (p = 0.02) in cows fed AH than CSM-fed cows. Rumen concentration of propionate, and isobutyrate, and ratio of acetate to propionate tended to be different (p<0.10) between two groups. The feed cost per kilogram of milk was lower in CSM than AH (p<0.01). No differences were found in feed efficiency and most plasma parameters tested (p>0.10). CONCLUSION: In comparison with AH diet, CSM diet could be fed to dairy cows without negative effect on feed efficiency, ruminal fermentation, but economically beneficial, indicating that CSM could be an alternative choice for dairy farms instead of AH to feed mid-lactation dairy cows.

Effects of the dietary nonfiber carbohydrate content on lactation performance, rumen fermentation, and nitrogen utilization in mid-lactation dairy cows receiving corn stover.

BACKGROUND: Corn stover (CS) is an abundant source of feed for livestock in China. However, it is low in nutritional value that we have been seeking technologies to improve. Previous studies show that non-fiber carbohydrate (NFC) might limit the utilization of a CS diet by lactating dairy cows. Thus, this study was conducted to investigate the lactation performance and rumen fermentation characteristics in lactating cows consuming CS with two contents of NFC compared to an alfalfa hay-containing diet. Twelve Holstein cows were used in a replicated 3 × 3 Latin square design with three dietary treatments: (1) low-NFC diet (NFC = 35.6%, L-NFC), (2) high-NFC diet (NFC = 40.1%, H-NFC), and (3) alfalfa hay diet (NFC = 38.9%, AH). RESULTS: Intake of DM was lower for cows fed H-NFC compared to L-NFC and AH, while the milk yield was higher in AH than in H-NFC and L-NFC (P < 0.01). The feed efficiency (milk yield/DM intake, 1.15 vs. 1.08, P < 0.01) were greater for cows fed H-NFC than L-NFC. The contents of milk protein and lactose were not different among the groups (P > 0.11), but milk fat content was higher for cows fed H-NFC and L-NFC compared to AH (P < 0.01). The rumen ammonia nitrogen concentration and the concentrations of urea nitrogen in blood and milk were lower for cows fed H-NFC and AH compared to L-NFC (P < 0.05). The concentrations of rumen propionate and total volatile fatty acids were different among groups (P < 0.05) with higher concentration for cows fed AH compared to H-NFC and L-NFC, and acetate concentration tended to be different among groups (P = 0.06). CONCLUSIONS: From the results obtained in this study, it was inferred that the increased NFC content in a diet containing corn stover can improve the feed efficiency and benefit the nitrogen conversion.

Lactation-related metabolic mechanism investigated based on mammary gland metabolomics and 4 biofluids' metabolomics relationships in dairy cows.

BACKGROUND: Lactation is extremely important for dairy cows; however, the understanding of the underlying metabolic mechanisms is very limited. This study was conducted to investigate the inherent metabolic patterns during lactation using the overall biofluid metabolomics and the metabolic differences from non-lactation periods, as determined using partial tissue-metabolomics. We analyzed the metabolomic profiles of four biofluids (rumen fluid, serum, milk and urine) and their relationships in six mid-lactation Holstein cows and compared their mammary gland (MG) metabolomic profiles with those of six non-lactating cows by using gas chromatography-time of flight/mass spectrometry. RESULTS: In total, 33 metabolites were shared among the four biofluids, and 274 metabolites were identified in the MG tissues. The sub-clusters of the hierarchical clustering analysis revealed that the rumen fluid and serum metabolomics profiles were grouped together and highly correlated but were separate from those for milk. Urine had the most different profile compared to the other three biofluids. Creatine was identified as the most different metabolite among the four biofluids (VIP = 1.537). Five metabolic pathways, including gluconeogenesis, pyruvate metabolism, the tricarboxylic acid cycle (TCA cycle), glycerolipid metabolism, and aspartate metabolism, showed the most functional enrichment among the four biofluids (false discovery rate < 0.05, fold enrichment >2). Clear discriminations were observed in the MG metabolomics profiles between the lactating and non-lactating cows, with 54 metabolites having a significantly higher abundance (P < 0.05, VIP > 1) in the lactation group. Lactobionic acid, citric acid, orotic acid and oxamide were extracted by the S-plot as potential biomarkers of the metabolic difference between lactation and non-lactation. The TCA cycle, glyoxylate and dicarboxylate metabolism, glutamate metabolism and glycine metabolism were determined to be pathways that were significantly impacted (P < 0.01, impact value >0.1) in the lactation group. Among them, the TCA cycle was the most up-regulated pathway (P < 0.0001), with 7 of the 10 related metabolites increased in the MG tissues of the lactating cows. CONCLUSIONS: The overall biofluid and MG tissue metabolic mechanisms in the lactating cows were interpreted in this study. Our findings are the first to provide an integrated insight and a better understanding of the metabolic mechanism of lactation, which is beneficial for developing regulated strategies to improve the metabolic status of lactating dairy cows.

Effects of Saccharomyces cerevisiae fermentation products on performance and rumen fermentation and microbiota in dairy cows fed a diet containing low quality forage.

BACKGROUND: A possible option to meet the increased demand of forage for dairy industry is to use the agricultural by-products, such as corn stover. However, nutritional value of crop residues is low and we have been seeking technologies to improve the value. A feeding trial was performed to evaluate the effects of four levels of Saccharomyces cerevisiae fermentation product (SCFP; Original XP; Diamond V) on lactation performance and rumen fermentation in mid-lactation Holstein dairy cows fed a diet containing low-quality forage. Eighty dairy cows were randomly assigned into one of four treatments: basal diet supplemented with 0, 60, 120, or 180 g/d of SCFP per head mixed with 180, 120, 60, or 0 g of corn meal, respectively. The experiment lasted for 10 wks, with the first 2 weeks for adaptation. RESULTS: Dry matter intake was found to be similar (P > 0.05) among the treatments. There was an increasing trend in milk production (linear, P ≤ 0.10) with the increasing level of SCFP supplementation, with no effects on contents of milk components (P > 0.05). Supplementation of SCFP linearly increased (P < 0.05) the N conversion, without affecting rumen pH and ammonia-N (P > 0.05). Increasing level of SCFP linearly increased (P < 0.05) concentrations of ruminal total volatile fatty acids, acetate, propionate, and butyrate, with no difference in molar proportion of individual acids (P > 0.05). The population of fungi and certain cellulolytic bacteria (Ruminococcus albus, R. flavefaciens and Fibrobacter succinogenes) increased linearly (P < 0.05) but those of lactate-utilizing (Selenomonas ruminantium and Megasphaera elsdenii) and lactate-producing bacteria (Streptococcus bovis) decreased linearly (P ≤ 0.01) with increasing level of SCFP. The urinary purine derivatives increased linearly (P < 0.05) in response to SCFP supplementation, indicating that SCFP supplementation may benefit for microbial protein synthesis in the rumen. CONCLUSIONS: The SCFP supplementation was effective in maintaining milk persistency of mid-lactation cows receiving diets containing low-quality forage. The beneficial effect of SCFP could be attributed to improved rumen function; 1) microbial population shift toward greater rumen fermentation efficiency indicated by higher rumen fungi and cellulolytic bacteria and lower lactate producing bacteria, and 2) rumen microbial fermentation toward greater supply of energy and protein indicated by greater ruminal VFA concentration and increased N conversion. Effects of SCFP were dose-depended and greater effects being observed with higher levels of supplementation and the effect was more noticeable during the high THI environment.

Comparative studies of two methods for miRNA isolation from milk whey.

MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS(®) followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS(®) followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (<200 nt) from milk whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100).