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We hypothesized that dairy cows fed oscillating metabolizable protein (MP) and crude protein (CP) concentrations on a 24-h frequency for a diet formulated to be below MP requirements would use N more efficiently (i.e., increased milk protein yields and less manure N) without increasing mobilization of body protein stores than would cows fed the same deficient MP diet continuously, although both treatments would on average have equal MP concentrations. In a randomized block design, 30 Holstein cows (119 ± 21 d in milk; 667 ± 69 kg of body weight) were blocked according to milk yield within a parity (3 primiparous and 7 multiparous blocks) and fed 1 of 3 treatments: (1) diet with 16.2% CP (109% of MP requirements) fed continuously (109MP), (2) diet with 14.1% CP (95% of MP requirements) fed continuously (95MP), or (3) diets oscillating on a 24-h cycle from the 109MP diet and a diet with 11.9% CP (∼78% of MP requirements) such that average CP and MP concentration would be the same as 95MP (OSC). Dry matter intake was similar between 109MP and 95MP (22.9 vs. 23.2 kg/d) but tended to be lower for OSC (22.2 kg/d) compared with 95MP. Milk yield was greater for 109MP compared with 95MP (36.6 vs. 35.1 kg/d) and similar between 95MP and OSC (35.3 kg/d). Milk protein and energy-corrected milk yields were similar among treatments. Milk urea N (MUN) concentration was higher for 109MP compared with 95MP (12.8 vs. 10.2 mg/dL), and tended to be higher for OSC (10.9 mg/dL) compared with 95MP. Higher MUN concentration for OSC occurred despite lower N intake (474 vs. 512 g of N/d) and similar milk N outputs compared with 95MP (164 vs. 179 g/d). On days when cows on OSC were fed high versus low MP diets, yields of milk (34.8 vs. 36.3 kg/d) and milk protein (1.0 vs. 1.1 kg/d) and MUN concentration (9.3 vs. 12.5 mg/dL) followed the oscillation pattern but lagged the change in diet CP by 1 d, whereas dry matter intake, yields of milk fat, plasma energy metabolites, AA, and 3-methyl-His were similar between days. Nutrient digestibility was similar for major nutrients across treatments except for CP, which was greater for 109MP (65.2%) and OSC (65.3%) compared with 95MP (61.7%). Compared with 95MP, OSC did not increase milk N relative to N intake (averaged 0.35 g of milk N/g of N intake) or N balance; however, urinary N output was increased for OSC versus 95MP (0.32 vs. 0.24 g of urine N/g of N intake). Body composition estimated using urea dilution was similar across treatments, and all cows accreted lipid and energy during the trial. Empty body CP did not change over the 50-d treatment period. Overall, greater CP digestion, urinary N excretion, and MUN concentrations with lesser N intake and similar milk N outputs for OSC compared with 95MP suggests that the lower energy intake by OSC cows may have limited potential responses to altered N metabolism., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Nitrogen concentrations in feeds, feces, milk, and urine samples were measured using 2 analytical methods following different drying procedures. Ten samples of corn silage, alfalfa silage, and concentrates collected from 2017 to 2018 at Krauss Dairy Research Center, The Ohio State University (Wooster), were used. A 4-d total collection digestion trial provided fecal samples from 10 cows (1 sample/cow), and another 10 cows were used to collect milk samples (1 sample/cow) and spot urine samples (1 sample/cow). Spot urine samples were acidified immediately to pH <3.0 when collected. Feed samples were oven dried (55°C) or lyophilized and analyzed using the Kjeldahl (KJ; copper sulfate as a catalyst) method and a combustion method (elemental analyzer; EA). Feces, urine, and milk samples were analyzed for N using the following methods: (1) fresh samples by KJ (referred to as wet KJ), (2) lyophilization (urine and milk for 8 h; feces for 120 h) followed by EA (LYO-EA), and (3) oven drying (milk and urine for 1 h; feces for 72 h at 55°C) followed by EA (OD-EA). Additionally, changes in N content of acidified urine at -20° over 180 d of storage were examined. Nitrogen concentrations in corn silage, alfalfa silage, and concentrates were greater for EA by 6.1, 4.8, and 8.3%, respectively, compared with KJ. Analysis of dried samples via EA compared with wet KJ resulted in lower fecal N content (27.8 vs. 29.3 g/kg of DM). Nitrogen concentration in fecal samples via KJ after lyophilization was lower by 5% compared with wet KJ but did not differ from LYO-EA, suggesting that N losses occurred during drying. Nitrogen determination with EA after drying of samples resulted