The diurnal patterns of ruminal enzymatic activity and in vitro digestibility of starch, neutral detergent fiber, and protein.

The objective of this study was to determine whether diurnal patterns in starch, neutral detergent fiber (NDF) and protein digestibilities and amylolytic, fibrolytic, and proteolytic activities exist in dairy cows. Rumen fluid was collected from 4 ruminally cannulated Holstein dairy cows before the morning feeding and subsequently every 4 h for a 24-h period. Two of the cows were restricted from feed for 8 h overnight, and the other 2 continued to receive their feed ad libitum, to isolate and quantify the effects of changes in feeding behavior at night. After 2 runs the cows were crossed over between night feeding treatments. Rumen fluid was analyzed for enzymatic activity and in vitro starch, NDF, and nitrogen digestibility. Circadian rhythm analyses of enzymatic activity and in vitro digestibility were conducted by fitting the linear form of a cosine function with a 24-h period. Patterns were observed in activity for amylase, lichenase, endoglucanase, and xylanase, with the highest activities observed at the time points subsequent to milking and feed delivery. Protease activity was unaffected by either feeding treatment or possible feeding behavior. When fitted to a cosine function, all the parameters tested followed a daily pattern that was sensitive to the overnight availability of feed, although the parameters responded differently to the feeding treatment. The patterns displayed by in vitro digestibility results of starch, NDF, and nitrogen, across the various fluid collection time points, were highly variable. The time at peak (acrophase) observed in the enzymatic analysis did not correspond to those observed in the in vitro analysis. These results suggest that different interpretations should be given to enzymatic activities and in vitro digestibility values, and the time of rumen fluid collection relative to feeding time should be considered and reported when rumen fluid is used for research or commercial purposes. Maximum digestibility appears in fact to be reached around 4 to 5 h after the main ration delivery for NDF and starch and around ration delivery for protein.

Development of a mathematical model to predict pool sizes and rates of digestion of 2 pools of digestible neutral detergent fiber and an undigested neutral detergent fiber fraction within various forages.

The digestion of neutral detergent fiber treated with amylase and sodium sulfite and ash corrected (aNDFom) has been described as a single digestible pool and a fraction undigested in ruminants. Further, most models that predict rate and extent of digestion of aNDFom in the rumen assume first-order processes, in which the rates of digestion and passage are proportional to the pool size of aNDFom consumed and digested. Data exist demonstrating that the undigested fraction of aNDFom is not well described by a fixed coefficient and varies by maturity and agronomic growing conditions of the plant. Thus, our objective was to improve the prediction of digestible aNDFom (pdNDF) and to quantify, using a minimum number of fermentation time points, 2 pools of digestible aNDFom, pdNDF1 and pdNDF2, and their respective rates. Based on fermentations from 0 to 240 h among 34 forages (grasses, conventional and brown midrib corn silages, and alfalfas), 3 pools were described by aNDFomt = pdNDF1 × e-k1(t-L) + pdNDF2 × e-k2(t-L) + uNDF, where aNDFomt is the residue at time t; L is the lag; k1 is the rate of digestion of pdNDF1; k2 is the rate of digestion of pdNDF2; and uNDF the unavailable NDF on an aNDFom basis. A nonlinear estimation allowed the computation of the pool size and respective digestion rates. Using 3 time points from the digestion curve, 30, 120, and 240 h, as the fermentation endpoints to represent uNDF, we optimized the same model in Vensim (Ventana Simulation Environment; Ventana Systems Inc., Belmont, MA) to obtain rates and pool sizes of aNDFom. In addition, the same optimization was also performed with 2 timepoints and a forage type-specific range for uNDF. Parameters (with and without uNDF) obtained per forage using Vensim were compared by fitting kinetics data from the nonlinear calculations, using coefficients of determination and residual mean squares at convergence for ranking purposes for the whole equation and mean squared prediction errors for specific parameters. The highest coefficient of determination (0.98) and lowest mean square prediction error [0.0927 (NDF-1)2] were obtained when using 48, 120, and 240 h of aNDFom residues or when using 30 and 120 h and a range for the forage-specific uNDFom. Correlations were in all cases consistently high for all kinetic parameters, ranging from 0.76 to 0.99. Results demonstrated that an adequate description of the heterogeneity of aNDFom disappearance was possible without multiple fermentation time points. The equation was fit to all data generated; however, because of the variable nature of pool sizes and rates, forage-specific equations should be developed for better estimations of the forage specific pool sizes and uNDF estimation. This study further describes the heterogeneous nature of aNDFom disappearance and provides an approach for estimating the individual pool sizes and rates of digestion for application for diet formulation.

Development of an in vitro method to determine rumen undigested aNDFom for use in feed evaluation.

