Effects of varying dietary forage particle size in two concentrate levels on chewing activity, ruminal mat characteristics, and passage in dairy cows

The objective of this study was to investigate the effects of varying dietary forage particle size on chewing activity, ruminal mat characteristics, passage, and in situ ruminal and total tract digestion in dairy cows at a low- and high-concentrate inclusion. The experiment was designed as a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Four ruminally cannulated late-lactating dairy cows were restrictively fed (17 kg of dry matter/d), in four 23-d periods, 1 of 4 different diets varying in the theoretical particle size (6 and 30 mm) of hay (56.6% NDF of dry matter) and in the levels (approximately 20 and 60%, dry matter basis) of a cereal-based concentrate. Ingredients of the ration were offered separately to the cows; dietary hay and low-level concentrate were offered twice daily at 0800 and 1600 h, whereas concentrate of the high-level treatment was offered in 4 meals a day at 0800, 1200, 1600, and 1900 h. This study showed that altering the forage particle size from 6 to 30 mm in a low-concentrate diet significantly increased the rumination time and ruminal mat consistency without affecting ruminal fermentation and passage. Further, particle breakdown and proportion of mat in the rumen increased, and in situ hay dry matter degradability improved, which in turn indicated a higher capacity of ruminal digesta to degrade fiber. On the other hand, increasing the forage particle size in a diet containing a high amount of concentrate increased the proportion of dry matter retained on a 1.18-mm screen from 37.5 to 42.0% and extended the rumination time by 100 min/d, as well as increasing the ruminal mat consistency. However, ruminal particle breakdown, short-term ruminal pH, fibrolytic capacity of the digesta, and proportion of mat in the rumen decreased. This was also reflected in a higher bailable liquid pool, increased fractional passage rate of solid digesta from the reticulorumen, and increased retention time in the hindgut, which in turn indicated a shift of fiber digestion from the rumen to the lower digestive tract. This study showed that the response of chewing or ruminating activity alone seemed to be insufficient to assess the dietary physical effectiveness or fiber adequacy in limit-fed dairy cows when high-concentrate diets were fed separately. Based on the results of this study, we concluded that inclusion of coarsely chopped hay in the high-concentrate diet did not appear to further improve rumen conditions and digestion when the rations were formulated to exceed the fiber requirements in limit-fed dairy cows.     

Interaction of molasses and monensin in alfalfa hay- or corn silage-based diets on rumen fermentation, total tract digestibility, and milk production by Holstein cows

Sugar supplementation can stimulate rumen microbial growth and possibly fiber digestibility; however, excess ruminal carbohydrate availability relative to rumen-degradable protein (RDP) can promote energy spilling by microbes, decrease rumen pH, or depress fiber digestibility. Both RDP supply and rumen pH might be altered by forage source and monensin. Therefore, the objective of this study was to evaluate interactions of a sugar source (molasses) with monensin and 2 forage sources on rumen fermentation, total tract digestibility, and production and fatty acid composition of milk. Seven ruminally cannulated lactating Holstein cows were used in a 5 × 7 incomplete Latin square design with five 28-d periods. Four corn silage diets consisted of 1) control (C), 2) 2.6% molasses (M), 3) 2.6% molasses plus 0.45% urea (MU), or 4) 2.6% molasses plus 0.45% urea plus monensin sodium (Rumensin, at the intermediate dosage from the label, 16 g/909 kg of dry matter; MUR). Three chopped alfalfa hay diets consisted of 1) control (C), 2) 2.6% molasses (M), or 3) 2.6% molasses plus Rumensin (MR). Urea was added to corn silage diets to provide RDP comparable to alfalfa hay diets with no urea. Corn silage C and M diets were balanced to have 16.2% crude protein; and the remaining diets, 17.2% crude protein. Dry matter intake was not affected by treatment, but there was a trend for lower milk production in alfalfa hay diets compared with corn silage diets. Despite increased total volatile fatty acid and acetate concentrations in the rumen, total tract organic matter digestibility was lower for alfalfa hay-fed cows. Rumensin did not affect volatile fatty acid concentrations but decreased milk fat from 3.22 to 2.72% in corn silage diets but less in alfalfa hay diets. Medium-chain milk fatty acids (% of total fat) were lower for alfalfa hay compared with corn silage diets, and short-chain milk fatty acids tended to decrease when Rumensin was added. In whole rumen contents, concentrations of trans-10, cis-12 C_18:2 were increased when cows were fed corn silage diets. Rumensin had no effect on conjugated linoleic acid isomers in either milk or rumen contents but tended to increase the concentration of trans-10 C_18:1 in rumen samples. Molasses with urea increased ruminal NH₃-N and milk urea N when cows were fed corn silage diets (6.8 vs. 11.3 and 7.6 vs. 12.0 mg/dL for M vs. MU, respectively). Based on ruminal fermentation characteristics and fatty acid isomers in milk, molasses did not appear to promote ruminal acidosis or milk fat depression. However, combinations of Rumensin with corn silage-based diets already containing molasses and with a relatively high nonfiber carbohydrate:forage neutral detergent fiber ratio influenced biohydrogenation characteristics that are indicators of increased risk for milk fat depression.     

