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This experiment investigated the variation in enteric methane production and associated gas exchange parameters, nutrient digestibility, rumen fermentation, and rumen microbiome when a range of silages based on different forage types (grass or clover), and different species within the 2 types, were fed as the sole feed to heifers. Three grass species (perennial ryegrass, festulolium, and tall fescue) and 2 clover species (red clover and white clover) were included. Perennial ryegrass was harvested at 2 maturity stages in the primary growth, white clover was harvested once in the primary growth, and 4 cuts of festulolium and tall fescue and 3 cuts of red clover were harvested during the growing season, giving 14 different silage batches in total. Sixteen Holstein heifers 16 to 21 mo old and 2 to 5 mo in pregnancy were fed the silages ad libitum as the sole feed in an incomplete crossover design. Each silage was fed to 4 heifers, except for the 2 perennial ryegrass silages, which were fed to 8 heifers; in total 64 observations. The CH4 production was measured for 3 d in respiration chambers. Heifers fed clover silages had higher dry matter intake (DMI) compared with heifers fed grass silages, and heifers fed tall fescue silages had the numerically the lowest DMI. Compared with grass silages, feeding clover silages led to higher crude protein digestibility but lower neutral detergent fiber (NDF) digestibility. Rumen pH was higher in heifers fed clover silages compared with those fed grass silages. Based on composition analysis, the rumen microbiota of the heifers clustered clearly according to forage type and species. More specifically, 7 of the 34 dominating rumen bacterial genus-level groups showed higher relative abundances for the clover silages, whereas 7 genus-level groups showed higher abundances for the grass silages. Methane yield was higher for heifers fed grass silages than for those fed clover silages when methane production was related to dry matter and digestible organic matter intake, whereas the opposite was seen when related to NDF digestion. The gross energy lost as methane (CH4 conversion factor, %) reduced from 7.5% to 6.7%, equivalent to an 11% reduction. The present study gives the outlines for choosing the optimal forage type and forage species with respect to nutrient digestibility and enteric methane emission in ruminants.

Little data is presently available on our ability to predict the combined effect of modifying diets with feeds rich in sugars or starch (ST) and rumen-degradable protein (RDP) on the performance of high-producing dairy cows. The objective of this study was to compare responses of 59 lactating Holstein cows to substitution of cane molasses (Mol) for dry corn grain (CG) at 3 levels of Mol and 2 levels of RDP (+RDP or -RDP) in a randomized complete block design with a 3 × 2 factorial arrangement of treatments. Also, lactation responses predicted by 2 nutritional models were compared with observed responses, with Mol composition entered so that nonnutritive materials in Mol were not counted as potentially digestible carbohydrate. We hypothesized that dry matter (DM) intake and milk fat percentage responses would increase with increasing Mol and would potentially be greater with +RDP. For evaluation of the nutritional models, we adopted the null hypothesis that observed and predicted lactation performance would not differ. Cows were individually fed a common diet during a 2-wk covariate period followed by 8 wk on experimental diets. Diets were formulated to be isonitrogenous and provide similar amounts of ST and water-soluble carbohydrates. Experimental diets contained, on a DM basis, 35% corn silage, 20% alfalfa silage, and 16.6% crude protein. The 0, 5.25, and 10.5% Mol diets respectively contained 19.0, 14.5, and 10.0% CG; 28, 25, and 22% ST; and 5.5, 8.5, and 11.5% water-soluble carbohydrates. At 10 wk on study, cows averaged 45.5 kg of energy-corrected milk (ECM). The DM intake (DMI), and yields of milk, milk protein, and ECM, and milk N/intake N declined linearly with increasing Mol. Differences among diets were not detected for milk fat yield and ECM/DMI. No RDP or interaction effects were detected for these measures. That milk production efficiency did not differ across diets suggests that DMI was a primary driver of performance. The similar ECM/DMI and maintenance of milk fat yield would not have been predicted based on Mol and CG composition but may relate to differences in fermentation rates and products. As explanation for these results, we hypothesize that more rapid ruminal evolution of volatile fatty acids post-ingestion with Mol compared with CG may have provided masses of acetate and butyrate in excess of existing energy and synthetic needs that were shunted to milk fat production, and of propionate that depressed intake. The 2001 Dairy National Research Council model and the Cornell Net Carbohydrate and Protein System 6.55 in Nutritional Dynamic System Professional (2021) estimates of metabolizable protein-allowable ECM underestimated actual ECM for +RDP diets by 4.5 and 2.3 kg, respectively, and came close or overestimated for -RDP diets by 0.25 and 5.0 kg, respectively. Prediction discrepancies suggest issues with valuation of dietary protein based on degradability. Improved understanding of factors mediating these results would likely enhance our ability to predict animal responses.

Questions regarding the balance between the contribution to human nutrition and the environmental impact of livestock food products rarely evaluate specific species or how to accomplish the recommended depopulation. The objective of this study was to assess current contributions of the US dairy industry to the supply of nutrients and environmental impact, characterize potential impacts of alternative land use for land previously used for crops for dairy cattle, and evaluate the impacts of these approaches on US dairy herd depopulation. We modeled 3 scenarios to reflect different sets of assumptions for how and why to remove dairy cattle from the US food production system coupled with 4 land-use strategies for the potential newly available land previously cropped for dairy feed. Scenarios also differed in assumptions of how to repurpose land previously used to grow grain for dairy cows. The current system provides sufficient fluid milk to meet the annual energy, protein, and calcium requirements of 71.2, 169, and 254 million people, respectively. Vitamins supplied by dairy products also make up a high proportion of total domestic supplies from foods, with dairy providing 39% of the vitamin A, 54% of the vitamin D, 47% of the riboflavin, 57% of the vitamin B12, and 29% of the choline available for human consumption in the United States. Retiring (maintaining animals without milk harvesting) dairy cattle under their current managemerit resulted in no change in absolute greenhouse gas emissions (GHGE) relative to the current production system. Both depopulation and retirement to pasture resulted in modest reductions (6.8-12.0%) in GHGE relative to the current agricultural system. Most dairy cow removal scenarios reduced availability of essential micronutrients such as a-linolenic acid, Ca, and vitamins A, D, B12, and choline. Those removal scenarios that did not reduce micronutrient availability also did not improve GHGE relative to the current production system. These results suggest that removal of dairy cattle to reduce GHGE without reducing the supply of the most limiting nutrients to the population would be difficult. Dairy Management Inc. (Rosemont, IL) Research was supported by Dairy Management Inc. (Rosemont, IL). The authors have not stated any conflicts of interest.

