Effect of incremental urea supplementation of a conventional corn silage-based diet on ruminal ammonia concentration and synthesis of microbial protein

One primiparous and 3 multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 × 4 Latin square design to determine the efficacy of adding urea to a corn silage-based diet on ruminal fermentation and microbial protein synthesis. Dietary treatments were 0, 0.3, 0.6, and 0.9% urea in diet dry matter (DM); urea was manually top dressed and incorporated into the ration. The basal diet contained (DM basis) 52% forage (with 61% of forage provided as corn silage) and 48% concentrate ingredients. The basal diet was formulated to meet National Research Council (NRC, 2001) requirements for energy and all nutrients except rumen-degradable protein (RDP) and metabolizable protein. Experimental periods lasted 14 d with the first 9 d for adaptation. The basal diet, without urea addition, contained 9.2% RDP in DM and had a predicted RDP balance of −167 g/d (NRC, 2001). There were no effects of dietary treatment on ruminal true digestibility of organic matter or ruminal apparent digestibility of neutral detergent fiber and acid detergent fiber. Total ruminal volatile fatty acid concentrations increased linearly with increasing urea level. Feeding increasing amounts of urea quadratically increased rumen ammonia N concentrations (9.0, 11.9, 12.8, and 17.4 mg/dL at 0, 0.3, 0.6, and 0.9% urea supplementation, respectively), passage of microbial N, and microbial N in duodenal digesta as a percentage of nonammonia N. The results of this study indicate that there were some positive effects of adding urea to the described lactating dairy cow diet, and that microbial protein synthesis was maximized at an average ruminal ammonia N concentration of 12.8 mg/dL when urea was added at 0.6% in diet DM.     

Effect of source of rumen-degraded protein on production and ruminal metabolism in lactating dairy cows

Twenty-eight (8 with ruminal cannulas) lactating Holstein cows were assigned to seven 4 × 4 Latin squares in a 16-wk trial to study the effects on production and ruminal metabolism of feeding differing proportions of rumen-degraded protein (RDP) from soybean meal and urea. Diets contained [dry matter (DM) basis] 40% corn silage, 15% alfalfa silage, 28 to 30% high-moisture corn, plus varying levels of ground dry shelled corn, solvent- and lignosulfonate-treated soybean meal, and urea. Proportions of the soybean meals, urea, and dry corn were adjusted such that all diets contained 16.1% crude protein and 10.5% RDP, with urea providing 0, 1.2, 2.4, and 3.7% RDP (DM basis). As urea supplied greater proportions of RDP, there were linear decreases in DM intake, yield of milk, 3.5% fat-corrected milk, fat, protein, and solids-not-fat, and of weight gain. Milk contents of fat, protein, and solids-not-fat were not affected by source of RDP. Replacing soybean meal RDP with urea RDP resulted in several linear responses: increased excretion of urinary urea-N and concentration of milk urea-N, blood urea-N, and ruminal ammonia-N and decreased excretion of fecal N; there was also a trend for increased excretion of total urinary N. A linear increase in neutral detergent fiber (NDF) digestibility, probably due to digestion of NDF-N from lignosulfonate-treated soybean meal, was observed with greater urea intake. Omasal sampling revealed small but significant effects of N source on measured RDP supply, which averaged 11.0% (DM basis) across diets. Increasing the proportion of RDP from urea resulted in linear decrease in omasal flow of dietary nonammonia N (NAN) and microbial NAN and in microbial growth efficiency (microbial NAN/unit of organic matter truly digested in the rumen). These changes were paralleled by large linear reductions in omasal flows of essential, nonessential, and total amino acids. Overall, these results indicated that replacing soybean meal RDP with that from urea reduced yield of milk and milk components, largely because of depressed microbial protein formation in the rumen and that RDP from nonprotein-N sources was not as effective as RDP provided by true protein.     

Relationships between dry matter content,ensiling, ammonia-nitrogen,and ruminal in vitro starch digestibility in high-moisture corn samples

The objectives of the study were (1) to determine relationships between high-moisture corn (HMC) dry matter (DM), ammonia-N [% of crude protein (CP)], and soluble CP concentrations, and pH, with 7-h ruminal in vitro starch digestibility (ivStarchD), and (2) to evaluate the effect of ensiling on pH, ammonia-N, soluble CP, and ivStarchD measurements in HMC. A data set comprising 6,131 HMC samples (55 to 80% DM) obtained from a commercial feed analysis laboratory was used for this study. Month of sample submittal was assumed to be associated with length of the ensiling period. Data for month of sample submittal were analyzed using Proc Mixed in SAS (SAS Institute Inc., Cary, NC) with month as a fixed effect. Regressions to determine linear and quadratic relationships between ivStarchD and ammonia-N, soluble CP, pH, and DM content were performed using Proc Mixed. The ivStarchD increased by 9 percentage units from October to August of the following year. Similar results were observed for ammonia-N and soluble CP with increases from 1.8 to 4.6% of CP and 31.3 to 46.4% of CP, respectively, from October to August of the following year. Ammonia-N was positively related to ivStarchD (R² = 0.61). The DM content of HMC at silo removal was negatively related (R² = 0.47) to ivStarchD with a decrease of 1.6 percentage units in ivStarchD per 1-percentage-unit increase in DM content. The pH of HMC was negatively related to ammonia-N (R² = 0.53), soluble CP (R² = 0.57), and ivStarchD (R² = 0.51). Combined, ammonia-N, DM, soluble CP, and pH provided a good prediction of ivStarchD (adjusted R² = 0.70). Increasing pH, ammonia-N, soluble CP, and ivStarchD values indicate that HMC may need up to 10 mo of ensiling to reach maximum starch digestibility. Ammonia-N, DM content, soluble CP concentration, and pH are good indicators of ruminal in vitro starch digestibility for high-moisture corn.     

