Effects of a phytogenic feed additive on weaned dairy heifer calves subjected to a diurnal heat stress bout.

The study objective was to evaluate the effects of a phytogenic feed additive (PFA) on dry matter intake (DMI), average daily gain (ADG), inflammation, and oxidative stress markers of heifer calves exposed to a heat stress bout in the summer. A total of18 Holstein and 4 Jersey heifer calves (192 ± 5 kg of body weight at 162 ± 16 d of age) housed in indoor stalls were assigned to 1 of 2 dietary treatments (n = 11; 9 Holstein and 2 Jersey): (1) a basal total mixed ration (CTL), and (2) CTL top-dressed with 0.25 g/d of PFA. Following 7 d of acclimation, baseline measurements were made over 7 d under regular summer conditions [average temperature-humidity index (THI) = 79 from 0900 to 2000 h, and 75 from 2000 to 0900 h]. Calves were then subjected to a 7-d cyclic heat stress bout (HS) by turning on barn heaters and increasing the barn temperature to 33.0°C only during the daytime (the average THI = 85 from 0900 to 2000 h). The study continued for an extra 4-d period after HS ended (post-HS). The HS increased rectal temperature, skin temperature, and respiration rate from the baseline by 1.0°C, 4.0°C, and 49 breaths/min, respectively. The drinking water intake increased by 32% in response to HS, and calves continued to consume more water (44%) than the baseline consumption even after HS ended. The treatment × time interactions were not significant for feed intake, ADG, partial pressure of O2 in the blood, and blood concentrations of inflammation markers such as haptoglobin and lipopolysaccharide binding protein (LBP), and antioxidant markers such as protein carbonyl and thiobarbituric acid (TBARS). The PFA tended to increase daytime DMI (0.24 kg/d) compared with CTL throughout the experiment but did not affect ADG, which decreased from 1.12 kg/d to 0.26 kg/d in response to HS. Both DMI (13%) and ADG (85%) increased during post-HS relative to baseline, indicating compensatory performances that were not affected by the PFA. Serum haptoglobin and plasma LBP concentrations of PFA calves were 44% and 38% lower than that of CTL calves across all time points. The PFA decreased O2 pressure and tended to decrease protein carbonyl concentration in the blood across all time points. The PFA tended to decrease TBARS concentration on the first day of HS and increase and decrease the ratio of reduced to oxidized glutathione in the blood during the baseline and post-HS periods, respectively. Despite the lack of growth improvements, feeding PFA seems to increase O2 levels in the blood and alleviate oxidative stress and inflammation of heifer calve exposed to diurnal heat waves (~7 d) in the summer.

Early step-down weaning of dairy calves from a high milk volume with glutamine supplementation.

Weaning dairy calves from a high milk volume (≥8.0 kg/d) can negatively affect the growth and welfare even if it is performed in a step-down manner. Supplementation of Gln improved gut development of preweaning calves and mitigated weaning stresses of piglets to extents achieved with antibiotics. The study objective was to examine the effect of initiating a step-down weaning scheme with a Gln supplement at an early age on calf starter intake (CSI), average daily gain (ADG), and paracellular permeability of the intestinal epithelium of calves fed a high volume of milk (9.0 kg/d). Thirty-six Holstein heifer calves were assigned to 3 treatments (n = 12) as follows: (1) initiating weaning at 49 d of age (LW), (2) initiating weaning at 35 d of age (EW), and (3) initiating weaning at 35 d with a Gln supplement (2.0% of dry matter intake) from 28 to 42 d of age (EWG). Calves were fed 9.0 kg/d of whole milk until weaning was initiated by abruptly decreasing the milk volume to 3.0 kg/d. Weaning was completed once calves achieved ≥1.0 kg/d of CSI. The paracellular permeability of the intestinal epithelium was assessed with lactulose-to-mannitol ratio (LMR) in the blood on 1 d before, and 3 and 7 d after the initiation of weaning. The blood was analyzed for haptoglobin, lipopolysaccharide-binding protein (LBP), and metabolites including AA. The CSI increased once milk volume was restricted in all treatments. The CSI of LW was greater than that of EW and EWG during the first week of weaning. The LW, EW, and EWG took 11, 19, and 16 d to achieve ≥1.0 kg/d of CSI and were weaned at 60, 54, and 51 d of age, respectively. The body weight (BW) of LW, EW, and EWG at the initiation of weaning were 68.2, 58.7, and 59.5 kg, respectively. Both LW and EWG achieved similar ADG, but ADG of EW was lower than LW during the first week of weaning. All calves had similar ADG during the second week of weaning. The BW of LW, EW, and EWG at weaning were 74.8, 66.5, and 66.4 kg, representing a 2.0, 1.8, and 1.8-fold increase in birth weight, respectively. All calves had similar BW of 88.6 and 164.3 kg at 10 and 20 wk of age, respectively. Regardless of the age, serum haptoglobin and plasma LBP concentrations increased on d 3 and returned to baseline concentrations on d 7 during weaning. The EW had a lower plasma LBP concentration than LW and EWG on d 3 during weaning. The LMR was similar between treatments on d 3 but increased by 44% for EW and LW on d 7, whereas the LMR of EWG remained unchanged during weaning. The postprandial serum concentration of Gln, Met, Trp, and ß-hydroxybutyrate were greater for EWG than EW during weaning. Beginning step-down weaning at 35 d with a Gln supplement can help maintain the gut barrier function and wean dairy calves with a satisfactory CSI at 7 wk of age without affecting postweaning growth.