in greater milk N (5.70 vs. 5.50 g/kg) and urinary N (9.16 vs. 9.06 g/kg) content compared with wet KJ. However, drying method (i.e., lyophilization vs. oven drying) did not affect N content of milk, urine, or feces. The use of EA resulted in lower percentage deviation of N content from duplicate sample assays for most samples (no difference was found for concentrate and fecal N), suggesting that EA was more precise than KJ. In conclusion, drying of feces caused N losses regardless of drying methods. For urine and milk samples, if drying is necessary (i.e., EA), oven drying at 55°C can be used rather than lyophilization. The N content was greater in feeds, milk, and urine when determined with EA versus KJ. In addition, N content in acidified and undiluted urine at -20° changed and should be analyzed within 90 d of storage. The results in the current study, however, did not account for laboratory-to-laboratory variation., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Spray-dried plasma protein (SDP) compared with blood meal (BM) may contain various functional and active components that may benefit animal health. The objective of this experiment was to investigate the effects of feeding SDP or BM on production and blood profile in dairy cows during the transition and early-lactation periods. Seventy-two Holstein cows at 14 d before calving were used in a randomized block design. During the prepartum period, cows were fed a typical late-gestation diet containing BM (100 g/cow per day; 100BM, n = 24) or SDP (100 g/cow per day; 100SDP; n = 48). After calving, cows that were fed BM prepartum were fed a typical lactation diet formulated to provide 100 g/d of BM (100BM). Half the cows that were fed 100SDP prepartum were fed a lactation diet formulated to provide 100 g/d of SDP (100SDP; n = 24), and half were fed a diet formulated to provide 400 g/d of SDP (400SDP; n = 24) on a dry matter basis where SDP replaced BM (100SDP) or BM and soybean products (400SDP). All diets were balanced for crude protein concentration and metabolizable protein supply assuming BM and SDP were equal in rumen-degradable protein and rumen-undegradable protein. All data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC) as a randomized block design where contrasts were made for 100BM versus 100SDP for prepartum variables and 100BM versus 100SDP and 100SDP versus 400SDP for postpartum variables. Prepartum supplementation of SDP had no effect on plasma fatty acids and β-hydroxybutyrate (2 d before calving). Plasma fatty acids (255 ± 29 µEq/mL) and β-hydroxybutyrate (675 ± 70 µmol/L) at 8 and 14 d of lactation were not affected by SDP in the diet. Feeding SDP at 100 g/d compared with 100BM increased or tended to increase milk fat, protein, and lactose contents for 16 wk after parturition. Providing SDP at 400 g/d in the diet increased milk yield (42 vs. 39 kg/d), energy-corrected milk (44 vs. 41 kg/d), energy-corrected milk per kilogram of dry matter intake, and yields of milk fat (1.60 vs. 1.48 kg/d), protein (1.21 vs. 1.16 kg/d), and lactose compared with 100SDP. Body weight losses tended to be lower for 100SDP compared with 100BM without a difference between 100SDP and 400SDP. Plasma histidine concentration (d 14 of lactation) was lower for SDP compared with 100BM. In addition, plasma 1-methyl-l-histidine tended to be lower as inclusion rate of SDP increased. In conclusion, SDP at 400 g/d increased milk and milk component yields without an increase in feed intake. Studies evaluating effects of functional and active compounds in SDP on gut microbiome, gut health, and immune functions may be needed to determine mode of action., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Nutrient balance studies require measuring urine volume, and urinary excretion can be used to assess Mg bioavailability. A less laborious method than total collection of urine could make balance studies more feasible and expand the utility of using urinary Mg as an index of bioavailability, but the method needs to be accurate and sensitive. Sampling interval can affect accuracy because excretion must be at steady state. Two experiments were conducted to (1) determine whether urinary creatinine could be used to accurately estimate urinary output of nutrients markedly excreted via urine (N, K, Na, S, and Mg; experiment 1) and (2) determine the appropriate sampling schedule to evaluate Mg excretion after abrupt diet changes (experiment 2). Experiment 1 was originally designed to evaluate the interaction of monensin [0 vs. 14 mg of monensin/kg of dry matter (DM)] and Mg source (MgO vs. MgSO 4 ; total diet Mg: 0.36% of DM) under antagonism