Carbohydrate source and protein degradability alter lactation, ruminal, and blood measures

Thirty-eight lactating dairy cows including 6 ruminally cannulated cows were used in a feeding study to assess effects of feed sources that differed in dietary nonfiber carbohydrate (NFC) composition and ruminal degradability of dietary protein (RDP) on production, ruminal, and plasma measures. The design was a partially balanced, incomplete Latin square with three 21-d periods and a 3 × 2 factorial arrangement of treatments. Samples and data were collected in the last 7 d of each period. Feed sources that differed in NFC profile were dry ground corn (GC; starch), dried citrus pulp (DCP; sugar and pectins), and sucrose+molasses (SM; sugar). Dietary RDP was altered by providing CP with soybean meal (+RDP) or substituting a heat-treated expeller soybean product for a portion of the soybean meal (−RDP). Diets were formulated to be isonitrogenous and similar in NFC concentration. Cows consuming GC had the greatest milk urea nitrogen and milk protein percentage and yield, tended to have the greatest dry matter intake, but had a lesser milk fat percentage compared with cows consuming DCP and SM. Sucrose+molasses diets supported greater dry matter intake, milk protein yield, and 3.5% fat- and protein-corrected milk yield than did DCP diets. On −RDP diets, milk protein percentage was less and milk urea nitrogen and protein yield tended to be less than for +RDP diets. Dry ground corn diverged from DCP and SM in the effect of NFC × RDP, with cows consuming GC having lesser milk yield, 3.5% fat- and protein-corrected milk yield, and efficiency with −RDP as compared with +RDP, whereas these production measures were greater with −RDP than +RDP for cows consuming DCP and SM. In contrast, in situ NDF digestibility at 30 h for GC and SM was greater for −RDP as compared with +RDP, but the reverse was true for DCP. The lowest ruminal pH detected by 6 h postfeeding was also influenced by the interaction of NFC × RDP, with cows consuming SM having a lower pH with +RDP than with −RDP and cows consuming DCP having a similar pH on either RDP treatment. Total rumen volatile fatty acid concentrations did not differ among diets, but acetate molar percent was greater for DCP than for SM, and GC had the lowest molar percent for butyrate and valerate and greatest branched-chain volatile fatty acid concentration. Valerate molar percent and NH₃ concentration tended to be greater with +RDP than with −RDP. Plasma glucose and insulin were both greater in cows receiving SM than in those receiving DCP. Protein degradability, NFC source, and their interactions affected lactation, ruminal, and blood measures, suggesting that these dietary factors warrant further consideration in diet formulation.     

Technical note: Evaluation of markers for estimating duodenal digesta flow and ruminal digestibility: Acid detergent fiber,sulfuric acid detergent lignin,and n-alkanes

The amount of digesta flowing to the duodenum is a relevant measurement for the evaluation of nutrient supply to ruminants, which is usually estimated in animals fitted with a duodenal T-type cannula using internal or external markers. This study evaluated acid detergent fiber (ADF) compared with external (C₃₂n-alkane) and internal [sulfuric acid lignin (ADL) and n-alkanes C₃₁ and C₃₃] markers for estimating duodenal flow and(or) ruminal digestibility of dry matter (DM) in cattle and sheep. In the first assay, 4 duodenally cannulated Holstein steers housed in metabolism cages, dosed with C₃₂n-alkane, and fed Avena strigosa plus concentrate and increasing levels of tannin extract to reduce ruminal digestibility, were used in a Latin square design. The mobile-bag technique was used to measure the intestinal disappearance of ADL and ADF from forage (Avena strigosa, Pennisetum purpureum, Cynodon dactylon, and Medicago sativa) and concentrate (corn grain, soybean meal, and sunflower meal) samples that were previously incubated in the rumen of additional fistulated steer for 12, 24, 36, or 48 h. The ADF concentration in residues recovered in the feces was strongly related to the ADF concentration in residues at the duodenum (R² = 0.93, standard deviation = 30.0, n = 901). This relationship showed a lower precision for ADL fraction (R² = 0.88, standard deviation = 12.6, n = 590). In a second assay, duodenal flow and ruminal DM digestibility were calculated from the duodenal and fecal concentration of either marker. We observed a significant effect of marker type on ruminal DM digestibility values, and the effect of tannin treatments was observed only when ADF or ADL was used as the marker. The lowest residual error was obtained for ADF. Ruminal DM digestibility was, on average, higher for C₃₁ and C₃₃n-alkanes, and the use of dosed C₃₂n-alkane resulted in a negative value. In the third assay, a data set of 235 individual observations was compiled from digestibility trials to compare ADF and ADL as markers for estimating duodenal digesta flow in wethers (n = 204) and cattle (n = 31). We observed a strong relationship between markers (R² = 0.84 in sheep and R² = 0.88 in cattle), but variance analysis within trials indicated that ADF was more precise than ADL. In conclusion, in digestibility trials in which fecal output was measured and spot samples of the duodenal digesta were obtained, duodenal flow and ruminal digestibility of the DM may be estimated from the relationship between the ADF concentration in feces and that in the duodenal digesta of ruminants.     