Effects of branched-chain amino acids on glucose uptake and lactose synthesis rates in bovine mammary epithelial cells and lactating mammary tissue slices.

Even though supplementations of essential AA (EAA) are often related to increased lactose yields in dairy cows, underlying mechanisms connecting EAA availability to the mammary glands and lactose synthesis are poorly understood. The objective of this study was to examine the effects of branched-chain AA (BCAA) including Leu, Ile, and Val on (1) glucose transporter (GLUT1) abundance and glucose uptake, (2) the abundance of proteins regulating lactose synthesis pathway, and (3) fractional synthesis rates of lactose (FSR) using bovine mammary epithelial cells (BMEC) and mammary tissues slices (MTS). The BMEC (n = 4) were allocated randomly to regular Dulbecco's Modified Eagle Medium with Ham's F12 (DMEM/F12) medium (+EAA) or +EAA deficient (by 90%) in all EAA (-EAA), all BCAA (-BCAA), only Leu (-Leu), only Ile (-Ile) or only Val (-Val). Western immunoblotting analyses, depletion of glucose in media, and a proteomic analysis were performed to determine the abundance of GLUT1 in the cell membrane, net glucose uptake, and the abundance of enzymes involved in lactose synthesis pathway in BMEC, respectively. The MTS (n = 6) were allocated randomly to DMEM/F12 medium having all EAA and 13C-glucose at concentrations similar to plasma concentrations of cows (+EAAp), and +EAAp deprived of all BCAA (-BCAAp) or only Leu (-Leup) for 3 h. The 13C enrichments of free glucose pool in MTS (EGlu-free) and the enrichments of glucose incorporated into lactose in MTS and media [ELactose-bound (T&M)] were determined and used in calculating FSR. In BMEC, -BCAA increased the fraction of total GLUT1 translocated to the cell membrane and the fraction that was potentially glycosylated compared with +EAA. Among individual BCAA, only -Leu was associated with a 63% increase in GLUT1 translocated to the cell membrane and a 40% increase in glucose uptake of BMEC. The -BCAA tended to be related to a 75% increase in the abundance of hexokinase in BMEC. Deprivation of Leu tended to increase glucose uptake of MTS but did not affect EGlu-free, ELactose-bound (T&M), or FSR relative to +EAAp. On the other hand, -BCAAp did not affect glucose uptake of MTS but was related to lower ELactose-bound (T&M), or FSR relative to +EAAp. Considering together, decreasing Leu supply to mammary tissues enhances GLUT1 and thus glucose uptake, which, however, does not affect lactose synthesis rates. Moreover, the deficiency of other BCAA, Ile, and Val alone or together with the deficiency of Leu seemed to decrease lactose synthesis rates without affecting glucose uptake. The data also emphasize the importance of addressing the effect of the supply of other nutrients to the mammary glands than the precursor supply in describing the synthesis of a milk component.

Energetic metabolism, milk production, and inflammatory response of transition dairy cows fed rumen-protected glucose.

Objectives were to evaluate the effects of rumen-protected glucose (RPG) supplementation on milk production, post-absorptive metabolism, and inflammatory biomarkers in transition dairy cows. Fifty-two multiparous cows were blocked by previous 305-d mature-equivalent milk (305ME) yield and randomly assigned to 1 of 2 iso-energetic and iso-nitrogenous treatments: (1) control diet (CON; n = 26) or (2) a diet containing RPG (pre-fresh 5.3% of dry matter and 6.0% of dry matter postpartum; n = 26). Cows received their respective dietary treatments from d -21 to 28 relative to calving, and dry matter intake was calculated daily during the same period. Weekly body weight, milk composition, and fecal pH were recorded until 28 d in milk (DIM), and milk yield was recorded through 105 DIM. Blood samples were collected on d -7, 3, 7, 14, and 28 relative to calving. Data were analyzed using repeated measures in the MIXED procedure (SAS Institute Inc., Cary, NC) with previous 305ME as a covariate. Fecal pH was similar between treatments and decreased (0.6 units) postpartum. Dry matter intake pre- and postpartum were unaffected by treatment, as was milk yield during the first 28 or 105 DIM. Milk fat, protein, and lactose concentration were similar for both treatments. Blood urea nitrogen and plasma glucose concentrations were unaffected by treatment; however, results showed increased concentration of circulating insulin (27%), lower nonesterified fatty acids (28%), and lower postpartum ß-hydroxybutyrate (24%) in RPG-fed cows. Overall, circulating lipopolysaccharide-binding protein and haptoglobin did not differ by treatment, but at 7 DIM, RPG-fed cows had decreased lipopolysaccharide-binding protein and haptoglobin concentrations (31 and 27%, respectively) compared with controls. Supplemental RPG improved some biomarkers of post-absorptive energetics and inflammation during the periparturient period, changes primarily characterized by increased insulin and decreased nonesterified fatty acids concentrations, with a concomitant reduction in acute phase proteins without changing milk production and composition.