from increased dietary K (2.11% of DM) on urinary Mg excretion. Experiment 2 evaluated the interaction of Mg concentration (basal vs. supplemental MgO; total diet Mg: 0.20 vs. 0.42% of DM) and K (basal vs. supplemental K 2 CO 3 ; total diet K: 1.60 vs. 2.57% of DM) on urinary Mg excretion over time. Using 4-d composite samples from total collection of urine (n = 34 cow-periods), the average daily excretion of creatinine was similar to previous estimates (29.0 ± 1.16 mg of creatinine/kg of body weight) but was variable among cows (root mean squared error = 2,980 mg/d; 14% of mean). Treatment-average estimated excretion of urine and urinary N, K, Na, S, and Mg were similar to actual values; however, differences between actual and estimated values could be substantial for individual cows. Using the mean creatinine excretion per kilogram of body weight for all cows to estimate urine eliminates the lack of fit variance resulting in artificially low within-treatment variation for estimated urine volume. The standard error of the mean for estimated urine volume was 23% less (1.93 vs. 2.51) than that for actual urine production. This inflated the type I error rate, and, consequently, statistical inferences on N and K excretion differed when urine output was estimated rather than measured. The standard error of the mean for excretion of Mg calculated with actual or estimated urine production were almost identical (0.92 vs. 0.97); however, similar standard error of the mean was likely caused by differences in the covariance of urinary Mg concentration with estimated or actual urine output. Based on spot sampling (experiment 2), urinary Mg reached steady state by 2 d following an increase in dietary K regardless of Mg level, whereas excretion of urinary Mg following an increase in dietary Mg continued to increase through 7 d. Estimating nutrient excretion with urinary creatinine and body weight on average is accurate, but variance is likely underestimated. Knowing the time course of urinary Mg excretion will improve the value of using urinary Mg concentration to assess diet adequacy or Mg bioavailability., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Many nutrition models rely on summative equations to estimate feed and diet energy concentrations. These models partition feed into nutrient fractions and multiply the fractions by their estimated true digestibility, and the digestible mass provided by each fraction is then summed and converted to an energy value. Nonfiber carbohydrate (NFC) is used in many models. Although it behaves as a nutritionally uniform fraction, it is a heterogeneous mixture of components. To reduce the heterogeneity, we partitioned NFC into starch and residual organic matter (ROM), which is calculated as 100 - CP - LCFA - ash - starch - NDF, where crude protein (CP), long-chain fatty acids (LCFA), ash, starch, and neutral detergent fiber (NDF) are a percentage of DM. However, the true digestibility of ROM is unknown, and because NDF is contaminated with both ash and CP, those components are subtracted twice. The effect of ash and CP contamination of NDF on in vivo digestibility of NDF and ROM was evaluated using data from 2 total-collection digestibility experiments using lactating dairy cows. Digestibility of NDF was greater when it was corrected for ash and CP than without correction. Conversely, ROM apparent digestibility decreased when NDF was corrected for contamination. Although correcting for contamination statistically increased NDF digestibility, the effect was small; the average increase was 3.4%. The decrease in ROM digestibility was 7.4%. True digestibility of ROM is needed to incorporate ROM into summative equations. Data from multiple digestibility experiments (38 diets) using dairy cows were collated, and ROM concentrations were regressed on concentration of digestible ROM (ROM was calculated without adjusting for ash and CP contamination). The estimated true digestibility coefficient of ROM was 0.96 (SE = 0.021), and metabolic fecal ROM was 3.43 g/100 g of dry matter intake (SE = 0.30). Using a smaller data set (7 diets), estimated true digestibility of ROM when calculated using NDF corrected for ash and CP contamination was 0.87 (SE = 0.025), and metabolic fecal ROM was 3.76 g/100 g (SE = 0.60). Regardless of NDF method, ROM exhibited nutritional uniformity. The ROM fraction also had lower errors associated with the estimated true digestibility and its metabolic fecal fraction than did NFC. Therefore, ROM may result in more accurate estimates of available energy if integrated into models., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)