A portion of the forage cell wall, defined as neutral detergent fiber (NDF), is indigestible to anaerobic microbial digestion in ruminants. This fraction has been characterized by surface area relationships between acid detergent lignin, but recently, data have been published describing the dynamic nature of this relationship. In situ approaches have been described to estimate indigestible NDF, recovering the undigested NDF after long-term fermentations (uNDF). To be applicable to nutritionists and diet formulation, determining uNDF needs to be conducted in a commercial laboratory similar to other routine analyses of forage chemistry. A series of studies were conducted to evaluate an in vitro approach, to describe uNDF, which is repeatable and adaptable for routine feed evaluation. One hundred and two forages of several species were analyzed for NDF, acid detergent lignin, and uNDF. The uNDF was estimated by several approaches involving long-term fermentations and filtration steps to evaluate the length of time necessary to exhaust the digestible NDF and a filtration method necessary to maintain sample integrity by ensuring low sample loss and uniform recovery with residues from long-term in vitro fermentation. To determine uNDF, in vitro fermentations were conducted on 0.50 or 0.75 g of dry matter samples, in triplicate, at multiple time points up to 504 h and initially used Gooch crucibles with Celite (Thermo Fisher Scientific, Waltham, MA) as a filtering aid. The final method utilized a 1.5-µm pore size glass microfiber filter, which allowed for increased repeatability and improved sample recovery (lowest standard deviation). In this study, in vitro fermentations of 240 h were adequate to characterize and identify uNDF, which was repeatable among conventional forages provided the samples, after NDF analyses, were filtered through the same glass fiber filter. This approach could be adapted by commercial laboratories and would provide opportunities to develop near-infrared reflectance spectroscopy equations and calibrations.

Effect of lignin linkages with other plant cell wall components on in vitro and in vivo neutral detergent fiber digestibility and rate of digestion of grass forages.

The objective of this study was to correlate in vitro and in vivo neutral detergent fiber (NDF) digestibility (NDFD) with the chemical composition of forages and specific chemical linkages, primarily ester- and ether-linked para-coumaric (pCA) and ferulic acids (FA) in forages fed to dairy cattle. The content of acid detergent lignin (ADL) and its relationship with NDF does not fully explain the observed variability in NDFD. The ferulic and p-coumaric acid linkages between ADL and cell wall polysaccharides, rather than the amount of ADL, might be a better predictor of NDFD. Twenty-three forages, including conventional and brown midrib corn silages and grasses at various stages of maturity were incubated in vitro for measurement of 24-h and 96-h NDFD. Undigested and digested residues were analyzed for NDF, acid detergent fiber (ADF), ADL, and Klason lignin (KL); ester- and ether-linked pCA and FA were determined in these fractions. To determine whether in vitro observations of ester- and ether-linked pCA and FA and digestibility were similar to in vivo observations, 3 corn silages selected for digestibility were fed to 6 ruminally fistulated cows for 3 wk in 3 iso-NDF diets. Intact samples and NDF and ADF residues of diet, rumen, and feces were analyzed for ester- and ether-linked pCA and FA. From the in vitro study, the phenolic acid content (total pCA and FA) was highest for corn silages, and overall the content of ester- and ether-linked pCA and FA in both NDF and ADF residues were correlated with NDF digestibility parameters, reflecting the competitive effect of these linkages on digestibility. Also, Klason lignin and ADL were negatively correlated with ether-linked ferulic acid on an NDF basis. Overall, esterified FA and esterified pCA were negatively correlated with all of the measured fiber fractions on both a dry matter and an NDF basis. The lignin content of the plant residues and chemical linkages explained most of the variation in both rate and extent of NDF digestion but not uniformly among forages, ranging from 56 to 99%. The results from the in vivo study were similar to the in vitro data, demonstrating the highest total-tract aNDF digestibility (70%; NDF analysis conducted with α-amylase and sodium sulfite) for cows fed the corn silage with the lowest ester- and ether-linked pCA content in the NDF fraction. In this study, digestibility of forage fiber was influenced by the linkages among lignin and the carbohydrate moieties, which vary by hybrid and species and most likely vary by the agronomic conditions under which the plant was grown.

The Cornell Net Carbohydrate and Protein System: Updates to the model and evaluation of version 6.5.