Effect of dietary crude protein concentration on milk production and nitrogen utilization in lactating dairy cows

Forty lactating Holstein cows, including 10 with ruminal cannulas, were blocked by days in milk into 8 groups and then randomly assigned to 1 of 8 incomplete 5 × 5 Latin squares to assess the effects of 5 levels of dietary crude protein (CP) on milk production and N use. Diets contained 25% alfalfa silage, 25% corn silage, and 50% concentrate, on a dry matter (DM) basis. Rolled high-moisture shelled corn was replaced with solvent-extracted soybean meal to increase CP from 13.5 to 15.0, 16.5, 17.9, and 19.4% of DM. Each of the 4 experimental periods lasted 28 d, with 14 d for adaptation and 14 d for data collection. Spot sampling of ruminal digesta, blood, urine, and feces was conducted on d 21 of each period. Intake of DM was not affected by diet but milk fat content as well as ruminal acetate, NH₃, and branched-chain volatile fatty acids, urinary allantoin, and blood and milk urea all increased linearly with increasing CP. Milk and protein yield showed trends for quadratic responses to dietary CP and were, respectively, 38.3 and 1.18 kg/d at 16.5% CP. As a proportion of N intake, urinary N excretion increased from 23.8 to 36.2%, whereas N secreted in milk decreased from 36.5 to 25.4%, as dietary protein increased from 13.5 to 19.4%. Under the conditions of this study, yield of milk and protein were not increased by feeding more than 16.5% CP. The linear increase in urinary N excretion resulted from a sharp decline in N efficiency as dietary CP content increased.     

Quantifying ruminal nitrogen metabolism using the omasal sampling technique in cattle—A meta-analysis

Mixed model analysis of data from 32 studies (122 diets) was used to evaluate the precision and accuracy of the omasal sampling technique for quantifying ruminal-N metabolism and to assess the relationships between nonammonia-N flow at the omasal canal and milk protein yield. Data were derived from experiments in cattle fed North American diets (n = 36) based on alfalfa silage, corn silage, and corn grain and Northern European diets (n = 86) composed of grass silage and barley-based concentrates. In all studies, digesta flow was quantified using a triple-marker approach. Linear regressions were used to predict microbial-N flow to the omasum from intake of dry matter (DM), organic matter (OM), or total digestible nutrients. Efficiency of microbial-N synthesis increased with DM intake and there were trends for increased efficiency with elevated dietary concentrations of crude protein (CP) and rumen-degraded protein (RDP) but these effects were small. Regression of omasal rumen-undegraded protein (RUP) flow on CP intake indicated that an average 32% of dietary CP escaped and 68% was degraded in the rumen. The slope from regression of observed omasal flows of RUP on flows predicted by the National Research Council (2001) model indicated that NRC predicted greater RUP supply. Measured microbial-N flow was, on average, 26% greater than that predicted by the NRC model. Zero ruminal N-balance (omasal CP flow = CP intake) was obtained at dietary CP and RDP concentrations of 147 and 106 g/kg of DM, corresponding to ruminal ammonia-N and milk urea N concentrations of 7.1 and 8.3 mg/100 mL, respectively. Milk protein yield was positively related to the efficiency of microbial-N synthesis and measured RUP concentration. Improved efficiency of microbial-N synthesis and reduced ruminal CP degradability were positively associated with efficiency of capture of dietary N as milk N. In conclusion, the results of this study indicate that the omasal sampling technique yields valuable estimates of RDP, RUP, and ruminal microbial protein supply in cattle.     

Effect of dietary crude protein concentration on ruminal nitrogen metabolism in lactating dairy cows

Ten lactating Holstein cows fitted with ruminal cannulas that were part of a larger feeding trial were blocked by days in milk into 2 groups and then randomly assigned to 1 of 2 incomplete 5 × 5 Latin squares. Diets contained [dry matter (DM) basis] 25% alfalfa silage, 25% corn silage, and 50% concentrate. Rolled high-moisture shelled corn was replaced with solvent-extracted soybean meal to increase crude protein (CP) from 13.5% to 15.0, 16.5, 17.9, and 19.4% of DM. Each of the 4 experimental periods lasted 28 d with data and sample collection performed during the last 8 d. Digesta samples were collected from the omasum to quantify the ruminal outflow of different N fractions. Intake of DM was not affected but showed a quadratic trend with maxima of 23.9 kg/d at 16.5% CP. Ruminal outflow of total bacterial nonammonia N (NAN) was not different among diets but a significant linear effect of dietary CP was detected for this variable. Bacterial efficiency (g of total bacterial NAN flow/kg of organic matter truly digested in the rumen) and omasal flows of dietary NAN and total NAN also showed positive linear responses to dietary CP. Total NAN flow increased from 574 g/d at 13.5% CP to 688 g/d at 16.5% CP but did not increase further with the feeding of more CP. Under the conditions of this study, 16.5% of dietary CP appeared to be sufficient for maximal ruminal outflow of total bacterial NAN and total NAN.     