Effects of yeast culture supplementation on lactation performance and rumen fermentation profile and microbial abundance in mid-lactation Holstein dairy cows

The continuous trend for a narrowing margin between feed cost and milk prices across dairy farms in the United States highlights the need to improve and maintain feed efficiency. Yeast culture products are alternative supplements that have been evaluated in terms of milk performance and feed efficiency; however, less is known about their potential effects on altering rumen microbial populations and consequently rumen fermentation. Therefore, the objective of this study was to evaluate the effect of yeast culture supplementation on lactation performance, rumen fermentation profile, and abundance of major species of ruminal bacteria in lactating dairy cows. Forty mid-lactation Holstein dairy cows (121 ± 43 days in milk; mean ± standard deviation; 32 multiparous and 8 primiparous) were used in a randomized complete block design with a 7-d adaptation period followed by a 60-d treatment period. Cows were blocked by parity, days in milk, and previous lactation milk yield and assigned to a basal total mixed ration (TMR; 1.6 Mcal/kg of dry matter, 14.6% crude protein, 21.5% starch, and 38.4% neutral detergent fiber) plus 114 g/d of ground corn (CON; n = 20) or basal TMR plus 100 g/d of ground corn and 14 g/d of yeast culture (YC; n = 20; Culture Classic HD, Cellerate Yeast Solutions, Phibro Animal Health Corp.). Treatments were top-dressed over the TMR once a day. Cows were individually fed 1 × /d throughout the trial. Blood and rumen fluid samples were collected in a subset of cows (n = 10/treatment) at 0, 30, and 60 d of the treatment period. Rumen fluid sampled via esophageal tubing was analyzed for ammonia-N, volatile fatty acids (VFA), and ruminal bacteria populations via quantitative PCR amplification of 16S ribosomal DNA genes. Milk yield was not affected by treatment effects. Energy balance was lower in YC cows than CON, which was partially explain by the trend for lower dry matter intake as % body weight in YC cows than CON. Cows fed YC had greater overall ruminal pH and greater total VFA (mM) at 60 d of treatment period. There was a contrasting greater molar proportion of isovalerate and lower acetate proportion in YC-fed cows compared with CON cows. Although the ruminal abundance of specific fiber-digesting bacteria, including Eubacterium ruminantium and Ruminococcus flavefaciens, was increased in YC cows, others such as Fibrobacter succinogenes were decreased. The abundance of amylolytic bacteria such as Ruminobacter amylophilus and Succinimonas amylolytica were decreased in YC cows than CON. Our results indicate that the yeast culture supplementation seems to promote some specific fiber-digesting bacteria while decreasing amylolytic bacteria, which might have partially promoted more neutral rumen pH, greater total VFA, and isovalerate.     

Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage

Four Holstein cows fitted with ruminal cannulas were used in a 4 × 4 Latin square design (28-d periods) with a 2 × 2 factorial arrangement of treatments to investigate the effects of addition of a specific mixture of essential oil compounds (MEO; 0 vs. 750 mg/d) and silage source [alfalfa silage (AS) vs. corn silage (CS)] on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition. Total mixed rations containing either AS or CS as the sole forage source were balanced to be isocaloric and isonitrogenous. In general, no interactions between MEO addition and silage source were observed. Except for ruminal pH and milk lactose content, which were increased by MEO supplementation, no changes attributable to the administration of MEO were observed for feed intake, nutrient digestibility, end-products of ruminal fermentation, microbial counts, and milk performance. Dry matter intake and milk production were not affected by replacing AS with CS in the diet. However, cows fed CS-based diets produced milk with lower fat and higher protein and urea N concentrations than cows fed AS-based diets. Replacing AS with CS increased the concentration of NH₃-N and reduced the acetate-to-propionate ratio in ruminal fluid. Total viable bacteria, cellulolytic bacteria, and protozoa were not influenced by MEO supplementation, but the total viable bacteria count was higher with CS- than with AS-based diets. The apparent digestibility of crude protein did not differ between the AS and CS treatments, but digestibilities of neutral detergent fiber and acid detergent fiber were lower when cows were fed CS-based diets than when they were fed AS-based diets. Duodenal bacterial N flow, estimated using urinary purine derivatives and the amount of N retained, increased in cows fed CS-based diets compared with those fed AS-based diets. Feeding cows AS increased the milk fat contents of cis-9, trans-11 18:2 (conjugated linoleic acid) and 18:3 (n-3 fatty acid) compared with feeding cows CS. Results from this study showed limited effects of MEO supplementation on nutrient utilization, ruminal fermentation, and milk performance when cows were fed diets containing either AS or CS as the sole forage source.     

Effects of supplementing a Saccharomyces cerevisiae fermentation product during the transition period on rumen fermentation of dairy cows fed fresh diets differing in starch content

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V, Cedar Rapids, IA) during the transition period (d −28 ± 3 to 23 ± 3 relative to calving) on rumen fermentation and mRNA abundance of genes in the rumen epithelium of fresh cows (d 1 to 23 ± 3 after calving) fed diets differing in starch content. Eighteen ruminally cannulated multiparous Holstein cows were fed diets with SCFP (n = 9) or without (CON; n = 9) throughout the experiment. All cows were fed a common basal controlled-energy close-up diet (1.43 Mcal/kg, net energy for lactation; 13.8% starch) before calving. Cows within each treatment (CON or SCFP) were fed either a low-starch (LS; 22.1% starch) or high-starch (HS; 28.3% starch) diet during the fresh period. Cows were assigned to treatment after balancing for parity, body condition score, and expected calving date. Rumen pH was measured continuously for 72 h starting on d −10, −3, 1, 7, and 21 relative to calving date. Rumen papillae were collected on d −10 and 21 relative to calving. Supplementation of SCFP had no effect on rumen pH during d −10 to −8, but mean rumen pH tended to be higher (6.64 vs. 6.49) for SCFP cows than for CON cows during d −3 to −1. Feeding SCFP decreased the range of rumen pH variation compared with CON within the HS group during both d 7 to 9 (1.08 vs. 1.38) and d 21 to 23 (1.03 vs. 1.30) after calving. In addition, nadir rumen pH tended to be higher (5.64 vs. 5.44) and duration of pH below 5.8 tended to be shorter (116 vs. 323 min/d) for the SCFP group than for the CON group during d 21 to 23 after calving. Supplementation of SCFP increased the mRNA abundance of insulin-like growth factor-6 (1.10 vs. 0.69) before calving and decreased the mRNA abundance of putative anion transporter isoform 1 (1.12 vs. 2.27) after calving. Nadir rumen pH tended to be higher during d 1 to 3 (5.63 vs. 5.41) for LS cows than for HS cows, but rumen pH was not affected by dietary starch content during other time periods. Dietary starch content had no effect on mRNA abundance of genes in the rumen epithelium after calving. These results suggest that supplementation of SCFP may reduce the range of variation in rumen pH in fresh cows fed HS diets and the duration of subacute ruminal acidosis by the end of the fresh period regardless of dietary starch content and that decreasing dietary starch content during the fresh period may reduce the decrease in rumen pH immediately after parturition.     