New laboratory and animal sampling methods and data have been generated over the last 10 yr that had the potential to improve the predictions for energy, protein, and AA supply and requirements in the Cornell Net Carbohydrate and Protein System (CNCPS). The objectives of this study were to describe updates to the CNCPS and evaluate model performance against both literature and on-farm data. The changes to the feed library were significant and are reported in a separate manuscript. Degradation rates of protein and carbohydrate fractions were adjusted according to new fractionation schemes, and corresponding changes to equations used to calculate rumen outflows and postrumen digestion were presented. In response to the feed-library changes and an increased supply of essential AA because of updated contents of AA, a combined efficiency of use was adopted in place of separate calculations for maintenance and lactation to better represent the biology of the cow. Four different data sets were developed to evaluate Lys and Met requirements, rumen N balance, and milk yield predictions. In total 99 peer-reviewed studies with 389 treatments and 15 regional farms with 50 different diets were included. The broken-line model with plateau was used to identify the concentration of Lys and Met that maximizes milk protein yield and content. Results suggested concentrations of 7.00 and 2.60% of metabolizable protein (MP) for Lys and Met, respectively, for maximal protein yield and 6.77 and 2.85% of MP for Lys and Met, respectively, for maximal protein content. Updated AA concentrations were numerically higher for Lys and 11 to 18% higher for Met compared with CNCPS v6.0, and this is attributed to the increased content of Met and Lys in feeds that were previously incorrectly analyzed and described. The prediction of postruminal flows of N and milk yield were evaluated using the correlation coefficient from the BLUP (R(2)BLUP) procedure or model predictions (R(2)MDP) and the concordance correlation coefficient. The accuracy and precision of rumen-degradable N and undegradable N and bacterial N flows were improved with reduced bias. The CNCPS v6.5 predicted accurate and precise milk yield according to the first-limiting nutrient (MP or metabolizable energy) with a R(2)BLUP=0.97, R(2)MDP=0.78, and concordance correlation coefficient=0.83. Furthermore, MP-allowable milk was predicted with greater precision than metabolizable energy-allowable milk (R(2)MDP=0.82 and 0.76, respectively, for MP and metabolizable energy). Results suggest a significant improvement of the model, especially under conditions of MP limitation.

Preweaning milk replacer intake and effects on long-term productivity of dairy calves.

The preweaning management of dairy calves over the last 30 yr has focused on mortality, early weaning, and rumen development. Recent studies suggest that nutrient intake from milk or milk replacer during the preweaning period alters the phenotypic expression for milk yield. The objective of this study was to investigate the relationship between nutrient intake from milk replacer and pre- and postweaning growth rate with lactation performance in the Cornell dairy herd and a commercial dairy farm. The analysis was conducted using traditional 305-d first-lactation milk yield and residual lactation yield estimates from a test-day model (TDM) to analyze the lactation records over multiple lactations. The overall objective of the calf nutrition program in both herds was to double the birth weight of calves by weaning through increased milk replacer and starter intake. First-lactation 305-d milk yield and residuals from the TDM were generated from 1,244 and 624 heifers from the Cornell herd and from the commercial farm, respectively. The TDM was used to generate lactation residuals after accounting for the effects of test day, calving season, days in milk, days pregnant, lactation number, and year. In addition, lactation residuals were generated for cattle with multiple lactations to determine if the effect of preweaning nutrition could be associated with lifetime milk yield. Factors such as preweaning average daily gain (ADG), energy intake from milk replacer as a multiple of maintenance, and other growth outcomes and management variables were regressed on TDM milk yield data. In the Cornell herd, preweaning ADG, ranged from 0.10 to 1.58 kg, and was significantly correlated with first-lactation yield; for every 1 kg of preweaning ADG, heifers, on average, produced 850 kg more milk during their first lactation and 235 kg more milk for every Mcal of metabolizable energy intake above maintenance. In the commercial herd, for every 1 kg of preweaning ADG, milk yield increased by 1,113 kg in the first lactation and further, every 1 kg of prepubertal ADG was associated with a 3,281 kg increase in first-lactation milk yield. Among the 2 herds, preweaning ADG accounted for 22% of the variation in first-lactation milk yield as analyzed with the TDM. These results indicate that increased growth rate before weaning results in some form of epigenetic programming that is yet to be understood, but has positive effects on lactation milk yield. This analysis identifies nutrition and management of the preweaned calf as major environmental factors influencing the expression of the genetic capacity of the animal for milk yield.

Technical note: improved methodology for analyses of acid detergent fiber and acid detergent lignin.