Effects of spontaneous heating on forage protein fractions and in situ disappearance kinetics of crude protein for alfalfa-orchardgrass hays packaged in large round bales

During 2006 and 2007, forages from 3 individual hay harvests were used to assess the effects of spontaneous heating on concentrations of crude protein (CP), neutral detergent insoluble CP (NDICP), acid detergent insoluble CP (ADICP), and in situ disappearance kinetics of CP and NDICP for large round bales of mixed alfalfa (Medicago sativa L.) and orchardgrass (Dactylis glomerata L.). Over the 3 harvests, 96 large round bales were made at preset bale diameters of 0.9, 1.2, or 1.5 m and at moisture concentrations ranging from 9.3 to 46.6%. Internal bale temperatures were monitored daily during an outdoor storage period. The change in concentrations of NDICP (poststorage − prestorage) increased with heating degree days (HDD) >30°C in a relationship best explained with a nonlinear model {Y = 24.9 - [22.7 × (e^{−0.000010 × x × x})]; R² = 0.892} that became asymptotic at +24.9 percentage units of CP, thereby indicating that NDICP increases rapidly within bales that heat spontaneously. When maximum internal bale temperature (MAX) was used as the independent variable, the best regression model was quadratic and the coefficient of determination was still relatively high (R² = 0.716). The change in concentrations of ADICP (poststorage − prestorage; ΔADICP) also increased with HDD and was best fitted to a nonlinear model {Y = 14.9 - [15.7 × (e^{−0.0000019 × x × x})]} with a very high coefficient of determination (R² = 0.934). A similar quartic response was observed for the regression of ΔADICP on MAX (R² = 0.975). Increases in ΔADICP as a result of heating (HDD or MAX) were paralleled by concurrent increases in hemicellulose at relatively low increments of heating, but the inverse relationship was observed as hemicelluloses likely became reactive and concentrations decreased in more severely heated hays. Changes in ruminal disappearance rate of CP were best fitted to cubic models for regressions on both HDD (R² = 0.939) and MAX (R² = 0.876); these changes represented an approximate 50% rate reduction in severely heated hays relative to prestorage controls. Within ranges of heating most commonly encountered under field conditions, changes in rumen-degradable protein decreased in a primarily linear relationship with HDD or MAX. However, the mean change in rumen-degradable protein for the 4 most severely heated hays was only −2.6 percentage units of CP, which represents a minimal reduction from prestorage controls and is far less than the maximum of −7.9 percentage units of CP observed with less-severe heating. Interpretation of these results was complicated by poor recovery of NDICP from our most severely heated hays following machine rinsing of 0-h Dacron bags; theoretically, and by definition, this unrecovered pool of NDICP is assumed to be entirely degradable in the rumen. It remains unclear whether these responses could be corroborated in vivo or by other analytical techniques, or whether the magnitude of HDD or MAX for our most severely heated hays exceeds the reliable limits for estimating RDP via in situ methodology.     

Subacute ruminal acidosis challenge changed in situ degradability of feedstuffs in dairy goats

This study investigated the effects of wheat-induced subacute ruminal acidosis (SARA) on rumen bacterial populations and in situ degradabilities of NDF, starch, and crude protein of feeds. Four multiparous dairy goats (BW = 60 ± 3.3 kg) fitted with ruminal cannulas were assigned to a 2 × 2 crossover design (28-d treatment periods separated by a 7-d washout interval). The treatment diets consisted of 2 levels of cracked wheat: 0 (control, corn based concentrate) and 35% (diet-induced SARA, wheat-based concentrate), with a constant forage- (45% alfalfa hay and 5% corn silage of DM) to-concentrate (50% of DM) ratio. Results indicate that diets with a 35% wheat decreased ruminal pH (6.21 vs. 5.98) and increased the duration (1.13 vs. 4.72 h/d) and area (0.12 vs. 0.78 pH × h/d) of ruminal pH below 5.6 and induced SARA. The SARA increased ruminal total volatile fatty acid concentration, from 105.0 to 123.8 mM, and decreased the acetate molar proportion (62.8 vs. 56.6 mol/100 mol) and the acetate-to-propionate ratio (3.5 vs. 2.8). Compared with the control group, SARA decreases the relative abundance of Fibrobacter succinogenes (−59.3%) and Ruminococcus flavefaciens (−68.4%), whereas it increased Succinimonas amylolytica (198.1%) and Ruminobacter amylophilus (125.2%). The SARA decreased 24- and 48-h dry matter (DM) and neutral detergent fiber (NDF) degradabilities of corn silage. The 48-h degradabilities of DM (51.0 vs. 48.2%) and NDF (40.3 vs. 36.0%) in alfalfa hay were not affected by SARA, but the SARA tended to reduce the 24-h DM (49.6 vs. 46.3%) and NDF (37.8 vs. 33.2%) degradabilities. The effective ruminal degradabilities of DM and NDF in alfalfa hay and corn silage were reduced during SARA. In situ degradability parameters of DM and starch of wheat were not affected by SARA, but starch degradability of corn (9.5 vs. 13.3%/h) increased. The SARA reduced in situ 12-h degradabilities of DM and crude protein of soybean meal and extruded soybean without affecting the degradabilities of the other protein supplements (corn gluten meal, cottonseed meal, corn dried distillers grains with solubles, rapeseed meal, and wheat germ meal). These results indicated that the cracked wheat-induced SARA reduced the degradation of NDF in roughages and that of protein in soybean meal (−19.8%) and extruded soy (−18.9%) and increased the starch degradability in corn, due to the increased amylolytic bacteria and decreased cellulolytic bacteria counts in the rumen.     