Alterations in ruminal bacterial populations at induction and recovery from diet-induced milk fat depression in dairy cows

Ten ruminally cannulated Holstein cows were used in a crossover design that investigated changes in ruminal bacterial populations in response to induction and recovery from diet-induced milk fat depression (MFD). Further, the effect on the ruminal microbiota of the cows with diet-induced milk fat depression inoculated with rumen contents from non-milk fat-depressed donor cows was evaluated. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber, high-starch, and high-polyunsaturated fatty acid diet (26.1% neutral detergent fiber, 28.1% starch, 5.8% total fatty acids, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. Induction was followed by a recovery phase, where all cows were switched to a high-fiber, low-starch, and low-polyunsaturated fatty acid diet (31.8% neutral detergent fiber, 23% starch, 4.2% total fatty acids, and 1.2% C18:2) and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Ruminal samples were collected at the end of induction (d 10) and during recovery (d 13, 16, and 28), separated to solid and liquid fractions, extracted for DNA, PCR- amplified for the V1-V2 region of the 16S rRNA gene, and analyzed for bacterial diversity. Results indicated that bacterial communities were different between fractions. In each fraction, differences were significant between the induction (d 10) and recovery (d 13, 16, and 28) periods; however, differences were less apparent with time during the recovery period. The MFD (d 10) was typified by a reduction in the relative sequence abundance of Bacteroidetes and an increase in the relative sequence abundance of Firmicutes and Actinobacteria across both fractions. At the genus level, relative sequence abundance of unclassified Lachnospiraceae, Butyrivibrio, Bulleidia, and Coriobacteriaceae were higher on d 10 and were positively correlated with trans-10,cis-12 CLA and the trans-10 isomer, suggesting their potential role in altered biohydrogenation reactions. A switch to the recovery diet resulted in a sharp increase in the Bacteroidetes lineages and a decrease in Firmicutes members on d 13; however, this shift appears to stabilize by d 28, indicating the restoration process for ruminal bacteria from an altered state is gradual and complex. Inoculation of 10% of rumen contents from non-MFD donor cows to MFD cows revealed this procedure had transient effects on only a few bacterial populations, and such effects disappeared after d 16 following cessation of inoculation. It can be concluded that alterations in milk FA profiles at induction are preceded by microbial alterations in the rumen driven by dietary changes.     

Temporal changes in total and metabolically active ruminal methanogens in dairy cows supplemented with 3-nitrooxypropanol

The purpose of this study was to investigate the effect of 3-nitrooxypropanol (3-NOP), a potent methane inhibitor, on total and metabolically active methanogens in the rumen of dairy cows over the course of the day and over a 12-wk period. Rumen contents of 8 ruminally cannulated early-lactation dairy cows were sampled at 2, 6, and 10 h after feeding during wk 4, 8, and 12 of a randomized complete block design experiment in which 3-NOP was fed at 60 mg/kg of feed dry matter. Cows (4 fed the control and 4 fed the 3-NOP diet) were blocked based on their previous lactation milk yield or predicted milk yield. Rumen samples were extracted for microbial DNA (total) and microbial RNA (metabolically active), PCR amplified for the 16S rRNA gene of archaea, sequenced on an Illumina platform, and analyzed for archaea diversity. In addition, the 16S copy number and 3 ruminal methanogenic species were quantified using the real-time quantitative PCR assay. We detected a difference between DNA and RNA (cDNA)-based archaea communities, revealing that ruminal methanogens differ in their metabolic activities. Within DNA and cDNA components, methanogenic communities differed by sampling hour, week, and treatment. Overall, Methanobrevibacter was the dominant genus (94.3%) followed by Methanosphaera, with the latter genus having greater abundance in the cDNA component (14.5%) compared with total populations (5.5%). Methanosphaera was higher at 2 h after feeding, whereas Methanobrevibacter increased at 6 and 10 h in both groups, showing diurnal patterns among individual methanogenic lineages. Methanobrevibacter was reduced at wk 4, whereas Methanosphaera was reduced at wk 8 and 12 in cows supplemented with 3-NOP compared with control cows, suggesting differential responses among methanogens to 3-NOP. A reduction in Methanobrevibacter ruminantium in all 3-NOP samples from wk 8 was confirmed using real-time quantitative PCR. The relative abundance of individual methanogens was driven by a combination of dietary composition, dry matter intake, and hydrogen concentrations in the rumen. This study provides novel information on the effects of 3-NOP on individual methanogenic lineages, but further studies are needed to understand temporal dynamics and to validate the effects of 3-NOP on individual lineages of ruminal methanogens.     