Ideal digesta markers used for feeding studies are inert, unabsorbable, and move with the digesta they are intended to mark. Both chromium (III) and cobalt (III) salts of EDTA (CrEDTA and CoEDTA, respectively) are used as markers of liquid digesta in dairy cattle research. A small portion is absorbed and excreted in urine, but the markers are assumed to remain unreactive and as inert salts in the digesta and animal. The degree to which these colored salts remain bound in solution can be estimated through spectrophotometric measurement at their wavelength (λ) of peak absorbance. The objective of this in vitro study was to evaluate whether CrEDTA and CoEDTA dissociate under reducing conditions that could be experienced in the rumen. In a completely randomized design with 2 replicate analytical runs and samples in duplicate within run, approximately 26 mg/L Cr from CrEDTA or Co from CoEDTA was incubated in a 26-mL reaction volume containing 20 mL of Goering and Van Soest medium without tryptone, 3 mL of CoEDTA or CrEDTA solutions, or water (reagent blanks), and 3.0 mL of a combination of distilled water with 0, 0.25, 0.50, 0.75, or 1.00 mL of reducing solution (RedSol). After incubation for 0.5 h at 39°C, absorbance was read at λ = 535, 465, and 560 nm, the peak λ for EDTA salts of Co(III), Co(II), and Cr(III), respectively. Mean reagent blank values were subtracted from CoEDTA and CrEDTA data. The absorbance data at peak λ were analyzed by marker in models that included RedSol with analytical run as a random variable. Contrasts were used to detect linear through quartic effects of RedSol. Samples with RedSol had redox potentials of -250 to -328 mV, which are within the range of reported ruminal measures. As RedSol increased, CoEDTA showed a linear decline of 75% in ABS at 535 nm and a quadratic 4-fold increase followed by a 60% decline at 465 nm. These responses indicate a reduction of Co(III) to Co(II) and subsequent dissociation of Co(II)EDTA. The absorbance of CrEDTA at 560 nm showed a tendency for an 8% linear decrease as RedSol increased. Wavescans from λ = 330 to 700 nm showed CrEDTA retaining its characteristic 2-peak pattern as RedSol increased, whereas CoEDTA curves deformed entirely. We conclude that CoEDTA is not a stable, inert digesta marker under reducing conditions achievable in the rumen and is therefore unsuitable for use in studies with ruminants. Reexamination of the suitability of available liquid digesta markers is advised.

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.

The objectives of this study were to determine if milk production efficiency (MPE) is altered by near-total exchange of ruminal contents between high- (HE) and low-MPE (LE) cows and to characterize ruminal bacterial community composition (BCC) before exchange and over time postexchange. Three pairs of ruminally cannulated, third-lactation cows were selected whose MPE (energy-corrected milk per unit of dry matter intake) differed over their first 2 lactations. Approximately 95% of ruminal contents were exchanged between cows within each pair. Ruminal pH and volatile fatty acid (VFA) profiles, along with BCC (characterized by sequencing of the variable 4 region of 16S rRNA genes), were assessed just before feeding on d -8, -7, -5, -4, -1, 1, 2, 3, 7, 10, 14, 21, 28, 35, 42, and 56, relative to the exchange date. High-MPE cows had higher total ruminal VFA concentrations, higher molar percentages of propionate and valerate, and lower molar percentages of acetate and butyrate than did LE cows, and re-established these differences 1 d after contents exchange. Across all LE cows, MPE increased during 7 d postexchange but declined thereafter. Two of the 3 HE cows displayed decreases in MPE following introduction of the ruminal contents from the corresponding LE cow, but MPE increased in the third HE cow, which was determined to be an outlier. For all 6 cows, both liquid- and solids-associated BCC differed between individuals within a pair before contents exchange. Upon exchange, BCC of both phases in all 3 pairs was more similar to that of the donor inoculum than to preexchange host BCC. For 5 of 6 cows, the solids-associated community returned within 10 d to more resemble the preexchange community of that host than that of the donor community. Individual variability before the exchange was greater in liquids than in solids, as was the variability in response of bacterial communities to the exchange. Individual cows varied in their response, but generally moved toward re-establishment of their preexchange communities by 10 d after contents exchange. By contrast, ruminal pH and VFA profiles returned to preexchange levels within 1 d. Despite the small number of cows studied, the data suggest an apparent role for the ruminal bacterial community as a determinant of MPE.

Millions of feed composition records generated annually by testing laboratories are valuable assets that can be used to benefit the animal nutrition community. However, it is challenging to manage, handle, and process feed composition data that originate from multiple sources, lack standardized feed names, and contain outliers. Efficient methods that consolidate and screen such data are needed to develop feed composition databases with accurate means and standard deviations (SD). Considering the interest of the animal science community in data management and the importance of feed composition tables for the animal industry, the objective was to develop a set of procedures to construct accurate feed composition tables from large data sets. A published statistical procedure, designed to screen feed composition data, was employed, modified, and programmed to operate using Python and SAS. The 2.76 million data received from 4 commercial feed testing laboratories were used to develop procedures and to construct tables summarizing feed composition. Briefly, feed names and nutrients across laboratories were standardized, and erroneous and duplicated records were removed. Histogram, univariate, and principal component analyses were used to identify and remove outliers having key nutrients outside of the mean ± 3.5 SD. Clustering procedures identified subgroups of feeds within a large data set. Aside from the clustering step that was programmed in Python to automatically execute in SAS, all steps were programmed and automatically conducted using Python followed by a manual evaluation of the resulting mean Pearson correlation matrices of clusters. The input data set contained 42, 94, 162, and 270 feeds from 4 laboratories and comprised 25 to 30 nutrients. The final database included 174 feeds and 1.48 million records. The developed procedures effectively classified by-products (e.g., distillers grains and solubles as low or high fat), forages (e.g., legume or grass-legume mixture by maturity), and oilseeds versus meal (e.g., soybeans as whole raw seeds vs. soybean meal expellers or solvent extracted) into distinct sub-populations. Results from these analyses suggest that the procedure can provide a robust tool to construct and update large feed data sets. This approach can also be used by commercial laboratories, feed manufacturers, animal producers, and other professionals to process feed composition data sets and update feed libraries.