Beginning to offer drinking water at birth increases the species richness and the abundance of Faecalibacterium and Bifidobacterium in the gut of preweaned dairy calves.

We previously demonstrated that dairy calves having access to drinking water since birth (W0) achieved greater body weight, fiber digestibility, and feed efficiency than those that first received drinking water at 17 d of age (W17). Since gut microbiota composition could be linked to growth and development of animals, the objective of this study was to examine the effect of offering drinking water to newborn calves on composition of bacteria in the gut using a fecal microbiota analysis. Fresh feces were collected directly from the rectum of calves in W0 (n = 14) and W17 (n = 15) at 2, 6, and 10 wk of age. All of the calves were fed pasteurized waste milk, weaned at 7 wk of age, and offered tap water according to the treatment. The DNA was sequenced using 16S rRNA gene-amplicon sequencing on an Illumina MiSeq system (Illumina Inc., San Diego, CA). The sequences were clustered into operational taxonomic units (OTU) with a 99% similarity threshold. Treatment effects on α-diversity indices and relative abundance of the 10 most abundant genera were analyzed using GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC). Statistical significance (q-value) of treatment effects on the 50 most abundant OTU was determined with a false discovery rate analysis. At 2 wk of age, W0 had a greater number of observed OTU (5,908 vs. 4,698) and species richness (Chao 1 index) than W17. The number of OTU and richness indices increased from wk 2 to 6, but the increment of W17 was greater than that of W0. The Shannon and inverse-Simpson indices increased linearly with age, but no difference was observed between W0 and W17 at any time point. The Firmicutes to Bacteroidetes ratios were also similar at every time point but decreased markedly when calves were weaned. The relative abundance of genera Faecalibacterium and Bacteroides was greater in W0 than W17 at 2 wk of age. The genus Faecalibacterium continued to be more abundant in W0 than W17 at 6 wk of age but had similar abundance 3 wk after weaning (10 wk of age). The abundance of Faecalibacterium at wk 6 was positively correlated with apparent total-tract digestibility of acid detergent fiber at 10 wk of age. Calves receiving water since birth had greater abundance of OTU related to Faecalibacterium prausnitzii, and Bifidobacterium breve at 6 wk of age (q < 0.085). These species are known to improve growth in preweaned calves. The abundance of none of the genera and OTU was different between W0 at W17 at 10 wk of age (q > 0.100). Overall, beginning to offer drinking water at birth has a potential to modulate gut microbiota composition and thereby positively affect performance of young dairy heifer calves (≤10 wk of age).

Drinking water intake of newborn dairy calves and its effects on feed intake, growth performance, health status, and nutrient digestibility.

Although it is recommended to offer free drinking water (called drinking water hereafter) immediately after birth, producers wait, on average, 17 d to first offer drinking water to newborn dairy calves. The objective of this study was to examine water and feed intake, growth performance, health status, and nutrient digestibility of Holstein heifer calves offered drinking water from birth (W0) as compared with those offered it at 17 d of age (W17), when fed an ad libitum volume of milk. Thirty Holstein heifer calves, balanced for parity of the dam, birth weight, and birth week, were randomly assigned (n = 15) to W0 or W17. Calves had free access to drinking water and a starter ration, offered in 2 separate buckets, until they were 70 d of age. Calves were bottle-fed with pasteurized whole milk 3× per day (2.0 kg/feeding until d 14, and 3.2 kg/feeding thereafter). Calves were partially weaned (33% of the milk allowance 1 × per day) at 42 d of age and completely weaned at 49 d of age. Drinking water intake, starter intake, milk intake, ambient temperature, and the fecal consistency were recorded daily. Body weight, hip height, hip width, heart girth, and body length were measured weekly. Blood (drawn from a jugular vein) was analyzed for hematocrit and haptoglobin concentrations at 14 d of age. On d 69 and 70, total fecal output of individual calves was measured and analyzed for chemical composition to determine apparent total-tract digestibility of nutrients. When offered from birth, newborn calves consumed 0.75 ± 0.05 kg/d water aside from the water they received from ad libitum milk allowance during the first 16 d. Once offered, W17 calves drank more water (59%) than W0 calves during the preweaning period. Starter intake of W0 and W17 calves was similar, but W0 calves consumed 0.285 kg/d more milk and tended to achieve greater body weight and heart girth compared with W17 calves during the preweaning period. Offering water from birth versus offering it later did not affect the number of days with diarrhea, intensity of diarrhea, or blood hematocrit and haptoglobin concentrations of preweaned calves. Despite a similar starter intake, W0 calves had greater hip height, body length, apparent total-tract digestibility of acid detergent fiber and neutral detergent fiber, and feed efficiency than W17 calves postweaning (50 to 70 d of age). When followed up to 5 mo of age, W0 calves had greater body weight than W17 calves. Provision of drinking water immediately after birth could improve growth and development of calves pre- and postweaning, potentially by stimulating rumen development, thus increasing nutrient availability.