A comparison of sampling sites, digesta and microbial markers, and microbial references for assessing the postruminal supply of nutrients in dairy cows

This study evaluated the impact of some methodological factors on the flows of nutrients at the omasal canal and duodenum of dairy cows fed corn-based diets. Three ruminally and duodenally cannulated cows were assigned to an incomplete 4 × 4 Latin square with four 14-d periods and fed diets formulated to contain different amounts and ruminal degradabilities of crude protein. Samples from the omasal canal and duodenum were obtained and processed according to methodologies routinely used in our laboratories and elsewhere. Methodological factors that were evaluated included microbial references and markers, digesta markers, and sampling sites (techniques). Considerable variation was found for the compositions of microbial references and their impact on the intestinal supply of microbial nonammonia nitrogen. Likewise, it appears that variation in measuring the ruminal outflow of nitrogen fractions of microbial and dietary origin could be reduced by using ¹⁵N rather than purines as microbial markers. Sampling from the omasum and duodenum resulted in differences for ruminal outflow and site of digestion as well as digestibility of some nutrients, particularly nitrogen fractions and starch. A sizable portion of this variation was associated with deviations from the assumed ideal behavior of digesta markers and collection of samples that were unrepresentative of true digesta. Collectively, outcomes from this study indicate that more research will be required to determine the accuracy of nutrient flows and the agreement between measurements at the omasal canal and duodenum when dairy cows are fed a variety of diets under different feeding systems. Therefore, caution is recommended when extrapolating or interpreting the underlying biology of published results as well as the results of their application (e.g., model parameters and predictions).     

Effects of supplementing a Saccharomyces cerevisiae fermentation product during the periparturient period on the immune response of dairy cows fed fresh diets differing in starch content

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V, Cedar Rapids, IA) during the periparturient period (d ?28 ± 3 to 44 ± 3 relative to calving) on mRNA abundance of genes in the rumen epithelium, inflammation indicators, oxidative status, and adaptive immunity of dairy cows fed diets with different starch content after calving. From d 28 ± 3 (± standard deviation) before the expected calving date to calving, Holstein cows (n = 38) received a common basal controlled-energy close-up diet (1.43 Mcal/kg, net energy for lactation; 13.8% starch) with (SCFP; n = 19) or without (CON; n = 19) SCFP, and cows within each treatment (CON or SCFP) were fed either a low- (LS; 22.1% starch) or high-starch (HS; 28.3% starch) diet from d 1 to 23 ± 3 after calving (fresh period). There were 4 treatment groups: LS + CON (n = 9), LS + SCFP (n = 10), HS + CON (n = 10), and HS + SCFP (n = 9). From d 24 ± 3 to 44 ± 3 after calving, all cows were fed the HS diets (post-fresh period). Animal assignment to treatments was balanced for parity, body condition score, and expected calving date. An interaction was observed between dietary starch content and SCFP on indices of oxidative stress; plasma concentrations of total antioxidant capacity tended to be reduced on d 21 after calving for SCFP compared with CON cows when a LS fresh diet was fed, but did not differ for cows fed HS fresh diets. Regardless of starch content, SCFP supplementation increased plasma concentrations of malondialdehyde at d 21 after calving compared with CON. Supplementing with SCFP reduced serum concentrations of haptoglobin on d 7 after calving, indicating reduced inflammation, and feeding LS fresh diets reduced mRNA abundance of IL receptor associated kinase-1 in rumen tissue at d 21 after calving, suggesting reduced immune activation in rumen tissue. Other than the anti-inflammatory effects indicated by lower serum haptoglobin concentration, no other effects of treatment on adaptive immunity were detectable. These results indicate that supplementing SCFP through the transition period and feeding low-starch diets during the fresh period may reduce inflammation.     

Effects of feeding grass or red clover silage cut at two maturity stages in dairy cows. 1. Nitrogen metabolism and supply of amino acids

This study investigated the effects of plant species (red clover vs. timothy-meadow fescue) and forage maturity at primary harvest (early vs. late cut silage) on rumen fermentation, nutrient digestion, and nitrogen metabolism including omasal canal AA flow and plasma AA concentration in lactating cows. Five dairy cows equipped with rumen cannulas were used in a study designed as a 5 × 5 Latin square with 21-d periods. The diets consisted of early-cut and late-cut grass and red clover silage, respectively, and a mixture of late-cut grass and early-cut red clover silages given ad libitum with 9 kg/d of a standard concentrate. Grass silage dry matter intake tended to decrease but that of red clover silages tended to increase with advancing maturity. Milk yields were unchanged among treatments, milk protein and fat concentrations being lower for red clover than for grass silage diets. Rumen fluid pH was unchanged but volatile fatty acid and ammonia concentrations were higher for red clover than for grass silage diets. Intake of N, and omasal canal flows of total nonammonia N (NAN), microbial NAN, and dietary NAN were higher for red clover than for grass silage diets but were not affected by forage maturity. However, microbial NAN flow and amount of N excreted in the feces decreased with advancing maturity for grass diets but increased for red clover diets. Apparent ruminal N degradability of the diets was unchanged, but true ruminal N degradability decreased and efficiency of microbial synthesis increased with red clover diets compared with grass silage diets. Omasal canal flows of AA, except those for Met and Cys, were on average 20% higher for red clover than grass silage diets. Omasal canal digesta concentrations of Leu, Phe, branched-chain, and essential AA were higher but those of Met lower for red clover than for grass silage diets. Plasma AA concentrations, except for His (unchanged) and Met (lower), were higher for red clover than for grass diets. However, none of these AA-related variables were affected by forage maturity. Total digestibility of N and excretion of N in the urine were higher for red clover than for grass diets and decreased with advancing maturity. It was concluded that despite the higher total AA supply of cows fed red clover versus grass silage diets, further milk production responses on red clover diets were compromised by an inadequate supply of Met as evidenced by lower Met concentration in the AA profile of omasal digesta and plasma.     