An ideal digesta marker for use in feeding studies is inert, unabsorbable, and moves with the portion of the digesta it is intended to mark. Both chromium (III) and cobalt (III) salts of EDTA (CrEDTA and CoEDTA, respectively) have been used as digesta liquid markers in studies with dairy cattle. Although a small portion of these markers is known to be absorbed and excreted in urine, the markers are assumed to remain ionically bound in the digesta. The degree to which these salts remain bound in solution can be determined through spectrophotometric measurement at the wavelength (λ) of peak absorbance of these colored compounds. The objective of this study was to evaluate whether CrEDTA and CoEDTA dissociate during incubation in autoclaved, clarified rumen fluid (ACRF), as indicated by changes in absorbance. In a complete block design with separate replicated analytical runs, approximately 40 mg/L of Cr from CrEDTA or Co from CoEDTA were incubated in 2 separate preparations of ACRF from 2 lactating Holstein cows, in water (CrEDTA), or in MES buffer (CoEDTA), and appropriate reagent blanks. Solution pH were approximately 6.0. Samples were incubated for 24 h at 39°C with absorbance measurements recorded at 0, 1, 2, 4, 6, 22, and 24. The CrEDTA was measured at λ = 541 nm, CoEDTA at λ = 535 nm, and both were measured with wavescans of λ = 330 to 700 nm. At their peak λ, both CrEDTA in water and CoEDTA in MES buffer maintained absorbance values throughout the incubation, whereas, in ACRF, CrEDTA absorbance decreased by 9% at 0 h, and by up to 14% by 24 h; CoEDTA showed a gradual decline over time, with approximately 4% loss in absorbance by 24 h. Responses differed by ACRF preparation. Both markers in ACRF showed increases and decreases over time in absorbance at λ = 330 and 380 nm, though the changes were more marked for CrEDTA; markers not in ACRF showed no change in absorbance at these λ. Changes in the absorbance values at λ = 330 and 380 nm suggest that soluble phenolic compounds may be involved in the exchange of metals with EDTA, but that does not preclude involvement of other components in rumen fluid. Both CrEDTA and CoEDTA incubated in ACRF showed declines over time in absorbance at their peak λ, suggesting that the metals dissociated from EDTA. The apparent dissociation indicates that these liquid markers are not inert as had been assumed.

Glycogen is an α-glucan produced by rumen microbes from various feed carbohydrates. It may be digested ruminally or intestinally to provide nutrients. The physicochemical and enzymatic hydrolysis characteristics of microbial glycogen have not been described in detail, but do influence its conversion to absorbable nutrients in vivo, its nutritional comparability with plant starch sources, and its accurate analysis in vitro. The objectives of this study were to determine presence or absence of a gelatinization response and to describe enzymatic digestion characteristics of glycogen granules isolated from ruminal protozoa obtained from lactating dairy cows. Protozoal glycogen granules were determined to be 98.3% α-glucan. Granules displayed gelatinization, the breaking of hydrogen bonds between molecules or branches, at 65°C compared with purified wheat and corn starches, which initiated gelatinization at 50 and 65°C, respectively. Digestion of ungelatinized samples with amyloglucosidase for 2 h at 39°C showed approximately 3-fold greater hydrolysis to glucose for protozoal glycogen (25.2% of dry matter; DM) than for wheat (9.9% of DM) or corn (8.2% of DM) starches. Based on enzymatic digestion results, protozoal glycogen may be more readily digested than intact corn or wheat starches and should be gelatinized or the hydrogen bonds otherwise disrupted to allow more complete recovery in enzymatic analysis.

Postpartum dietary supplementation of gluconeogenic precursors may improve the plasma metabolite profile of dairy cows, reducing metabolic disorders and improving lactation performance. The objective of this trial was to examine the effects of supplementation with fermented ammoniated condensed whey (FACW) postpartum on lactation performance and on profile of plasma metabolites and hormones in transition dairy cows. Individually fed multiparous Holstein cows were blocked by calving date and randomly assigned to control (2.9% dry matter of diet as soybean meal; n = 20) or FACW (2.9% dry matter of diet as liquid GlucoBoost, Fermented Nutrition, Luxemburg, WI; n = 19) dietary treatments. Treatments were offered from 1 to 45 d in milk (DIM). Cows were milked twice a day. Dry matter intake and milk yield were recorded daily and averaged weekly. Individual milk samples from 2 consecutive milkings were obtained once a week for component analysis. Rumen fluid was collected (n = 3 cows/treatment) at 4 time points per day at 7 and 21 DIM. Blood samples were collected within 1 h before feeding time for metabolite analysis and hyperketonemia diagnosis. Supplementation of FACW improved feed efficiency relative to control; this effect may be partially explained by a marginally significant reduction in dry matter intake from wk 3 to 7 for FACW-supplemented cows with no detected FACW-driven changes in milk yield, milk protein yield, and milk energy output compared with control. Also, there was no evidence for differences in intake of net energy for lactation, efficiency of energy use, energy balance, or body weight or body condition score change from calving to 45 DIM between treatments. Supplementation of FACW shifted rumen measures toward greater molar proportions of propionate and butyrate, and lesser molar proportions of acetate and valerate. Cows supplemented with FACW had greater plasma glucose concentrations in the period from 3 to 7 DIM and greater plasma insulin concentrations compared with control. Plasma nonesterified fatty acid and β-hydroxybutyrate concentrations were decreased in cows supplemented with FACW compared with control cows in the period from 3 to 7 DIM. These findings indicate that FACW may have improved the plasma metabolite profile immediately postpartum in dairy cows. Additionally, supplementation of FACW resulted in improved feed efficiency as accessed by measures of milk output relative to feed intake.

Our knowledge of the role of carbohydrates in dairy cattle nutrition has advanced substantially in the 100 years of the publication of the Journal of Dairy Science. In this review, we trace the history of scientific investigation and discovery from crude fiber, nitrogen-free extract, and "unidentified factors" to our present analytical schemes and understanding of ruminal and whole-animal utilization and effects of dietary carbohydrates. Historically, advances in research and new feeding standards occurred in parallel with and fostered by new methods of analysis. The 100 years of research reviewed here has bequeathed to us an impressive legacy of information, which we will continue to grow.

The objective of this work was to leverage equations derived in a meta-analysis into an ensemble modeling system for estimating dietary physical and chemical characteristics required to maintain desired rumen conditions in lactating dairy cattle. Given the availability of data, responsiveness of ruminal pH to animal behaviors, and the chemical composition and physical form of the diet, mean ruminal pH was chosen as the primary rumen environment indicator. Physically effective fiber (peNDF) is defined as the fraction of neutral detergent fiber (NDF) that stimulates chewing activity and contributes to the floating mat of large particles in the rumen. The peNDF of feedstuffs is typically estimated by multiplying the NDF content by a particle size measure, resulting in an estimated index of effectiveness. We hypothesized that the utility of peNDF could be expanded and improved by dissociating NDF and particle size and considering other dietary factors, all integrated into a physically adjusted fiber system that can be used to estimate minimum particle sizes of TMR and diet compositions needed to maintain ruminal pH targets. Particle size measures of TMR were limited to those found with the Penn State particle separator (PSPS). Starting with specific diet characteristics, the system employed an ensemble of models that were integrated using a variable mixture of experts approach to generate more robust recommendations for the percentage of dietary DM material that should be retained on the 8-mm sieve of a PSPS. Additional continuous variables also integrated in the physically adjusted fiber system include the proportion of material (dry matter basis) retained on the 19- and 8-mm sieves of the PSPS, estimated mean particle size, the dietary concentrations of forage, forage NDF, starch, and NDF, and ruminally degraded starch and NDF. The system was able to predict that the minimum proportion of material (dry matter basis) retained on the 8-mm sieve should increase with decreasing forage NDF or dietary NDF. Additionally, the minimum proportion of dry matter material on the 8-mm sieve should increase with increasing dietary starch. Results of this study agreed with described interrelationships between the chemical and physical form of diets fed to dairy cows and quantified the links between NDF intake, diet particle size, and ruminal pH. Feeding recommendations can be interpolated from tables and figures included in this work.