Effect of Dairy Manure Storage Conditions on the Survival of E. coli O157:H7 and Listeria.

Dairy manure is regularly applied to crop fields as a solid or liquid to improve the soil nutrient status. However, pathogens may survive during manure storage and enter the environment during application. In this study, three storage practices were evaluated to understand the survival patterns of O157:H7 and spp. in dairy manure using a culture-based approach. To replicate common farm manure storage techniques, solid manure was stacked as piles with periodic turning or as static piles without turning, whereas liquid manure (feces, urine, and water) was stored as a slurry in small tanks to simulate lagoon conditions. The and levels in the manure samples were determined for 29 wk. Results showed that there was an initial reduction in bacteria levels in the first month; however, both and managed to survive in the solid manure piles for the full study period. In slurry samples, was not detected after 14 wk, but survived until the end of the experiment at relatively lower levels than in the solid manure piles. Ambient weather and pile size were identified as the main reasons for bacteria survival during the course of the experiment. The outcome of this study is important in terms of understanding pathogen survival in manure piles and slurries prior to their application to crop fields.

Predicting manure volatile solid output of lactating dairy cows.

Organic matter (OM) in livestock manure consisting of biodegradable and nonbiodegradable fractions is known as volatile solids (VS). According to the Intergovernmental Panel on Climate Change (IPCC) Tier 2 guidelines, methane produced by stored manure is determined based on VS. However, only biodegradable OM generates methane production. Therefore, estimates of biodegradable VS (dVS; dVS = VS - lignin) would yield better estimates of methane emissions from manure. The objective of the study was to develop mathematical models for estimating VS and dVS outputs of lactating dairy cows. Dry matter intake, dietary nutrient contents, milk yield and composition, body weight, and days in milk were used as potential predictor variables. Multicollinearity, model simplicity, and random study effects were taken into account during model development that used 857 VS and dVS measurements made on individual cows (kg/cow per day) from 43 metabolic trials conducted at the USDA Energy and Metabolism laboratory in Beltsville, Maryland. The new models and the IPCC Tier 2 model were evaluated with an independent data set including 209 VS and dVS measurements (kg/cow per day) from 2 metabolic trials conducted at the University of California, Davis. Organic matter intake (kg/d) and dietary crude protein and neutral detergent fiber contents (% of dry matter) were significantly associated with VS. A new model including these variables fitted best to data. When evaluated with independent data, the new model had a root mean squared prediction error as a percentage of average observed value (RMSPE) of 12.5%. Mean and slope biases were negligible at <1% of total prediction bias. When energy digestibility of the diet was assumed to be 67%, the IPCC Tier 2 model had a RMSPE of 13.7% and a notable mean bias for VS to be overpredicted by 0.4 kg/cow per day. A separate model including OM intake as well as dietary crude protein and neutral detergent fiber contents as predictor variables fitted best to dVS data and performed well on independent data (RMSPE = 12.7%). The Cornell Net Carbohydrate and Protein System model relying on fat-corrected milk yield and body weight more successfully predicted dry matter intake (DMI; RMSPE = 14.1%) than the simplified (RMSPE = 16.9%) and comprehensive (RMSPE = 23.4%) models to predict DMI in IPCC Tier 2 methodology. New models and the IPCC Tier 2 model using DMI from the Cornell Net Carbohydrate and Protein System model predicted VS (RMSPE = 17.7-19.4%) and dVS (RMSPE = 20%) well with small systematic bias (<10% of total bias). The present study offers empirical models that can accurately predict VS and dVS of dairy cows using routinely available data in dairy farms and thereby assist in efficiently determining methane emissions from stored manure.

Exogenous ß-mannanase improves feed conversion efficiency and reduces somatic cell count in dairy cattle.