The effects of feeding 3-nitrooxypropanol on methane emissions and productivity of Holstein cows in mid lactation

The objective of the current study was to determine the effects of adding 3-nitrooxypropanol to the diet of lactating Holstein cows on methane emissions, rumen fermentation, ruminal microbial profile, and milk production. Twelve ruminally cannulated Holstein cows in midlactation were used in a crossover design study with 28-d periods. Cows were fed a diet containing 38% forage on a dry matter basis with either 2,500 mg/d of 3-nitrooxypropanol (fed as 25 g of 10% 3-nitrooxypropanol on silicon dioxide) or 25 g/d of silicon dioxide (control). After a 21-d diet adaptation period, dry matter intake (DMI) and milk yield were recorded daily. Rumen fluid and digesta were collected on d 22 and 28 for volatile fatty acid analysis and microbial profiling. Enteric methane emissions were measured on d 23 to 27 using the sulfur hexafluoride tracer gas technique. Feeding 3-nitrooxypropanol did not affect DMI; however, methane production was reduced from 17.8 to 7.18 g/kg of DMI. No change in milk or milk component yields was observed, but cows fed 3-nitrooxypropanol gained more body weight than control cows (1.06 vs. 0.39 kg/d). Concentrations of total volatile fatty acids in ruminal fluid were not affected by treatment, but a reduction in acetate proportion and a tendency for an increase in propionate proportion was noted. As such, a reduction in the acetate-to-propionate ratio was observed (2.02 vs. 2.36). Protozoa counts were not affected by treatment; however, a reduction in methanogen copy count number was observed when 3-nitrooxypropanol was fed (0.95 vs. 2.69 × 10⁸/g of rumen digesta). The data showed that feeding 3-nitrooxypropanol to lactating dairy cows at 2,500 mg/d can reduce methane emissions without compromising DMI or milk production.     

Evaluation of potential carryover effects associated with limit feeding of gravid Holstein heifers

Ninety-six Holstein heifers (400 ± 6 kg, 15.2 ± 0.1 mo), including 9 with ruminal cannulas, were offered 1 of 3 diets for 180 ± 8 d in a randomized replicated pen design. Dietary treatments included a control diet (C100) and 2 independent limit-fed (LF) diets. The LF diets included one offered at 85% of C100 intake (L85) without an ionophore, and a second containing an ionophore (325 mg/head per day of lasalocid) that was offered at 80% of C100 intake (L80+I). Heifers were evaluated for growth, rumen digesta volume, nutrient excretion, and subsequent lactation performance. Limit-fed heifers consumed less dry matter and neutral detergent fiber, and had greater respective average daily gains (0.96 or 0.89 vs. 0.81 kg/d), and lower feed:gain ratios (9.1 or 9.3 vs. 13.0 kg/kg) compared with heifers offered the C100 diet. No differences in rumen pH, NH₃-N, or volatile fatty acid concentrations were observed between C100 and LF heifers. Rumen digesta volume, density, and weight were unaffected by LF, and feeding L85 or L80+I did not result in carryover effects for rumen digesta volume when these heifers were offered a common high-fiber diet immediately after the 180-d growth trial. At parturition, no differences were observed for dystocia index, calf body weight, or 7-d postpartum body weight between cows offered LF or C100 diets as heifers. Lactation body weight, dry matter intake, and feed efficiency of cows did not differ between treatments at 45 or 90 d in milk. Milk yield and milk components also were not different between cows that were offered C100 or LF diets as gravid heifers. At 45 d in milk, rumen digesta volume was greater (99.1 vs. 66.1 L) for cows offered L85 compared with cows offered L80+I as gravid heifers, but this effect was not observed at 90 d in milk. Limit feeding of gravid Holstein heifers for 180 d did not result in any carryover effects during their first lactation for rumen digesta volume, dry matter intake, or milk yield.     

Nitrate improves ammonia incorporation into rumen microbial protein in lactating dairy cows fed a low-protein diet

Generation of ammonia from nitrate reduction is slower compared with urea hydrolysis and may be more efficiently incorporated into ruminal microbial protein. We hypothesized that nitrate supplementation could increase ammonia incorporation into microbial protein in the rumen compared with urea supplementation of a low-protein diet fed to lactating dairy cows. Eight multiparous Chinese Holstein dairy cows were used in a crossover design to investigate the effect of nitrate or an isonitrogenous urea inclusion in the basal low-protein diet on rumen fermentation, milk yield, and ruminal microbial community in dairy cows fed a low-protein diet in comparison with an isonitrogenous urea control. Eight lactating cows were blocked in 4 pairs according to days in milk, parity, and milk yield and allocated to urea (7.0 g urea/kg of dry matter of basal diet) or nitrate (14.6 g of NO₃^?/kg of dry matter of basal diet, supplemented as sodium nitrate) treatments, which were formulated on 75% of metabolizable protein requirements. Nitrate supplementation decreased ammonia concentration in the rumen liquids (?33.1%) and plasma (?30.6%) as well as methane emissions (?15.0%) and increased dissolved hydrogen concentration (102%), microbial N (22.8%), propionate molar percentage, milk yield, and 16S rRNA gene copies of Selenomonas ruminantium. Ruminal dissolved hydrogen was positively correlated with the molar proportion of propionate (r = 0.57), and negatively correlated with acetate-to-propionate ratio (r = ?0.57) and estimated net metabolic hydrogen production relative to total VFA produced (r = ?0.58). Nitrate reduction to ammonia redirected metabolic hydrogen away from methanogenesis, enhanced ammonia incorporation into rumen microbial protein, and shifted fermentation from acetate to propionate, along with increasing S. ruminantium 16S rRNA gene copies, likely leading to the increased milk yield.     