Physically effective neutral detergent fiber (peNDF) is the fraction of neutral detergent fiber (NDF) that stimulates chewing activity and contributes to the floating mat of large particles in the rumen. Multiplying dietary NDF by particle size has been used as an estimate of peNDF. In re-evaluating the concept of peNDF, we compared the use of peNDF as dietary NDF x particle size with the use of individual NDF and particle size descriptors (physically adjusted NDF; paNDF) when used with other physical and chemical diet descriptors to predict dry matter (DM) intake (DMI), rumination time, and ruminal pH in lactating dairy cows. The purpose is to ultimately use these equations to estimate diet adequacy to maintain ruminal conditions. Each response variable had 8 models in a 2 (peNDF, paNDF) x 2 (diet, diet and ruminal factors) x 2 (DM, as fed basis) factorial arrangement. Particle size descriptors were those determined with the Penn State Particle Separator. Treatment means (n = 241) from 60 publications were used in backward elimination multiple regression to derive models of response variables. When available, peNDF terms entered equations. Models containing peNDF terms had similar or lower unadjusted concordance correlation coefficients (an indicator of similar or lower accuracy and precision) than did models without peNDF terms. The peNDF models for rumen pH did not differ substantially from paNDF models. This suggests that peNDF can account for some variation in ruminal pH; however, overt advantages of peNDF were not apparent. Significant variables that entered the models included estimated mean particle size; as fed or DM proportions retained on 19- and 8-mm sieves of the Penn State Particle Separator; DMI; dietary concentrations of forage; forage NDF; CP; starch; NDF; rumen-degraded starch and rumen-degraded NDF; and the interaction terms of starch x mean particle size, acid detergent fiber/NDF, and rumination time/DMI. Many dietary factors beyond particle size and NDF were identified as influencing the response variables. In conclusion, these results appear to justify the development of a modeling approach to integrate individual physical and chemical factors to predict effects on factors affecting rumen conditions. Agriculture and Food Research Initiative Competitive from the USDA National Institute of Food and Agriculture (Washington, DC) [2015-03656]; USDA Agricultural Research Service (Washington, DC) The authors thank Keri Rogan and Jared Judy for their efforts related to data entry. Funding for this project was provided by Agriculture and Food Research Initiative Competitive grant no. 2015-03656 from the USDA National Institute of Food and Agriculture (Washington, DC). Research was supported by state and federal funds appropriated to the University of Nebraska-Lincoln, Virginia Tech (Blacksburg), and The Ohio State University (Columbus) with funding from USDA Agricultural Research Service (Washington, DC). Public domain – authored by a U.S. government employee

In response to drought-induced forage shortages along with increased corn and soy prices, this study was conducted to evaluate lactation responses of dairy cows to lower-forage diets supplemented with forage substitutes. By-product feeds were used to completely replace corn grain and soybean feeds. Forty-eight late-lactation cows were assigned to 1 of 4 diets using a randomized complete block design with a 2-wk covariate period followed by a 4-wk experimental period. The covariate diet contained corn grain, soybean meal, and 61% forage. Experimental diets contained chopped wheat straw (WS)/sugar beet pulp at 0/12, 3/9, 6/6, or 9/3 percentages of diet dry matter (DM). Corn silage (20%), alfalfa silage (20%), pelleted corn gluten feed (25.5%), distillers grains (8%), whole cottonseed (5%), cane molasses/whey blend (7%), and vitamin and mineral mix with monensin (2.5%) comprised the rest of diet DM. The WS/sugar beet pulp diets averaged 16.5% crude protein, 35% neutral detergent fiber, and 11% starch (DM basis). Cows consuming the experimental diets maintained a 3.5% fat- and protein-corrected milk production (35.2 kg; standard deviation=5.6 kg) that was numerically similar to that measured in the covariate period (35.3 kg; standard deviation=5.0 kg). Intakes of DM and crude protein declined linearly as WS increased, whereas neutral detergent fiber intake increased linearly. Linear increases in time spent ruminating (from 409 to 502 min/d) and eating (from 156 to 223 min/d) were noted as WS inclusion increased. Yields of milk fat and 3.5% fat-and protein-corrected milk did not change as WS increased, but those of protein and lactose declined linearly. Phosphorous intakes were in excess of recommended levels and decreased linearly with increasing WS inclusion. Nutritional model predictions for multiparous cows were closest to actual performance for the National Research Council 2001 model when a metabolizable protein basis was used; primiparous cow performance was better predicted by energy-based predictions made with the National Research Council or Cornell Net Carbohydrate and Protein System models. Model predictions of performance showed a quadratic diet effect with increasing WS. Lactating dairy cows maintained production on low-forage diets that included forage substitutes, and in which by-product feeds fully replaced corn grain and soybean. However, longer-term studies are needed to evaluate animal performance and to improve model predictions of performance on these nontraditional diets.