Exogenous fibrolytic enzymes have been shown to be a promising way to improve feed conversion efficiency (FCE). ß-Mannanase is an important enzyme digesting the polysaccharide ß-mannan in hemicellulose. Supplementation of diets with ß-mannanase to improve FCE has been more extensively studied in nonruminants than in ruminants. The objective of this study was to investigate the effects of ß-mannanase supplementation on nutrient digestibility, FCE, and nitrogen utilization in lactating Holstein dairy cows. Twelve post-peak-lactation multiparous Holstein cows producing 45.5±6.6kg/d of milk at 116±19.0d in milk were randomly allotted to 1 of 3 treatments in a 3×3 Latin square design with 3 periods of 18d (15d for adaptation plus 3d for sample collection). All cows were fed the same basal diet and the 3 treatments differed only by the ß-mannanase dose: 0% dry matter (DM; control), 0.1% of DM (low supplement, LS), and 0.2% of DM (high supplement, HS) supplemented to the basal diet. Supplementation of ß-mannanase enzyme at the LS dose reduced dry matter intake (DMI) but did not affect milk yield or milk composition. Cows receiving LS produced 90g more milk per kg of DMI compared with control cows. Somatic cell count (SCC) in milk was lower for cows fed the LS diet compared with cows fed control diets. Cows fed LS diet had lower DM, organic matter and crude protein digestibility compared with cows fed control diets. Starch, neutral detergent fiber, and acid detergent fiber digestibility were not affected by LS. Milk yield, DMI, SCC, and nutrient digestibility did not change for HS. Despite the reduced crude protein digestibility, reduced N intake led to similar fecal N excretions in LS cows and control cows (234 vs. 235g/cow per day). Urinary N excretions remained similar between enzyme-fed and control cows (~190g/cow per day), although the percentage of N intake partitioned to urinary N tended to be greater in LS than in control cows (31 vs. 27%). Cows fed LS significantly improved the percentage of apparently absorbed N partitioned to milk protein N (42 vs. 38%). When supplemented at 0.1% of dietary DM, ß-mannanase can improve FCE and lower the SCC of dairy cows without affecting milk yield, milk composition, or total manure N excretions of dairy cows.

Prediction of drinking water intake by dairy cows.

Mathematical models that predict water intake by drinking, also known as free water intake (FWI), are useful in understanding water supply needed by animals on dairy farms. The majority of extant mathematical models for predicting FWI of dairy cows have been developed with data sets representing similar experimental conditions, not evaluated with modern cows, and often require dry matter intake (DMI) data, which may not be routinely available. The objectives of the study were to (1) develop a set of new empirical models for predicting FWI of lactating and dry cows with and without DMI using literature data, and (2) evaluate the new and the extant models using an independent set of FWI measurements made on modern cows. Random effect meta-regression analyses were conducted using 72 and 188 FWI treatment means with and without dietary electrolyte and daily mean ambient temperature (TMP) records, respectively, for lactating cows, and 19 FWI treatment means for dry cows. Milk yield, DMI, body weight, days in milk, dietary macro-nutrient contents, an aggregate milliequivalent concentration of dietary sodium and potassium (NaK), and TMP were used as potential covariates to the models. A model having positive relationships of DMI, dietary dry matter (DM%), and CP (CP%) contents, NaK, and TMP explained 76% of variability in FWI treatment means of lactating cows. When challenged on an independent data set (n=261), the model more accurately predicted FWI [root mean square prediction error as a percentage of average observed value (RMSPE%)=14.4%] compared with a model developed without NaK and TMP (RMSPE%=17.3%), and all extant models (RMSPE%≥15.7%). A model without DMI included positive relationships of milk yield, DM%, NaK, TMP, and days in milk, and explained 63% of variability in the FWI treatment means and performed well (RMSPE%=17.9%), when challenged on the independent data. New models for dry cows included positive relationships of DM% and TMP along with DMI or body weight. The new models with and without DMI explained 75 and 54% of the variability in FWI treatment means of dry cows and had RMSPE% of 12.8 and 15.2%, respectively, when evaluated with the literature data. The study offers a set of empirical models that can assist in determining drinking water needs of dairy farms.

Development of a model describing regulation of casein synthesis by the mammalian target of rapamycin (mTOR) signaling pathway in response to insulin, amino acids, and acetate.