Effect of dietary level of rumen-degraded protein on production and nitrogen metabolism in lactating dairy cows

Twenty-eight (8 with ruminal cannulas) lactating Holstein cows were assigned to 4 × 4 Latin squares and fed diets with different levels of rumen-degraded protein (RDP) to study the effect of RDP on production and N metabolism. Diets contained [dry matter (DM) basis] 37% corn silage, 13% alfalfa silage, and 50% concentrate. The concentrate contained solvent and lignosulfonate-treated soybean meal and urea, and was adjusted to provide RDP at: 13.2, 12.3, 11.7, and 10.6% of DM in diets A to D, respectively. Intake of DM and yield of milk, fat-corrected milk, and fat were not affected by treatments. Dietary RDP had positive linear effects on milk true protein content and microbial non-ammonia N (NAN) flow at the omasal canal, and a quadratic effect on true protein yield, with maximal protein production at 12.3% RDP. However, dietary RDP had a positive linear effect on total N excretion, with urinary N accounting for most of the increase, and a negative linear effect on environmental N efficiency (kg of milk produced per kg of N excreted). Therefore, a compromise between profitability and environmental quality was achieved at a dietary RDP level of 11.7% of DM. Observed microbial NAN flow and RDP supply were higher and RUP flow was lower than those predicted by the NRC (2001) model. The NRC (2001) model overpredicted production responses to RUP compared with the results in this study. Replacing default NRC degradation rates for protein supplements with rates measured in vivo resulted in similar observed and predicted values, suggesting that in situ degradation rates used by the NRC are slower than apparent rates in this study.     

Milk from forage as affected by rumen degradable protein and corn grinding when feeding corn- and alfalfa silage-based diets

To increase the production of milk from forage (MF), a previous experiment with alfalfa silage showed the importance of a complementary combination of concentrates and forages offered. When corn silage is fed with alfalfa, increasing the rumen degradable protein (RDP) content in the diet should allow a better utilization of forage energy. To evaluate this hypothesis, 8 multiparous Holstein cows in early lactation were used in a replicated 4 × 4 Latin square design with 3-wk periods. Diets were fed as total mixed rations and were formulated to provide similar levels of net energy for lactation and crude protein but differing in RDP. Corn and alfalfa silages were used. Treatments were: 1) cracked corn-based concentrate providing low RDP [level recommended by the NRC (2001); RDP = 11.1% of dry matter (DM)]; 2) cracked corn-based concentrate providing medium RDP (RDP = 12.8% of DM); 3) cracked corn-based concentrate providing high RDP (RDP = 14.5% of DM); and 4) ground corn-based concentrate providing high RDP (RDP = 13.6% of DM). The first 3 treatments, using cracked corn, were compared on the basis of their RDP level. For these treatments, MF, calculated on a protein basis, decreased and the average of MF calculated on an energy basis and MF calculated on a protein basis tended to decrease as RDP increased. There was no difference for MF calculated on an energy basis between treatments. Increasing dietary RDP levels decreased the milk yield (from 32.8 to 30.7 kg/d) and milk protein yield (from 1,094 to 1,005 g/d) but not the milk fat yield. The milk urea N concentration increased as RDP increased. This suggests that there is no advantage of feeding RDP above the NRC recommendations when diets are based on corn and alfalfa silage. At high RDP levels (treatments 3 and 4), ground corn supported higher DM intake and yields of milk and protein than did cracked corn. Milk from forage, calculated on a protein basis, was higher and milk urea N decreased with ground corn. Even with corn silage in the diet, grinding corn grain proved to be beneficial to milk yield and MF production.     

Substitution of molasses for corn grain at two levels of degradable protein. II. Effects on ruminal fermentation,digestion, and nitrogen metabolism

Our objective was to evaluate cow N metabolism and ruminal measures with diets containing 3 different levels of molasses or finely ground dry corn grain with 2 levels of ruminally degradable protein (RDP). Twelve lactating, ruminally cannulated Holstein cows (parity 2.25 ± 0.62; 185 ± 56 DIM; 41.3 ± 6.3 kg of milk initially) were individually fed in an experiment designed as a split-plot, replicated 3 × 3 Latin square, where each period lasted 28 d. Six diets were formulated according to a 2 × 3 factorial arrangement of treatments, where 2 levels of RDP (+RDP and ?RDP) were fed throughout the experiment as the whole plot, and 3 levels of molasses (0, 5.25, or 10.50% of dry matter replacing finely ground dry corn grain) were fed in sequences of the Latin squares. Dry matter intake did not differ by diet, although ash intake increased linearly with increasing molasses. Ruminal pH, organic acid concentration, and ammonia concentration were not affected by diet. Molar percentages of ruminal acetate decreased and butyrate increased linearly with increasing levels of molasses. Ruminal free amino acid concentration was greater for +RDP, whereas branched-chain volatile fatty acids declined linearly with increasing molasses. Rumen content mass, ruminal liquid, and ruminal acetate pool size was greater for ?RDP, although ruminal lactate pool size tended to be greater with +RDP. Increased ruminal lactate when increasing molasses with high RDP should be explored further, to optimize microbial efficiency and rumen health. Total-tract apparent dry matter digestibility based on spot sampling was not affected by diet; however, ash digestibility increased linearly with increasing levels of molasses. Calculated urine output was greater for cows fed diets with increasing levels of molasses and for cows fed +RDP. Grams of N distributed to excretion pools were not different across diets, although, as molasses increased, a lower proportion of N intake was excreted in urine. Overall, the results from this experiment showed that dairy cows used dietary carbohydrates differently during ruminal fermentation, with increasing molasses resulting in increased butyrate molar proportions at the expense of acetate. Additionally, RDP tended to modify the effects of carbohydrate fermentation, resulting in a tendency for increasing lactate molar pool size only in diets with greater RDP, although this did not ultimately affect ruminal pH.     