Microbial fermentation of carbohy- drates in the hindgut of dairy cattle is responsible for 5 to 10% of total-tract carbohydrate digestion. When dietary, animal, or environmental factors contribute to abnormal, excessive flow of fermentable carbohydrates from the small intestine, hindgut acidosis can occur. Hindgut acidosis is characterized by increased rates of production of short-chain fatty acids including lactic acid, decreased digesta pH, and damage to gut epithe- lium as evidenced by the appearance of mucin casts in feces. Hindgut acidosis is more likely to occur in high-producing animals fed diets with relatively greater proportions of grains and lesser proportions of forage. In these animals, ruminal acidosis and poor selective retention of fermentable carbohydrates by the rumen will increase carbohydrate flow to the hindgut. In more severe situations, hindgut acidosis is characterized by an inflammatory response; the resulting breach of the barrier between animal and digesta may contribute to laminitis and other disorders. In a research setting, effects of increased hindgut fermentation have been evaluated using pulse-dose or continuous abomasal infusions of varying amounts of fermentable carbohy- drates. Continuous small-dose abomasal infusions of 1 kg/d of pectin or fructans into lactating cows resulted in decreased diet digestibility and decreased milk fat percentage without affecting fecal pH or VFA concen- trations. The decreased diet digestibility likely resulted from increased bulk in the digestive tract or from in- creased digesta passage rate, reducing exposure of the digesta to intestinal enzymes and epithelial absorptive surfaces. The same mechanism is proposed to explain the decreased milk fat percentage because only milk concentrations of long-chain fatty acids were decreased. Pulse-dose abomasal fructan infusions (1 g/kg of BW) into steers resulted in watery feces, decreased fecal pH, and increased fecal VFA concentrations, without caus- ing an inflammatory response. Daily 12-h abomasal in- fusions of a large dose of starch (~4 kg/d) have also induced hindgut acidosis as indicated by decreased fe- cal pH and watery feces. On the farm, watery or foamy feces or presence of mucin casts in feces may indicate hindgut acidosis. In summary, hindgut acidosis occurs because of relatively high rates of large intestinal fer- mentation, likely due to digestive dysfunction in other parts of the gut. A better understanding of the relation- ship of this disorder to other animal health disorders is needed.

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 x 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 x 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 30h 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 6h postfeeding was also influenced by the interaction of NFC x 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 NH(3) 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.

The concept of improving animal performance by going beyond simply meeting requirements and synchronizing ruminal availability of protein and energy has been with us for at least 3 decades. Although theoretically appealing, research and field results have not supported this approach to diet formulation. Why? Essential to successful ruminal synchrony is the ability to predict available amounts and fates of diverse substrates. The substrates come from varied sources; their efficiencies of use and yields of products are affected by inherent properties, interactions, transformations, and passage. However, substrate quality and availability in the rumen are affected only in part by diet. For example, NPN, true protein, and peptides are contributed by diet and intraruminal recycling, with additional endogenous NPN contributions by the cow. Changes in factors that alter the rate or extent of substrate fermentation, such as the rate of passage or ruminal pH, can alter nutrient yield from the rumen and must be accounted for in order for synchrony to work. Our ability to estimate ruminally available substrate is also challenged by normal variation in feed composition and imprecision in component and digestibility analyses. Current in vitro assays may not be adequate to accurately describe the digestibility of feed components in vivo in mixed diets. There are some indications that the amount or pattern of supply of fermentable carbohydrate has a greater impact on microbial production and efficiency than does the pattern of protein supply. Animal responses to modifications in the supply of true protein from the rumen may be masked if additional protein is oxidized by tissues or if AA from endogenous sources cover deficiencies. Animal factors, such as response to immune challenge and sustained damage to tissues, will also affect partitioning of nutrients for production and may alter an animal's response to changes in nutrient supply. With the array of factors internal and external to the diet that must be considered, "synchrony" implies a greater deliberate precision in diet manipulation than may be currently possible to effect. Perhaps we should consider balance. Within the rumen and cow, can we generate conditions so that needed substrates or nutrients are available from the diet or accessible from endogenous resources to meet requirements and enhance productivity and efficiency? This approach involves the whole animal, rather than only the rumen and feed we offer to the cow.

Selected vessel types and conditions used for in vitro fermentation were compared to evaluate their effects on determinations of neutral detergent fiber (NDF) digestibility (NDFD) in 2 replicate 48-h fermentations. Treatments included 50-mL polyethylene centrifuge tubes with gas-release valves (treatment 1); 50-mL polyethylene centrifuge tubes with continuous gassing with CO2 (treatment 2); 50-mL polyethylene centrifuge tubes sealed, oriented horizontally, and shaken continuously parallel to the long axis of the tube, with manual gas release (treatment 3); 125-mL Erlenmeyer flasks with continuous gassing with CO2 (treatment 4); and 125-mL serum vials sealed with stoppers and crimp seals with (treatment 5) or without (treatment 6) manual gas release. Goering and Van Soest medium and blended ruminal inoculum from 4 lactating cows were used. Substrates were alfalfa hay, corn silage, ryegrass hay, and soyhulls. Gas was released and measured in treatments 3 and 6 at 3.0, 5.5, 9.0, 11.5, 23.5, 29.5, and 47.5 h by using a syringe with a hypodermic needle. Vessels from each treatment were harvested at 0, 6, 12, 24, 30, and 48 h for NDF analysis, with NDFD calculated as 1 - [(residual NDF, g - residual NDF in fermentation blank, g)/sample NDF, g]. Medium pH did not decline below 6.3 for any treatment. Average values for NDFD for 24 through 48 h were 0.576, 0.639, 0.688, 0.668, 0.679, and 0.681 for treatments 1 through 6, respectively (standard error of the difference = 0.008). The lowest NDFD was noted for treatment 1, which differed from all other treatments; treatments 3, 4, 5, and 6 did not differ by treatment or by the interaction of treatment and substrate. Treatments 1 and 2 gave lower NDFD values than the other treatments, but these differences were not consistent and differed by substrate, with alfalfa showing the fewest differences among treatments and soyhulls the most. Net energy of lactation values for substrates, as predicted from differences in 48-h NDFD, were 7 to 15% lower for treatment 1 than for the average of all other treatments. Slopes of the gas production per gram of substrate dry matter curves differed between treatments 3 and 5. In conclusion, measured NDFD was altered by fermentation treatment, with polyethylene tubes + gas-release valves giving the lowest values. Consequently, NDFD values may not be comparable across fermentation methods, but the effect will vary among feedstuffs. The combination of methods used for sealing, gassing, or agitating vessels may have a greater impact on NDFD than does vessel type.

Effects of sucrose (Suc) concentration on fermentation kinetics and products were evaluated using 3 concentrations of Suc, with 1 concentration of isolated NDF from Bermudagrass fermented together in batch culture in vitro with rumen inoculum. Fixed amounts of medium and inoculum were the protein sources, so protein:Suc decreased with increasing Suc. Kinetics were calculated from gas production over 48 h in a randomized complete block design (n = 28), and product yield was evaluated with sampling every 4 h for 24 h in a split-split plot in time design (n = 84). Fermentation vial was the experimental unit. Increasing Suc increased the lag time of rapidly (P 1 indicate incorporation of C from the medium, likely from AA and peptides. The results support the premises that direct effects of Suc concentration and perhaps protein:Suc alter yields of fermentation products. That substrate concentration altered fermentation products and kinetics, possibly due to interactions with the run conditions, advises the clear definition of substrates and fermentation conditions to determine how the results integrate into our knowledge of ruminant nutrition.