To improve dietary protein use efficiency in lactating cows, mammary protein synthesis responses to AA, energy substrates, and hormones must be better understood. These entities exert their effects through stimulation of mRNA translation via control of initiation and elongation rates at the cellular level. A central protein kinase of this phenomenon is the mammalian target of rapamycin (mTOR), which transfers the nutritional and hormonal stimuli onto a series of proteins downstream through a cascade of phosphorylation reactions that ultimately affect protein synthesis. The objective of this work was to further develop an existing mechanistic model of mTOR phosphorylation responses to insulin and total essential AA to include the effects of specific essential AA and acetate mediated by signaling proteins including protein kinase B (Akt), adenosine monophosphate activated protein kinase (AMPK), and mTOR and to add a representation of milk protein synthesis. Data from 6 experiments in MAC-T cells and mammary tissue slices previously conducted in our laboratory were assembled and used to parameterize the dynamic system of differential equations representing Akt, AMPK, and mTOR in their phosphorylated and dephosphorylated states and the resulting regulation of milk protein synthesis. The model predicted phosphorylated Akt, mTOR, AMPK, and casein synthesis rates with root mean square prediction errors of 16.8, 28.4, 33.0, and 54.9%, respectively. All other dependent variables were free of mean and slope bias, indicating an adequate representation of the data. Whereas mTOR was not very sensitive to changes in insulin or acetate levels, it was highly sensitive to leucine and isoleucine, and this signal appeared to be effectively transduced to casein synthesis. Although prior work had observed a relationship with additional essential AA, and data supporting those conclusions were present in the data set, we were unable to derive significant relationships with any essential AA other than leucine and isoleucine. The signaling properties and dynamics of AMPK under nutrient depletion and sufficiency, the responses to additional essential AA, and the consequent effects on protein synthesis remain to be better understood.

Prediction of phosphorus output in manure and milk by lactating dairy cows.

Mathematical models for predicting P excretions play a key role in evaluating P use efficiency and monitoring the environmental impact of dairy cows. However, the majority of extant models require feed intake as predictor variable, which is not routinely available at farm level. The objectives of the study were to (1) explore factors explaining heterogeneity in P output; (2) develop a set of empirical models for predicting P output in feces (Pf), manure (PMa), and milk (Pm, all in g/cow per day) with and without dry matter intake (DMI) using literature data; and (3) evaluate new and extant P models using an independent data set. Random effect meta-regression analyses were conducted using 190 Pf, 97 PMa, and 118 Pm or milk P concentration (PMilkC) treatment means from 38 studies. Dietary nutrient composition, milk yield and composition, and days in milk were used as potential covariates to the models with and without DMI. Dietary phosphorus intake (Pi) was the major determinant of Pf and PMa. Milk yield negatively affected Pi partitioning to Pf or PMa. In the absence of DMI, milk yield, body weight, and dietary P content became the major determinants of Pf and PMa. Milk P concentration (PMilkC) was heterogeneous across the treatment groups, with a mean of 0.92 g/kg of milk. Milk yield, days in milk, and dietary Ca-to-ash ratio were negatively correlated with PMilkC and explained 42% of the heterogeneity. The new models predicted Pf and PMa with root mean square prediction error as a percentage of observed mean (RMSPE%) of 18.3 and 19.2%, respectively, using DMI when evaluated with an independent data set. Some of the extant models also predicted Pf and PMa well (RMSPE%=19.3 to 20.0%) using DMI. The new models without DMI as a variable predicted Pf and PMa with RMSPE% of 22.3 and 19.6%, respectively, which can be used in monitoring P excretions at farm level. When evaluated with an independent data set, the new model and extant models based on milk protein content predicted PMilkC with RMSPE% of 12.7 to 19.6%. Although models using P intake information gave better predictions, P output from lactating dairy cows can also be predicted well without intake using milk yield, milk protein content, body weight, and dietary P, Ca, and total ash contents.

Quantifying body water kinetics and fecal and urinary water output from lactating Holstein dairy cows.

Reliable estimates of fresh manure water output from dairy cows help to improve storage design, enhance efficiency of land application, quantify the water footprint, and predict nutrient transformations during manure storage. The objective of the study was to construct a mechanistic, dynamic, and deterministic mathematical model to quantify urinary and fecal water outputs (kg/d) from individual lactating dairy cows. The model contained 4 body water pools: reticulorumen (QRR), post-reticulorumen (QPR), extracellular (QEC), and intracellular (QIC). Dry matter (DM) intake, dietary forage, DM, crude protein, acid detergent fiber and ash contents, milk yield, and milk fat and protein contents, days in milk, and body weight were input variables to the model. A set of linear equations was constructed to determine drinking, feed, and saliva water inputs to QRR and fractional water passage from QRR to QPR. Water transfer via the rumen wall was subjected to changes in QEC and total water input to QRR. Post-reticulorumen water passage was adjusted for DM intake. Metabolic water production and respiratory cutaneous water losses were estimated with functions of heat production in the model. Water loss in urine was driven by absorbed N left after being removed via milk. Model parameters were estimated simultaneously using observed fecal and urinary water output data from lactating Holstein cows (n=670). The model was evaluated with data that were not used for model development and optimization (n=377). The observations in both data sets were related to thermoneutral conditions. The model predicted drinking water intake, fecal, urinary, and total fresh manure water output with root mean square prediction errors as a percentage of average values of 18.1, 15.6, 30.6, and 14.6%, respectively. In all cases, >97% of the prediction error was due to random variability of data. The model can also be used to determine saliva production, heat and metabolic water production, respiratory cutaneous water losses, and size of major body water pools in lactating Holstein cows under thermoneutral conditions.

Effects of phytase supplementation on phosphorus retention in broilers and layers: a meta-analysis.