Assessment of ruminal bacterial populations and protozoal generation time in cows fed different methionine sources

Methionine supplemented as 2-hydroxy-4-(methylthio)-butanoic acid (HMB) has been suggested to alter bacterial or protozoal populations in the rumen. Our objective was to determine if source of Met would change microbial populations in the rumen and to compare those results to samples from the omasum. The ruminal and omasal samples were collected from cows fed control (no Met), dl-Met, HMB, or the isopropyl ester of HMB (HMBi; estimated 50% rumen protection) in a replicated 4 × 4 Latin square design. In one square, changes in protozoal populations were determined using microscopic counts and denaturing gradient gel electrophoresis (DGGE), whereas changes in bacterial populations were determined using DGGE and ribosomal intergenic spacer length polymorphism (RIS-LP). Neither the protozoal counts nor the DGGE banding patterns derived from protozoa were different among the dietary treatments or for ruminal vs. omasal samples. As revealed by both DGGE and RIS-LP, bacterial populations clustered by treatments in ruminal and especially in omasal samples. Using cows from both Latin squares, the flow of protozoal cells from the rumen was quantified by multiplying protozoal cell count in omasal fluid by the omasal fluid flow (using CoEDTA as a liquid flow marker) or was estimated by rumen pool size of cells multiplied by either the ruminal dilution rate of CoEDTA (after termination of CoEDTA dosing) or the passage rate of Yb-marked particles. Compared with the omasal fluid flow measurement (16.4 h), protozoal generation time was approximated much more closely using the particulate than the fluid passage rate from the rumen (generation times of 15.7 and 7.5 h, respectively). There seems to be minimal selective retention of protozoal genera in the rumen in dairy cattle fed every 2 h. Data support the validity of the omasal sampling technique under our conditions.     

The effect of rumen digesta inoculation on the time course of recovery from classical diet-induced milk fat depression in dairy cows

Ten ruminally cannulated cows were used in a crossover design that investigated the effect of rumen digesta inoculation from non-milk fat-depressed cows on recovery from classical diet-induced milk fat depression (MFD) characterized by reduced fat yield, reduced de novo milk fat synthesis, and increased alternate trans isomers. Two additional cows fed a high-fiber and low-polyunsaturated fatty acid (FA) diet (31.8% neutral detergent fiber, 4.2% FA, and 1.2% C18:2) were used as rumen digesta donors. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber and high-polyunsaturated FA diet (induction; 26.1% neutral detergent fiber, 5.8% FA, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. A recovery phase followed where all cows were switched to the high-forage, low-polyunsaturated FA diet and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Milk yield and composition were measured every 3 d. Milk yield progressively decreased during recovery. Milk fat concentration increased progressively during the recovery phase and no effect of treatment existed at any time point. Also, no treatment effect of milk fat yield was detected. The concentration of milk de novo FA increased progressively during recovery for both treatments and was higher for inoculated compared with control cows on d 6. In agreement, milk fat concentration of trans-10,cis-12 conjugated linoleic acid decreased progressively in both treatments and was lower in inoculated cows on d 3 and 6. Ruminal inoculation from non-milk fat-depressed cows did not change milk fat yield, but slightly accelerated the rate of recovery of de novo FA synthesis and normal ruminal FA biohydrogenation, demonstrating a possible opportunity for other interventions that improve the ruminal environment to accelerate recovery from this condition.     

Active dry Saccharomyces cerevisiae can alleviate the effect of subacute ruminal acidosis in lactating dairy cows