Fructans are an important nonfiber carbohydrate in cool season grasses. Their fermentation by ruminal microbes is not well described, though such information is needed to understand their nutritional value to ruminants. Our objective was to compare kinetics and product formation of orchardgrass fructan (phlein; PHL) to other nonfiber carbohydrates when fermented in vitro with mixed or pure culture ruminal microbes. Studies were carried out as randomized complete block designs. All rates given are first-order rate constants. With mixed ruminal microbes, rate of substrate disappearance tended to be greater for glucose (GLC) than for PHL and chicory fructan (inulin; INU), which tended to differ from each other (0.74, 0.62, and 0.33 h(-1), respectively). Disappearance of GLC had almost no lag time (0.04 h), whereas the fructans had lags of 1.4h. The maximum microbial N accumulation, a proxy for cell growth, tended to be 20% greater for PHL and INU than for GLC. The N accumulation rate for GLC (1.31h(-1)) was greater than for PHL (0.75 h(-1)) and INU (0.26 h(-1)), which also differed. More microbial glycogen (+57%) was accumulated from GLC than from PHL, though accumulation rates did not differ (1.95 and 1.44 h(-1), respectively); little glycogen accumulated from INU. Rates of organic acid formation were 0.80, 0.28, and 0.80 h(-1) for GLC, INU, and PHL, respectively, with PHL tending to be greater than INU. Lactic acid production was more than 7-fold greater for GLC than for the fructans. The ratio of microbial cell carbon to organic acid carbon tended to be greater for PHL (0.90) and INU (0.86) than for GLC (0.69), indicating a greater yield of cell mass per amount of substrate fermented with fructans. Reduced microbial yield for GLC may relate to the greater glycogen production that requires ATP, and lactate production that yields less ATP; together, these processes could have reduced ATP available for cell growth. Acetate molar proportion was less for GLC than for fructans, and less for PHL than for INU. In studies with pure cultures, all microbes evaluated showed differences in specific growth rate constants (μ) for GLC, fructose, sucrose, maltose, and PHL. Selenomonas ruminantium and Streptococcus bovis showed the highest μ for PHL (0.55 and 0.67 h(-1), respectively), which were 50 to 60% of the μ achieved for GLC. The 10 other species tested had μ between 0.01 and 0.11h(-1) with PHL. Ruminal microbes use PHL differently than they do GLC or INU.

Alkaline treatment of gramineous crop residues can convert an abundant, minimally utilized, poorly digestible straw into a moderately digestible feedstuff. Given the volatile nature of grain prices, substitution of treated stover for grain was investigated with dairy cows to provide insights on ruminal and digestibility effects of a feed option that makes use of alternative, available resources. The objective of this study was to evaluate changes in diet digestibility and ruminal effects when increasing levels of calcium oxide-treated corn stover (CaOSt) were substituted for corn grain in diets of lactating cows. Mature corn stover was treated with calcium oxide at a level of 50g∙kg(-1) dry matter (DM), brought up to a moisture content of 50% following bale grinding, and stored anaerobically at ambient temperatures for greater than 60d before the feeding experiment. Eight ruminally cannulated Holstein cows averaging 686kg of body weight and 35kg of milk∙d(-1) were enrolled in a replicated 4×4 Latin square, where CaOSt replaced corn grain on a DM basis in the ration at rates of 0, 40, 80, and 120g∙kg(-1) DM. All reported significant responses were linear. The DM intake declined by approximately 1kg per 4% increase in CaOSt inclusion. With increasing replacement of corn grain, dietary neutral detergent fiber (NDF) concentration increased. However, rumen NDF turnover, NDF digestibility, NDF passage rate, and digestion rate of potentially digestible NDF were unaffected by increasing CaOSt inclusion. Total-tract organic matter digestibility declined by 5 percentage units over the range of treatments, approximately 1.5 units per 4-percentage-unit substitution of CaOSt for grain. With increasing CaOSt, the molar proportions of butyrate and valerate declined, whereas the lowest detected ruminal pH increased from 5.83 to 5.94. Milk, fat, and protein yields declined as CaOSt increased and DM intake declined with the result that net energy in milk declined by approximately 1 Mcal per 4% increase in CaOSt. Time spent ruminating (min∙kg(-1) DM intake) increased with increasing CaOSt, though total minutes per day were unaffected. These insights on the effect of substitution of treated corn stover for corn grain may be used to predict the effect on nutrient supply to the cow over a range of substitution levels. The acceptability of the effect will depend on the economics of milk production and availabilities of feedstuffs.

Evaluation of physical, chemical, and enzymatic hydrolysis characteristics of protozoal glycogen is best performed on a pure substrate to avoid interference from other cell components. A method for isolating protozoal glycogen granules without use of detergents or other potentially contaminating chemicals was developed. Rumen inoculum was incubated anerobically in vitro with glucose. Glycogen-laden protozoa produced in the fermentation, primarily isotrichids, were allowed to sediment in a separatory funnel and were dispensed. The protozoa were processed through repeated centrifugations and sonication to isolate glycogen granules largely free of feed and cellular debris. The final water-insoluble lyophilized product analyzed as 98.3% α-glucan with very rare starch granules and 1.9% protein. Observed losses of glycogen granules during the clean-up process indicate that this procedure should not be used for quantitative assessment of protozoal glycogen from fermentations. Further optimization of this procedure to enhance the amount of glycogen obtained per fermentation may be possible.

Volatile fatty acid concentrations ([VFA], mM) have long been used to assess the effect of dietary treatments on ruminal fermentation in vivo. However, discrepancies in statistical results between [VFA] and VFA pool size (VFAmol) possibly related to ruminal digesta liquid amount (LIQ, kg) indicate potential issues with the use of [VFA]. We investigated relationships among [VFA], VFAmol, and LIQ measured 2 h postfeeding using individual lactating cow data (n=175) from 7 separate feeding studies. Regression analyses were performed using mixed models with "study" as a discrete random variable. The mean across studies and average range of values within studies, respectively, were 151 and 75 for [VFA], 11.2 and 9.8 for VFAmol, 73.3 and 41.0 for LIQ, and 289 and 83 mmol/kg for rumen fluid osmolality. Liquid amount changed with VFAmol (3.76 VFAmol+31.2; average within-study R2=0.69), but the relationship was weak between [VFA] and LIQ (0.524 LIQ+112.8; average within-study R2=0.12). The relationship between LIQ and VFAmol was likely a function of the osmotic gradient between rumen liquid and blood. The VFA are a major ruminal solute; VFAmol amounts can affect water flux in the rumen as similar tonicities of rumen fluid and blood are maintained. This also has a damping effect on ruminal solute concentration, creating the weak relationship between [VFA] and LIQ. Within studies, similar [VFA] were found in LIQ differing by 30 kg or more. The difference between minimum and maximum LIQ within cow within study was 12.7 kg (standard deviation=7.1), so inclusion of "cow" in analyses did not correct for the variation in LIQ. To allow valid comparisons of experimental treatments, responses must be on an equivalent basis; concentrations in different LIQ are not on an equivalent basis and so are not valid to use for comparing treatment effects. The [VFA] changed with VFAmol (5.80 VFAmol+86.3; average within-study R2=0.56). However, the ratio of [VFA] to VFAmol ranged from 9.0 to 24.1 as a function of 1,000/LIQ; this reflects the inherent calculated relationship among the variables. The varying relationship of [VFA] to VFAmol further indicates that [VFA] is not an appropriate measure to evaluate the progress or effect of treatments on ruminal fermentation. Predictions of LIQ and VFAmol using cow and ruminal measures were insufficiently precise to be used in research. Previously drawn conclusions based on [VFA] need to be reevaluated, and alternate evaluations for in vivo ruminal fermentation are needed.