Phytase, a widely used feed additive in poultry diets, increases P availability and subsequently reduces inorganic-P supplementation and P-excretion. Phytase supplementation effect on P-retention in poultry has been investigated, but the effect sizes were highly variable. The present study's objective was to conduct several meta-analyses to quantitatively summarize the phytase effect on P-retention in broilers and layers. Data from 103 and 26 controlled experiments testing the phytase effect on P-retention were included in 2 separate meta-analyses for broilers and layers, respectively. The mean difference calculated by subtracting the means of P-retention for the control group from the phytase-supplemented group was chosen as an effect size estimate. Between-study variability (heterogeneity) of mean difference was estimated using random-effect models and had a significant effect (P < 0.01) in both broilers and layers. Therefore, random-effect models were extended to mixed-effect models to explain heterogeneity and obtain final phytase effect size estimates. Available dietary and bird variables were included as fixed effects in the mixed-effect models. The final broiler mixed-effect model included phytase dose and Ca-to-total-P ratio (Ca:tP), explaining 15.6% of the heterogeneity. Other variables such as breed might further explain between-study variance. Broilers consuming control diets were associated with 48.4% P-retention. Exogenous phytase supplementation at 1,039 FTU/kg of diet increased P-retention by 8.6 percentage units on average. A unit increase of phytase dose and Ca:tP from their means further increased P-retention. For layers, the final mixed-effect models included dietary Ca, age, and experimental period length. The variables explained 65.9% of the heterogeneity. Layers receiving exogenous phytase at 371 FTU/kg were associated with a 5.02 percentage unit increase in P-retention. A unit increase in dietary Ca from its mean increased P-retention, whereas an increase in the experiment length and layer's age decreased P-retention. Phytase supplementation had a significant positive effect on P-retention in both broilers and layers, but effect sizes across studies were significantly heterogeneous due to differences in Ca contents, experiment length, bird age, and phytase dose.

Effects of AMP-activated protein kinase (AMPK) signaling and essential amino acids on mammalian target of rapamycin (mTOR) signaling and protein synthesis rates in mammary cells.

Regulation of mammary protein synthesis potentially changes the relationships between AA supply and milk protein output represented in current nutrient requirement models. Glucose and AA regulate muscle protein synthesis via cellular signaling pathways involving mammalian target of rapamycin (mTOR) and AMP-activated protein kinase (AMPK). The objective of this study was to investigate the effects of essential AA (EAA) and acetate or glucose on mTOR and AMPK signaling pathways and milk protein synthesis rates. A bovine mammary epithelial cell line, MAC-T, was subjected to different media containing 0 or 3.5 mmol/L EAA concentrations with 0 or 5 mmol/L acetate or 0 or 17.5 mmol/L glucose in 2 separate 2 × 2 factorial studies. In a separate set of experiments, lactogenic bovine mammary tissue slices were subjected to the same treatments except that the low EAA treatment contained a low level of EAA (0.18 mmol/L). Supplementation of EAA enhanced phosphorylation of mTOR (Ser2448) and eukaryotic initiation factor 4E binding protein 1 (4EBP1, Thr37/46), and reduced phosphorylation of eukaryotic elongation factor 2 (eEF2, Thr56) in MAC-T cells. Concentration of ATP and phosphorylation of AMPK increased and decreased, respectively, in the presence of EAA in MAC-T cells. Acetate, EAA, or glucose numerically reduced AMPK phosphorylation by about 16% in mammary tissue slices. Provision of EAA increased phosphorylation of mTOR and 4EBP1, intracellular total EAA concentration, and casein synthesis rates in mammary tissue slices, irrespective of the presence of acetate or glucose in the medium. Phosphorylation of mTOR had a marginally negative association with AMPK phosphorylation, which was positively related to eEF2 phosphorylation. Casein synthesis rates were positively and more strongly linked to mTOR phosphorylation than the negative link between eEF2 phosphorylation and casein synthesis rates. A 100% increase in mTOR phosphorylation was associated with an increase in the casein synthesis rate of 0.74%·h(-1), whereas a 100% increase in eEF2 phosphorylation was related to a decline in the casein synthesis rate of 0.33%·h(-1). Although AMPK phosphorylation was responsive to cellular energy status and had a negative effect on mTOR-mediated signals in bovine mammary epithelial cells, its effect on milk protein synthesis rates appeared to be marginal compared with the mTOR-mediated regulation of milk protein synthesis by EAA.

Revised digestive parameter estimates for the Molly cow model.