The objective of the study was to determine the effect of active dry Saccharomyces cerevisiae (ADSC) supplementation on dry matter intake, milk yield, milk components, ruminal pH, and microbial community during a dietary regimen that leads to subacute ruminal acidosis (SARA). Sixteen multiparous, rumen-cannulated lactating Holstein cows were randomly assigned to 1 of 2 dietary treatments that included ADSC (Biomate; AB Vista, Marlborough, UK; 8 × 10¹⁰ cfu/head per day) or control. During wk 1 to 6, all cows received a high-forage (HF) diet (77:23, forage:concentrate). Cows were then abruptly switched during wk 7 to a high-grain (HG) diet (49:51, forage:concentrate) and remained on the HG until the end of wk 10. Feed intake and milk yields were recorded daily. Ruminal pH was recorded continuously using an indwelling system for 1 to 2 d per week during the pre-experimental phase, and wk 6, 7, and 10. Ruminal digesta samples were collected at the end of the experiment and analyzed for relative change in microbial communities using real-time quantitative PCR. Cows were considered to have SARA if the duration below pH 5.6 was ≥300 min/d. Ruminal pH during wk 6 (HF plateau) was not different across treatments (15 ± 46 min/d at pH <5.6). The dietary regimen successfully induced SARA during wk 7 (transition from HF to HG diet), and ruminal pH (551 ± 46 min/d at pH <5.6) was not different across treatments. However, cows receiving ADSC had an improved ruminal pH (122 ± 57 vs. 321 ± 53 min/d at pH <5.6) during wk 10 (HG plateau) compared with control. Additionally, cows receiving ADSC had a better dry matter intake (23.3 ± 0.66 vs. 21.6 ± 0.61 kg/d) and 4% fat-corrected milk yield (29.6 ± 1.2 vs. 26.5 ± 1.2 kg/d) than control cows during the HG phase (wk 8 to 10). During HG feeding, cows receiving ADSC had greater total volatile fatty acid and propionate concentrations (175 ± 7.5 vs. 154 ± 7.5 and 117 ± 6.1 vs. 94 ± 5.7 mM for ADSC and control, respectively) and lower acetate:propionate ratio (0.26 ± 0.5 vs. 0.36 ± 0.05 for ADSC and control, respectively). Microbial analyses conducted on samples collected during wk 10 showed that cows supplemented with S. cerevisiae had a 9-fold, 2-fold, 6-fold, 1.3-fold, and 8-fold increase in S. cerevisiae, Fibrobacter succinogenes, Anaerovibrio lipolytica, Ruminococcus albus, and anaerobic fungi, respectively, which suggested an increase in cellulolytic microbes within the rumen. Cows supplemented with ADSC had 2.2-fold reduction in Prevotella albensis, which is a gram-negative bacterium predominant during SARA. Prevotella spp. are suggested to be an important source of lipopolysaccharide responsible for inflammation within the rumen. Cows supplemented with ADSC had a 2.3-fold increase in Streptococcus bovis and a 12-fold reduction in Megasphaera elsdenii. The reduction in M. elsdenii may reflect lower concentration of lactic acid within the rumen for ADSC cows. In conclusion, ADSC supplementation to dairy cows was demonstrated to alleviate the condition of SARA caused by abrupt dietary changes from HF to HG, and can potentially improve rumen function, as indicated by greater numbers of cellulolytic microorganisms within the rumen.     

Effect of dietary soybean meal and fish meal on protein digesta flow in Holstein cows during early and midlactation

Six lactating cows were fitted with ruminal and duodenal cannula to measure protein digesta flow to the duodenum during early and midlactation. Diets were composed of corn grain, corn silage, and orchardgrass hay plus supplemental fish meal or soybean meal. Diets contained 15.5% CP and 20.7 ADF. Cobalt-EDTA and Yb were used as liquid and particulate digesta markers and cytosine was used as microbial marker. Corrected organic matter digestibilities in the stomachs were 48.4, 49.8, 44.9, and 53.2% for fish meal and soybean meal diets and early and midlactation, respectively. Preduodenal degradabilities were 47.2, 65.8, 56.7, and 56.2% for fish meal and soybean meal diets and early and midlactation, respectively. Nitrogen recoveries at the duodenum were 93.2 and 84.3% for fish meal and soybean meal diets. Intake of amino acids was greater when cows were fed the soybean meal diet, but total flows of amino acids to the duodenum were similar for both diets. Greater quantity of protein escaping ruminal degradation in cows fed fish meal compared with soybean meal was counterbalanced by less microbial synthesis in the rumen.     

Digestion, Ruminal Fermentation, Ciliate Protozoal Populations, and Milk Production from Dairy Cows Fed Cinnamaldehyde, Quebracho Condensed Tannin, or Yucca schidigera Saponin Extracts

Four ruminally cannulated lactating cows were used in a 4 × 4 Latin square design (28-d periods) to determine the effects of cinnamaldehyde (CIN; 1 g/cow per day), condensed tannins from quebracho trees (QCT, containing 70% tannins, 150 g/cow per day), and saponins from Yucca schidigera extract (YSE, containing 10% saponins; 60 g/cow per day) on digestion, ruminal fermentation characteristics, protozoal populations, and milk production. Intake of dry matter was not affected by the addition of CIN or QCT, but cows fed YSE had lesser intake than cows fed the control diet (21.8 vs. 23.2 kg/d). Apparent total-tract digestibilities of dry matter, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber were unaffected by dietary treatments. Supplementation with CIN, QTE, or YSE did not affect in situ ruminal degradation of soybean meal, grass silage, or corn grain. Ruminal pH (6.67), total volatile fatty acid concentration (135 mM), and molar proportions (mol/100 mol of total volatile fatty acid) of acetate (65.0), propionate (19.6), and butyrate (11.2) were similar among treatments. Ruminal NH₃-N concentration was not changed by the addition of CIN and YSE, but tended to decrease in cows fed QCT compared with cows fed the control diet (132 vs. 160 mg/L). Total numbers of ruminal protozoa were not changed by adding CIN, QCT, or YSE in the diet (5.85 log₁₀/mL). However, the number of Isotricha was greater in ruminal fluid of cows fed CIN than in ruminal fluid of cows fed the control diet (4.46 vs. 4.23 log₁₀/mL). Milk production (33.1 kg/d), milk fat (4.3%), and milk protein (3.5%) remained unchanged between dietary treatments. Results of this study show that under our experimental conditions, supplementing dairy cow diets with CIN, QCT, or YSE had limited effects on digestion, ruminal fermentation characteristics, and protozoal populations. The lack of effects observed in this study suggests that these antimicrobials require administration at greater doses to favorably alter rumen microbial fermentation.     