An important objective is to identify nutrients or dietary factors that are most critical for advancing our knowledge of, and improving our ability to predict, milk protein production. The Dairy NRC (2001) model is sensitive to prediction of microbial protein synthesis, which is among the most important component of models integrating requirement and corresponding supply of metabolizable protein or amino acids. There are a variety of important considerations when assessing appropriate use of microbial marker methodology. Statistical formulas and examples are included to document and explain limitations in using a calibration equation from a source publication to predict duodenal flow of purine bases from measured urinary purine derivatives in a future study, and an improved approach was derived. Sources of specific carbohydrate rumen-degraded protein components probably explain microbial interactions and differences among studies. Changes in microbial populations might explain the variation in ruminal outflow of biohydrogenation intermediates that modify milk fat secretion. Finally, microbial protein synthesis can be better integrated with the production of volatile fatty acids, which do not necessarily reflect volatile fatty acid molar proportions in the rumen. The gut and splanchnic tissues metabolize varying amounts of volatile fatty acids, and propionate has important hormonal responses influencing milk protein percentage. Integration of ruminal metabolism with that in the mammary and peripheral tissues can be improved to increase the efficiency of conversion of dietary nutrients into milk components for more efficient milk production with decreased environmental impact.

Water is a critical nutrient for dairy cows, with intake varying with environment, production, and diet. However, little work has evaluated the effects of water intake on rumen parameters. Using dietary potassium carbonate (K2CO3) as a K supplement to increase water intake, the objective of this study was to evaluate the effect of K2CO3 supplementation on water intake and on rumen parameters of lactating dairy cows. Nine ruminally cannulated, late-lactation Holstein cows (207 ± 12 d in milk) were randomly assigned to 1 of 3 treatments in a replicated 3 × 3 Latin square design with 18-d periods. Dietary treatments (on a dry matter basis) were no added K2CO3 (baseline dietary K levels of 1.67% dietary K), 0.75% added dietary K, and 1.5% added dietary K. Cows were offered treatment diets for a 14-d adaption period followed by a 4-d collection period. Ruminal total, liquid, and dry matter digesta weights were determined by total rumen evacuations conducted 2 h after feeding on d 4 of the collection period. Rumen fluid samples were collected to determine pH, volatile fatty acids, and NH3 concentrations, and Co-EDTA was used to determine fractional liquid passage rate. Milk samples were collected twice daily during the collection period. Milk, milk fat, and protein yields showed quadratic responses with greatest yields for the 0.75% added dietary K treatment. Dry matter intake showed a quadratic response with 21.8 kg/d for the 0.75% added dietary K treatment and 20.4 and 20.5 kg/d for control and the 1.5% added dietary K treatment, respectively. Water intake increased linearly with increasing K2CO3 supplementation (102.4, 118.4, and 129.3 L/d) as did ruminal fractional liquid passage rate in the earlier hours after feeding (0.118, 0.135, and 0.141 per hour). Total and wet weights of rumen contents declined linearly and dry weight tended to decline linearly as dietary K2CO3 increased, suggesting that the increasing water intake and fractional liquid passage rate with increasing K2CO3 increased the overall ruminal turnover rate. Ruminal ammonia concentrations declined linearly and pH increased linearly as K supplementation increased. As a molar percentage of total volatile fatty acids, acetate increased linearly as dietary K increased, though propionate declined. Increasing dietary K2CO3 and total K in the diets of lactating dairy cows increased water consumption and modified ruminal measures in ways suggesting that both liquid and total ruminal turnover were increased as both water and K intake increased.

The objective of this study was to determine the effect of substituting corn bran (CB) for dried ground corn grain (CG) in the nonforage portion of high-forage (HF) and low-forage (LF) diets. Twelve multiparous and 12 primiparous Holsteins were assigned to 4 diets using six 4× Latin squares with 3-wk periods. Forage was 64 or 38% of the total mixed ration (% of dry matter). On a dry matter basis, the HFCG diet had 20% CG, the LFCG diet had 39% CG, the HFCB diet had 19% CB, and the LFCB diet had 38% CB. Digestible organic matter intake (OMI) and milk energy yield were lower for CB compared with CG within forage level. Digestible OMI was greater (1.9kg/d) for the LFCG compared with the HFCG treatment. When CB replaced forage (LFCB vs. HFCB), digestible OMI was not different but milk energy yield was greater with the LFCB diet. The LFCG diet supported the greatest milk, milk protein, and milk energy yield. Decreased concentration of milk protein and increased concentration of milk urea nitrogen when feeding CB compared with CG suggests that lack of fermentable energy in the CB diets may have limited rumen microbial protein synthesis. Total substitution of CG with CB in the nonforage portion did not support maximum milk production, even when forage was reduced at the same time (HFCG vs. LFCB). Predicted neutral detergent fiber (NDF) digestibility at 1 times maintenance, based on chemical analysis of the individual feeds, was 22 percentage units greater for CB than for the forage mix (68.9 vs. 46.9%). In vitro NDF digestibility (30h) was 19.4 percentage units greater for CB than for the forage mix (68.9 vs. 49.5%). However, in vivo NDF digestibility of the diet when CB replaced forage (HFCB vs. LFCB) was similar (44.1 vs. 44.5%). Similarly, predicted total digestible nutrients at the production level of intake, based on chemical analysis, were greater for the CB treatments and lower for the CG treatments than those observed in vivo.