Effects of physically effective fiber on digestive processes and milk fat content in early lactating dairy cows fed total mixed rations

Data from recent research studies were analyzed quantitatively, and the random effect of experiment was assessed to define the physiological responses of dairy cows in early lactation to intake of physically effective neutral detergent fiber (peNDF). All studies were conducted with lactating Holstein cows (84.8 ± 3.54 days in milk) in Latin square designs, and feeds were offered ad libitum as total mixed rations (TMR). The peNDF was estimated by 2 measurement techniques, the NDF content of TMR multiplied by amount of dry matter (DM) retained on a 1.18-mm screen (peNDF_{> 1.18}) and NDF content of TMR multiplied by the proportion of DM retained by 19- and 8-mm Penn State Particle Separator screens (peNDF_{> 8}). Other factors, including concentrations of NDF, forage NDF, non-fiber carbohydrates, the amount of digestible organic matter of forages (FDOM), and the intake of ruminally degradable starch (RDSI) from grain in the diet were also investigated. The studied animal response variables included feed intake, ruminal fermentation, chewing activity, fiber digestibility, and milk production and composition. The ruminal pH (day mean) in this study ranged from 5.30 to 6.59. Using peNDF_{> 1.18} approach, the requirements for physically effective fiber in high-yielding dairy cows fed TMR in an ad libitum intake were estimated to be about 19% of ration DM or 4.1 kg/d or 0.6 kg/100 kg of body weight to maintain a ruminal pH of about 6.0. When peNDF was measured as peNDF_{> 8}, ruminal pH responded in a quadratic fashion but the confidence of estimation was lower (R² = 0.27) compared with the peNDF_{> 1.18} approach (R² = 0.67). Results of these data analyses showed that peNDF_{> 1.18} provided a satisfactory estimation of the mean ruminal pH (R² = 0.67) and NDF digestibility (R² = 0.56). Furthermore, peNDF_{> 1.18} was poorly, although positively, correlated to daily chewing (R² = 0.17), and rumination (R² = 0.24) activity. On the other hand, results from these analyses showed that milk parameters are less sensitive to the effects of dietary peNDF than other variables, such as ruminal pH, chewing activity, and fiber digestibility. Dietary FDOM correlated positively (moderately) to ruminal pH (R² = 0.24), daily chewing (R² = 0.23), and rumination (R² = 0.29) activity, whereas the daily RDSI from grain correlated negatively to ruminal pH (R² = 0.55) and positively to total volatile fatty acids (R² = 0.27). Inclusion of FDOM and RDSI from grain along with peNDF_{> 1.18} in the models that predict rumen pH further improved the accuracy of prediction. This approach appeared to further complement the concept of peNDF that does not account for differences in ruminal fermentability of feeds.     

Effect of amount and ruminal degradability of soybean meal protein on performance of lactating dairy cows

Twenty-eight Holstein cows (4 with ruminal cannulas) were blocked by days in milk into 7 groups and then randomly assigned to 1 of 7 balanced 4 × 4 Latin square diet sequences. The diets contained [dry matter (DM) basis] 20% alfalfa silage, 35% corn silage, and 45% concentrate mainly from high-moisture corn and soybean meal. Diets differed in crude protein (CP) content and source of protein supplement: diet A) 15.6% CP, 3.7% solvent-extracted soybean meal (SSBM), 4.5% expeller soybean meal (ESBM); diet B) 16.6% CP, 9.6% SSBM, 0% ESBM; diet C) 16.6% CP, 4.6% SSBM, 5.9% ESBM; and diet D) 17.6% CP, 11.7% SSBM, 0% ESBM. Each experimental period consisted of 14 d for adaptation plus 14 d for collection of production data. Sampling of ruminal digesta and spot sampling of blood, feces, and urine was done on d 26 and 27 of each period. Planned contrasts compared included diet A vs. diet B, diet B vs. diet C, and diet B vs. diet D. There were no effects of diet on most of the production traits measured. However, milk yield tended to be higher for diet B vs. A. Trends were also detected for higher DM intake and weight gain and lower milk yield/DM intake in cows fed diet D vs. diet B. Milk lactose content was higher on diets A and C than on B. Ruminal NH₃ was higher on diet D vs. B, but other ruminal metabolites, apparent nutrient digestibility, and estimated bacterial CP synthesis did not differ across diets. Blood and milk urea-N were higher on diets C and D than on B; milk urea-N was higher on diet B than on A. Increasing dietary CP from 16.6% (diet B) to 17.6% (diet D) increased urinary N excretion by 54 g/d and reduced apparent N efficiency (milk N/N intake) by 2.5 percentage units, without altering yield. Under the conditions of this trial, milk production was not improved by feeding rumen-undegraded protein from ESBM or greater amounts of rumen-degraded protein from SSBM. Feeding more than 16.6% CP depressed N efficiency.     