Saccharomyces cerevisiae fermentation products reduce bacterial endotoxin concentrations and inflammation during grain-based subacute ruminal acidosis in lactating dairy cows

Subacute ruminal acidosis (SARA) is a metabolic disorder in dairy cows that is associated with dysbiosis of rumen and hindgut microbiomes, translocation of immunogenic compounds from the gut lumen into blood circulation, and systemic inflammatory response. In this study we hypothesized that Saccharomyces cerevisiae fermentation products (SCFP) attenuate the increases in ruminal and peripheral bacterial endotoxin concentrations and the inflammation resulting from repeated induction of SARA. Lactating Holstein dairy cows (parity 2 and 3+, n = 32) were fed diets with or without SCFP (all from Diamond V) and subjected to 2 episodes of SARA challenges. Cows received a basal total mixed ration (TMR) containing 34% neutral detergent fiber and 18.6% starch, dry matter (DM) basis. Treatments were randomly assigned to control (basal TMR and 140 g/d of ground corn with no SCFP) or 1 of 3 SCFP treatments: basal TMR and 14 g/d Original XPC (SCFPa), 19 g/d NutriTek (SCFPb-1×), or 38 g/d NutriTek (SCFPb-2×) mixed with 126, 121, or 102 g/d of ground corn, respectively. Treatments were implemented from 4 wk before until 12 wk after parturition. During wk 5 (SARA1) and wk 8 of lactation (SARA2), grain-based SARA challenges were conducted by gradually replacing 20% of DM of the basal TMR over 3 d with pellets containing 50% wheat and 50% barley. Ruminal fluid, fecal, and blood samples were collected weekly during Pre-SARA1 (wk 4, as baseline), Post-SARA1 (wk 7), and Post-SARA2 (wk 10 for blood and wk 12 for rumen and fecal parameters) stages, and twice a week during the challenges SARA1 and SARA2. Rumen papillae samples were taken only during Pre-SARA1 and Post-SARA2. We measured the concentrations of free lipopolysaccharides (LPS) in the rumen fluid and feces; free LPS and lipoteichoic acid (LTA) endotoxins in peripheral plasma; interleukin (IL)-1β and IL-6 in peripheral serum; acute-phase proteins, serum amyloid A (SAA), and LPS-binding protein in peripheral plasma; haptoglobin (Hp) in peripheral serum; and myeloperoxidase (MPO) in rumen papillae. Induction of SARA episodes increased free LPS concentrations in rumen fluid and tended to increase LTA in peripheral plasma. The SARA episodes increased concentration of circulating SAA and tended to increase that of IL-1β compared with Pre-SARA1. Induction of SARA did not affect the concentrations of circulating IL-6, Hp, and MPO. The SCFP supplementation reduced plasma concentrations of LTA and SAA and serum concentration of IL-1β compared with control. Additionally, SCFPb-2× tended to reduce ruminal LPS in second-parity cows compared with control. Overall, SCFP supplementation appeared to stabilize the rumen environment and reduce proinflammatory status, hence attenuating adverse digestive and inflammatory responses associated with SARA episodes.     

Ration formulations containing reduced-fat dried distillers grains with solubles and their effect on lactation performance,rumen fermentation,and intestinal flow of microbial nitrogen in Holstein cows

Sixteen multiparous lactating Holstein cows were used in 2 experiments to evaluate the effects of reduced-fat dried distillers grains with solubles (RFDG) on milk production, rumen fermentation, intestinal microbial N flow, and total-tract nutrient digestibility. In experiment 1, RFDG was fed at 0, 10, 20, or 30% of diet dry matter (DM) to 12 noncannulated Holstein cows (mean ± standard deviation: 89 ± 11 d in milk and 674 ± 68.2 kg of body weight) to determine effects on milk production. In experiment 2, the same diets were fed to 4 ruminally and duodenally cannulated Holstein cows (mean ± standard deviation: 112 ± 41 d in milk; 590 ± 61.14 kg of body weight) to evaluate the effects on rumen fermentation, intestinal flow of microbial N, and total-tract nutrient digestibility. In both experiments, cows were randomly assigned to 4 × 4 Latin squares over 21-d periods. Treatments (DM basis) were (1) control (0% RFDG), (2) 10% RFDG, (3) 20% RFDG, and (4) 30% RFDG. Feed intake and milk yield were recorded daily. In both experiments, milk samples were collected on d 19 to 21 of each period for analysis of milk components. In experiment 2, ruminal pH was measured; samples of rumen fluid, duodenal digesta, and feces were collected on d 18 to 21. Microbial N was estimated by using purines and DNA as microbial markers. Milk yield was not affected by treatment and averaged 34.0 ± 1.29 kg/d and 31.4 ± 2.81 kg/d in experiments 1 and 2, respectively. Percentage of milk protein tended to increase in experiment 1; estimates were 3.08, 3.18, 3.15, and 3.19 ± 0.06% when RFDG increased from 0 to 30% in the diets. However, milk protein concentration was not affected in experiment 2 and averaged 3.02 ± 0.07%. Percentage of milk fat was not affected and averaged 3.66 ± 0.05% and 3.25 ± 0.14% in experiments 1 and 2, respectively. Total ruminal volatile fatty acids and ammonia concentrations were not affected by treatment and averaged 135.18 ± 6.45 mM and 18.66 ± 2.32 mg/dL, respectively. Intestinal microbial N flow was not affected by treatment; however, purines yielded higher estimates of flow compared with DNA markers. When averaged across treatments, intestinal flow of microbial N was 303 and 218 ± 18 g of N/d, using purines and DNA as the markers. Dry matter, organic matter, neutral detergent fiber, and nonfiber carbohydrate digestibility tended to increase with increasing inclusion of RFDG. Results from these experiments indicate that dairy rations can be formulated to include up to 30% RFDG while maintaining lactation performance, volatile fatty acids concentration, and intestinal supply of microbial N.