In vitro neutral detergent fiber (NDF) digestibility (NDFD) is an empirical measurement of fiber fermentability by rumen microbes. Variation is inherent in all assays and may be increased as multiple steps or differing procedures are used to assess an empirical measure. The main objective of this study was to evaluate variability within and among laboratories of 30-h NDFD values analyzed in repeated runs. Subsamples of alfalfa (n=4), corn forage (n=5), and grass (n=5) ground to pass a 6-mm screen passed a test for homogeneity. The 14 samples were sent to 10 laboratories on 3 occasions over 12 mo. Laboratories ground the samples and ran 1 to 3 replicates of each sample within fermentation run and analyzed 2 or 3 sets of samples. Laboratories used 1 of 2 NDFD procedures: 8 labs used procedures related to the 1970 Goering and Van Soest (GVS) procedure using fermentation vessels or filter bags, and 2 used a procedure with preincubated inoculum (PInc). Means and standard deviations (SD) of sample replicates within run within laboratory (lab) were evaluated with a statistical model that included lab, run within lab, sample, and lab × sample interaction as factors. All factors affected mean values for 30-h NDFD. The lab × sample effect suggests against a simple lab bias in mean values. The SD ranged from 0.49 to 3.37% NDFD and were influenced by lab and run within lab. The GVS procedure gave greater NDFD values than PInc, with an average difference across all samples of 17% NDFD. Because of the differences between GVS and PInc, we recommend using results in contexts appropriate to each procedure. The 95% probability limits for within-lab repeatability and among-lab reproducibility for GVS mean values were 10.2 and 13.4%, respectively. These percentages describe the span of the range around the mean into which 95% of analytical results for a sample fall for values generated within a lab and among labs. This degree of precision was supported in that the average maximum difference between samples that were not declared different by means separation was 4.4% NDFD. Although the values did not have great precision, GVS labs were able to reliably rank sample data in order of 30-h NDFD (Spearman correlation coefficient = 0.93) with 80% of the rankings correct or off by only 1 ranking. A relative ranking system for NDFD could reduce the effect of within- and among-lab variation in numeric values. Such a system could give a more accurate portrayal of the comparative values of samples than current numeric values imply.

The goal of this in vitro study was to determine the influence of isotrichid protozoa (IP) on the conversion of glucose (Glc) to glycogen (Glyc) and transformation of Glc into fermentation products. Treatments were ruminal inoculum mechanically processed (blended) to destroy IP (B+, verified microscopically) or not mechanically processed (B-). Accumulated microbial Glyc was measured at 3h of fermentation with (L+; protozoa+bacteria) or without (L- predominantly protozoa) lysis of bacterial cells in the fermentation solids with 0.2 N NaOH. Two 3-h in vitro fermentations were performed using Goering-Van Soest medium in batch culture vessels supplemented with 78.75 mg of Glc/vessel in a 26.5-mL liquid volume. Rumen inoculum from 2 cannulated cows was filtered through cheesecloth, combined, and maintained under CO(2) for all procedures. At 3h, 0.63 and 0.38 mg of Glc remained in B- and B+. Net microbial Glyc accumulation (and Glc in Glyc as % of added Glc) detected at 3h of fermentation were 3.32 (4.69%), -1.42 (-2.01%), 6.45 (9.10%), and 3.65 (5.15%) mg for B-L-, B+L-, B-L+ and B+L+, respectively. Treatments B+ and L+ gave lower Glyc values than B- and L-, respectively. Treatment B+L- demonstrated net utilization of α-glucan contributed by inoculum with no net Glyc production. With destruction of IP, total Glyc accumulation declined by 44%, but estimated bacterial Glyc increased. Microbial accumulation of N increased 17.7% and calculated CH(4) production decreased 24.7% in B+ compared with B-, but accumulation of C in microbes, production of organic acids or C in organic acids, calculated CO(2), and carbohydrates in cell-free medium did not differ between B+ and B-. Given the short 3-h timeframe, increased N accumulation in B+ was attributed to decreased Glyc sequestration by IP rather than decreased predation on bacteria. After correction for estimates of C from AA and peptides utilized by microbes, 15% of substrate Glc C could not be accounted for in measured products in B+ or B-. Approximately 30% of substrate Glc was consumed by energetic costs associated with Glc transport and Glyc synthesis. The substantial accumulation of Glyc and changes in microbial N and Glyc accumulation related to presence of IP suggest that these factors should be considered in predicting profiles and amounts of microbial products and yield of nutrients to the cow as related to utilization of glucose. Determination of applicability of these findings to other soluble carbohydrates could be useful.

Methods for processing feedstuffs before analysis can affect analytical results. Effects of drying temperature (corn silage), preservation method (corn grain), and grinding method (corn silage and grain) on starch analysis values were evaluated. Corn silage samples dried at 55 or 105 degrees C and grain samples dried at 55 degrees C were ground to pass the 1-mm screen of an abrasion mill or cutting mill and analyzed for free glucose and starch corrected for free glucose. Starch analyses were performed in triplicate to assess the effect of treatment on precision of starch determination. Drying at 105 degrees C decreased free glucose and tended to decrease starch detected in silage. Decreased free glucose and starch values in silages dried at 105 degrees C may have been caused by the destruction of glucose and production of Maillard products through nonenzymatic browning. Maillard products with reducing activity could potentially interfere with the glucose oxidase-peroxidase glucose detection method used. Compared with the cutting mill, grinding samples through the abrasion mill increased the precision of starch measures in silage, likely due to the effect of the finer particle size produced by the abrasion mill allowing more accurate subsampling of a more homogeneous matrix. Starch values were greater for grain ground with an abrasion mill than with a cutting mill, with the difference greater for dry-rolled than for high-moisture corn. For starch analysis of corn silage and corn grain, drying at lower temperatures (55 degrees C) in forced-air ovens and grinding through the 1-mm screen of an abrasion mill or its equivalent is recommended.

Soyhulls (original form), neutral detergent and acid detergent fiber extracted from soyhulls were characterized by rates of dry matter disappearance by in situ and in vitro enzyme procedures. Enzyme combinations were cellulase, pectinase, cellulase – hemicellulase, cellulase – pectinase, and cellulase – hemicellulase – pectinase. Incubation times for both in situ and in vitro procedures were 0, 1, 2, 6, 8, 12, and 24 h. Two additional independent time sequences of soyhulls used in situ and cellulase procedures for extended times of 36, 48 h and 36, 48, 60, and 72 h. In situ rates of dry matter disappearance were similar to in vitro enzyme systems with cellulase or cellulase – hemicellulase for original hulls at 24 h. In situ neutral detergent fiber disappearance was similar to cellulase, and acid detergent in situ disappearance similar to cellulase – pectinase. Addition of pectinase to cellulase or cellulase – hemicellulase increased dry matter disappearance for original and neutral detergent hulls but not acid detergent hulls. In situ rates of dry matter disappearance for original soyhulls increased with incubation time at 72 h compared to 24 and 48 h, emphasizing the importance of extended incubation for this technique. In vitro rates for cellulase decreased with incubation beyond 24 h, suggesting caution in interpretation of rate for in vitro enzymatic systems using extended intervals.