The objective of this study was to evaluate the methodology of the acid detergent lignin (ADL) assay in an effort to evaluate particle loss, improve repeatability, and decrease variation within and among samples. The original ADL method relied on asbestos as a filtering aid, but that was removed in 1989 with the mandate from the Environmental Protection Agency to eliminate asbestos in the environment. Furthermore, recent work on fiber methodology indicated that pore size in the Gooch sintered glass crucible (40-60 µm) was too large to trap all of the small particles associated with neutral detergent fiber (NDF) and acid detergent fiber (ADF). Thus, any loss of ADF could potentially result in a loss of ADL. Sixty forages including conventional and brown midrib corn silages, alfalfa silages and hays, mature grasses, early vegetative grasses, and 9 feces samples, were analyzed sequentially for ADF and ADL as outlined in the 1973 procedure of Van Soest except for the use of the asbestos fiber. A glass microfiber filter with a 1.5-µm pore size was chosen as a filtering aid because it met the criteria required by the assay: glass, heat resistant, acid resistant, chemically inert, and hydrophobic. To compare with the current ADF and ADL assays, the assays were conducted with either no filter or the glass filter inserted into crucibles, rinsed with acetone, and then according to the 1973 procedure of Van Soest. The samples analyzed covered a range from 18.11 to 55.79% ADF and from 0.96 to 9.94% ADL on a dry matter (DM) basis. With the use of the filter, the mean ADF values increased 4.2% and mean ADL values increased 18.9%. Overall, both ADF and ADL values were greater with the use of the glass microfiber filter than without, indicating that as the type of sample analyzed changed, use of the Gooch crucible without the filtering aid results in particle loss. The adoption of the use of a small pore size (1.5 µm) glass microfiber filter to improve filtration and recovery of ADF and ADL and to reduce variation in the ADL assay is recommended, especially when sintered glass bottom crucibles are used. These differences in recovery and repeatability have implications for other fiber and lignin methods, as well as for estimating the potential changes in digestibility of fibrous feeds and feed quality.

Effects of Saccharomyces cerevisiae, medium and forage type and their interactions on in vitro ruminal fermentation.

The objective of this study was to investigate the effects of a live yeast, Saccharomyces cerevisiae CNCM I-1077, at four doses (0, 1×105, 1×106 and 1 × 107 cfu/mL) according to the reducing medium used [Goering-Van Soest (GV), McDougall (MD) or Kansas State (KS)] on in vitro ruminal neutral detergent fibre digestibility (NDFd), rate of digestion of NDF (kd), organic matter digestibility (OMd), dry matter digestibility (DMd), pH as well as volatile fatty acids (VFA) concentration, using two forages (oat hay and wheat straw) with differing chemical composition. The maximum in vitro NDFd, DMd, OMd as well as kd were obtained with dose 1 × 106 cfu/mL, although differences between doses were not always significant. The pH estimates were the lowest with the 1 × 107 cfu/mL dose, but the differences were not all significant; however, 1 × 107 cfu/mL corresponded to significantly lower pH estimates compared to the control and 1×105 (6.51 vs. 6.60 and 6.59, respectively). The decrease in pH was accompanied by an increase in VFA concentrations as the yeast dose increased. The KS medium resulted in the lowest digestibility estimates, pH estimates as well as kd, regardless of yeast dose. The 1 × 106 cfu/mL was the better performing yeast dose in vitro resulting in higher digestibility estimates which indicates the yeasts ability to stimulate the microorganisms within the rumen by beneficially modifying rumen environment, thus promoting microbiota activity. The MD and GV media provide better environments for fermentation than the KS medium, resulting in higher in vitro NDFd, DMd, OMd, pH estimates as well as rate of NDF digestion. The MD and GV are also the media that resulted in more consistent results when analysing the effects of the live yeast. Our data suggest that the in vitro conditions have to be carefully chosen to be able to demonstrate rumen fermentation shifts with the use of live microbial additives.

Genotype by environment interaction for yield and somatic cell score with alternative environmental definitions.

Differential genetic expression in high and low opportunity Sicilian Holstein-Friesian and Brown Swiss herd environments was investigated using endogenous and exogenous variables in a set of three definitions. Results of genetic by environmental interaction were compared using alternative environmental definitions: within herd-year standard deviation for mature equivalent milk yield (HYSD), detectable incidence of normal vs. abnormal (peakless) lactation and herds clustered by causal relationships from high and low frequency use of nutrition, milking, health and animal handling practices. Data for genetic analysis consisted of first-lactation standardized yields of milk, fat and protein, and weighted somatic cell score for 8897 daughters of 825 Holstein-Friesian sires and 1143 daughters of 220 Brown Swiss sires. Components of covariance, heritabilities, and genetic correlations were estimated using bivariate and multivariate sire models for average and contrasting environments for each definition. Sire variances for yields were consistently smaller in the low opportunity environments of both breeds. Except for differential incidence of abnormal lactation in Friesian herds, correlated yield response in less privileged environments was 0.41 to 0.81 as much as in high opportunity environments, a substantial loss. Genetic correlations between HYSD environments for yield traits of Friesian were 0.48 to 0.66 but exceeded 0.80 for other definitions. Less correlated response in somatic cell score was also predicted for environments with low use of yield-enhancing practices (0.66 for Friesian and 0.61 for Brown Swiss), which may have resulted from less health care and poorer milking management. Therefore, unfavorable management interactions likely foster unequal gains from selection in contrasting environments defined exogenously or by incidence of peakless lactation. Conversely, greater genetic as well as phenotypic response is expected from additional inputs of nutrition, health care and milking management.