Ruminal degradability of dry matter,crude protein,and amino acids in soybean meal,canola meal,corn, and wheat dried distillers grains

Different protein sources, such as canola meal (CM) or dried distillers grains (DDG), are currently used in dairy rations to replace soybean meal (SBM). However, little data exists comparing their rumen degradation in a single study. Therefore, the objective of this study was to compare the ruminal degradation of dry matter (DM), crude protein (CP), and AA of SBM, CM, high-protein corn DDG (HPDDG), and wheat DDG plus solubles (WDDGS). In situ studies were conducted with 4 rumen-fistulated lactating Holstein cows fed a diet containing 38% grass hay and 62% corn-based concentrate. Each protein source was incubated in the rumen of each cow in nylon bags for 0, 2, 4, 8, 16, 24, and 48 h to determine DM and CP rumen degradation kinetics, whereas additional bags were also incubated for 16 h to evaluate AA ruminal disappearance. Rumen DM and CP degradability was calculated from rumen-undegraded residues corrected or not for small particle loss. Data were fitted to an exponential model to estimate degradation parameters and effective degradability (ED) was calculated with a passage rate of 0.074 h⁻¹. The WDDGS and SBM had higher uncorrected ED (DM = 75.0 and 72.6%; CP = 84.8 and 66.0%, respectively) than CM and HPDDG (DM = 57.2 and 55.5%; CP = 59.3 and 48.2%, respectively), due to higher soluble fraction in WDDGS and a combination of higher potentially degradable fraction and rate of degradation in SBM. Correction for small particle loss from bags, higher for WDDGS than for the other protein sources, decreased estimated ED but did not alter feed ranking. The ruminal disappearance of AA after 16 h of incubation reflected the overall pattern of CP degradation between protein supplements, but the ruminal disappearance of individual AA differed between protein supplements. Overall, these results indicate that, in the current study, (1) SBM and WDDGS were more degradable in the rumen than CM and HPDDG, and (2) that small particle loss correction is relevant but does not alter this ranking.     

Effects of time at suboptimal pH on rumen fermentation in a dual-flow continuous culture system

Ruminal pH varies considerably during the day, achieving values below 6.0 when cows consume large amounts of concentrates. Low ruminal pH has negative effects on ruminal fermentation. However, previous studies have indicated that rumen bacteria may resist short periods of low ruminal pH, and it is not clear how long this period may be before rumen microbial fermentation is negatively affected. Seven dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods with the same diet (97 g of dry matter/d of a 60:40 forage-to-concentrate diet, 18.3% crude protein, 35.9% neutral detergent fiber), temperature (39°C), and solid (5%/h) and liquid (10%/h) dilution rates to study the effects of increasing time at suboptimal pH on rumen microbial fermentation and nutrient flow. Treatments were a constant pH of 6.4 and 6 different intervals of time during the day (4, 8, 12, 16, 20, 24 h) at suboptimal pH (5.5), with the rest of the day being at pH 6.4. Polynomial equations were derived using the Mixed procedure of SAS, and linear, quadratic and cubic terms were left in the equation if P < 0.10. True organic matter digestion decreased with increasing time at suboptimal pH and was best described by a cubic regression (TOMD = 58.5 − 2.15x + 0.16x² −0.0037x³; R² = 0.74). Digestion of NDF (DNDF = 55.1 − 1.00x; R² = 0.75) and digestion of ADF (DADF = 56.2 − 1.33x; R² = 0.78) decreased linearly with increasing time at suboptimal pH. Total VFA had a cubic response (VFA = 112.7 − 2.09x + 0.17x² − 0.0054x³; R² = 0.82). The proportion of acetate decreased linearly (acetate = 58.7 − 0.61x; R² = 0.79). The propionate proportion increased (propionate = 17.6 + 2.09 × −0.044x²; R² = 0.85) and branched-chain VFA decreased (BCVFA = 4.45 −0.51x + 0.014x²; R² = 0.75) quadratically. The ammonia N concentration (NH₃-N = 5.85 − 0.13x; R² = 0.46) and flow (NH₃-N flow = 0.18 − 0.0039x; R² = 0.43) decreased linearly as the time at suboptimal pH increased. Crude protein degradation (CPd = 41.9 − 1.60x + 0.060x²; R² = 0.71), efficiency of microbial protein synthesis (EMPS = 26.6 − 0.33x + 0.021x²; R² = 0.77), microbial N flow (MN flow = 1.38 − 0.036x + 0.0015x²; R² = 0.77), and dietary N flow (DN flow = 1.49 + 0.041x − 0.0015x²; R² = 0.65) had a quadratic response. The flow of essential, nonessential, and most individual AA increased linearly with increasing time at suboptimal pH. The effects of pH on rumen fermentation appear to start as soon as pH drops to suboptimal pH.     

Comparison of four markers for quantifying microbial protein flow from the rumen of lactating dairy cows

Eight ruminally cannulated lactating cows from a study on the effects of dietary rumen degraded protein (RDP) on production and N metabolism were used to compare ¹⁵N, total purines, amino acid (AA) profiles, and urinary excretion of purine derivatives (PD) as microbial markers for quantifying the flow of microbial protein at the omasal canal. Dietary RDP was gradually decreased by replacing solvent soybean meal and urea with lignosulfonate-treated soybean meal. The purine metabolites xanthine and hypoxanthine were present in digesta and microbial samples and were assumed to be of microbial origin. The sum of the purines and their metabolites (adenine, guanine, xanthine, and hypoxanthine) were defined as total purines (TP) and used as a microbial marker. Decreasing dietary RDP from 13.2 to 10.6% of dry matter (DM) reduced microbial nonammonia N (NAN) flows estimated using TP (from 415 to 369 g/d), ¹⁵N (from 470 to 384 g/d), AA profiles (from 392 to 311 g/d), and PD (from 436 to 271 g/d). Averaged across diets, microbial NAN flows were highest when estimated using TP and ¹⁵N (398 and 429 g/d), lowest when using PD (305 g/d), and intermediate when using AA profiles (360 g/d) as microbial markers. Correlation coefficients between ¹⁵N and TP for fluid-associated bacteria, particle-associated bacteria, and total microbial NAN flows were 0.38, 0.85, and 0.69, respectively. When TP was used as the microbial marker, ruminal escape of dietary NAN was not affected by replacing solvent soybean meal with lignosulfonate-treated soybean meal in the diets. The direction and extent of response of dietary and microbial NAN flow to dietary treatments were similar when estimated using ¹⁵N, AA profiles, and PD, and were in agreement with previously published data and National Research Council predictions. Microbial and dietary NAN flows from the rumen estimated using ¹⁵N appeared to be more accurate and precise than the other markers. Caution is required when interpreting results obtained using TP as the microbial marker.     

Milk production of dairy cows fed differing concentrations of rumen-degraded protein

Thirty-two multiparous and 16 primiparous Holstein cows in midlactation averaging 126 d in milk were used to determine the effects of rumen-degraded protein (RDP) concentration on lactation performance. Cows were assigned to diets in a repeated Latin square design with 3-wk experimental periods. Diets were formulated to provide 4 concentrations of dietary RDP [6.8, 8.2, 9.6, and 11.0% of dry matter (DM)] while rumen-undegraded protein remained constant (5.8% of DM). Diets contained 50% corn silage and 50% concentrate (DM basis). Ingredients within diets were equal across treatments except for ground corn, soybean meal, and ruminally protected soybean meal. Dry matter intake was not affected by treatment. Milk yield, fat yield, and protein yield all increased linearly when cows were fed diets with greater RDP. Milk fat and protein concentration each increased by 0.16 percentage units for cows fed 11% RDP compared with 6.8% RDP. Milk protein yield increased by 0.19 g/d for every 1 g/d increase in crude protein supplied mainly as RDP. As RDP increased, the efficiency of N use declined linearly. Milk urea N increased linearly when cows were fed increasing amounts of RDP, indicating increased losses of N via urine. Feeding deficient RDP diets to dairy cows can decrease nitrogen excretion, but it also decreases lactation performance. These data show an environmental benefit from underfeeding RDP to dairy cows according to National Research Council requirements, but at a financial cost to the dairy producer.     

Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae fermentation product

The rumen microbial ecosystem is a critical factor that links diets to bovine physiology and productivity; however, information about dietary effects on microbial populations has generally been limited to small numbers of samples and qualitative assessment. To assess whether consistent shifts in microbial populations occur in response to common dietary manipulations in dairy cattle, samples of rumen contents were collected from 2 studies for analysis by quantitative real-time PCR (qPCR). In one study, lactating Holstein cows (n = 8) were fed diets in which a nonforage fiber source replaced an increasing proportion of forages and concentrates in a 4 × 4 Latin square design, and samples of ruminal digesta were collected at 9-h intervals over 3 d at the end of each period. In the second study, lactating Holstein cows (n = 15) were fed diets with or without the inclusion of a Saccharomyces cerevisiae fermentation product (SCFP) in a crossover design. In this study, rumen liquid and solid samples were collected during total rumen evacuations before and after feeding in a 42-h period. In total, 146 samples of ruminal digesta were used for microbial DNA isolation and analysis by qPCR. Validated primer sets were used to quantify total bacterial and anaerobic fungal populations as well as 12 well-studied bacterial taxa. The relative abundance of the target populations was similar to those previously reported. No significant treatment effects were observed for any target population. A significant interaction of treatment and dry matter intake was observed, however, for the abundance of Eubacterium ruminantium. Increasing dry matter intake was associated with a quadratic decrease in E. ruminantium populations in control animals but with a quadratic increase in E.ruminantium populations in cows fed SCFP. Analysis of sample time effects revealed that Fibrobacter succinogenes and fungal populations were more abundant postfeeding, whereas Ruminococcus albus tended to be more abundant prefeeding. Seven of the target taxa were more abundant in either the liquid or solid fractions of ruminal digesta. By accounting for the total mass of liquid and solid fractions in the rumen and the relative abundance of total bacteria in each fraction, it was estimated that 92% of total bacteria were found in the solid digesta fraction.