The Molly cow model represents nutrient digestion and metabolism based on a mechanistic representation of the key biological elements. Digestive parameters were derived ad hoc from literature observations or were assumed. Preliminary work determined that several of these parameters did not represent the true relationships. The current work was undertaken to derive ruminal and postruminal digestive parameters and to use a meta-approach to assess the effects of interactions among nutrients and identify areas of model weakness. Model predictions were compared with a database of literature observations containing 233 treatment means. Mean square prediction errors were assessed to characterize model performance. Ruminal pH prediction equations had substantial mean bias, which caused problems in fiber digestion and microbial growth predictions. The pH prediction equation was reparameterized simultaneously with the several ruminal and postruminal digestion parameters, resulting in more realistic parameter estimates for ruminal fiber digestion, and moderate reductions in prediction errors for pH, neutral detergent fiber, acid detergent fiber, and microbial N outflow from the rumen; and postruminal digestion of neutral detergent fiber, acid detergent fiber, and protein. Prediction errors are still large for ruminal ammonia and outflow of starch from the rumen. The gain in microbial efficiency associated with fat feeding was found to be more than twice the original estimate, but in contrast to prior assumptions, fat feeding did not exert negative effects on fiber and protein degradation in the rumen. Microbial responses to ruminal ammonia concentrations were half saturated at 0.2mM versus the original estimate of 1.2mM. Residuals analyses indicated that additional progress could be made in predicting microbial N outflow, volatile fatty acid production and concentrations, and cycling of N between blood and the rumen. These additional corrections should lead to an even more robust representation of the effects of dietary nutrients on ruminal metabolism and nutrient absorption, of animal performance, and the environmental impact of dairy production.

Effects of jugular-infused lysine, methionine, and branched-chain amino acids on milk protein synthesis in high-producing dairy cows.

In addition to lysine and methionine, current ration-balancing programs suggest that branched-chain amino acid (BCAA) supply may also be limiting in dairy cows. The objective of this study was to investigate whether BCAA, leucine, isoleucine, and valine become limiting for milk protein synthesis when methionine and lysine supply were not limiting. Nine multiparous Holstein cows with an average milk production of 53.5±7.1 kg/d were randomly assigned to 7-d continuous jugular infusions of saline (CTL), methionine and lysine (ML; 12 g and 21 g/d, respectively), or ML plus leucine, isoleucine, and valine (ML+BCAA; 35 g, 15 g, and 15 g/d, respectively) in a 3×3 Latin square design with 3 infusion periods separated by 7-d noninfusion periods. The basal diet consisted of 40% corn silage, 14% alfalfa hay, and a concentrate mix, and respectively supplied lysine, methionine, isoleucine, leucine, and valine as 6.1, 1.8, 4.7, 8.9, and 5.3% of metabolizable protein. Dry matter intake (23.9 kg/d), milk yield (52.8 kg/d), fat content (2.55%), fat yield (1.33 kg/d), lactose content (4.77%), lactose yield (2.51 kg/d), and milk protein efficiency (0.38) were similar across treatments. Protein yield and protein content were not significantly different between ML (1.52 kg/d and 2.88%, respectively) and ML+BCAA (1.51 kg/d and 2.83%, respectively), but they were significantly greater than that of CTL (1.39 kg/d and 2.71%). Cows that received ML+BCAA had less milk urea nitrogen content (10.9 mg/dL) compared with milk of CTL cows (12.4 mg/dL) and ML cows (11.8 mg/dL). Whereas high-producing cows responded positively to methionine and lysine supplementation, no apparent benefits of BCAA supplementation in milk protein synthesis were found. Infusion of BCAA may have stimulated synthesis of other body proteins, probably muscle proteins, as evidenced by decreased milk urea nitrogen.

Regulation of protein synthesis in mammary glands of lactating dairy cows by starch and amino acids.

The objective of this study was to evaluate local molecular adaptations proposed to regulate protein synthesis in the mammary glands. It was hypothesized that AA and energy-yielding substrates independently regulate AA metabolism and protein synthesis in mammary glands by a combination of systemic and local mechanisms. Six primiparous mid-lactation Holstein cows with ruminal cannulas were randomly assigned to 4 treatment sequences in a replicated incomplete 4 x 4 Latin square design experiment. Treatments were abomasal infusions of casein and starch in a 2 x 2 factorial arrangement. All animals received the same basal diet (17.6% crude protein and 6.61 MJ of net energy for lactation/kg of DM) throughout the study. Cows were restricted to 70% of ad libitum intake and abomasally infused for 36 h with water, casein (0.86 kg/d), starch (2 kg/d), or a combination (2 kg/d starch+0.86 kg/d casein) using peristaltic pumps. Milk yields and composition were assessed throughout the study. Arterial and venous plasma samples were collected every 20 min during the last 8h of infusion to assess mammary uptake. Mammary biopsy samples were collected at the end of each infusion and assessed for the phosphorylation state of selected intracellular signaling molecules that regulate protein synthesis. Animals infused with casein had increased arterial concentrations of AA, increased mammary extraction of AA from plasma, either no change or a trend for reduced mammary AA clearance rates, and no change in milk protein yield. Animals infused with starch had increased milk and milk protein yields, increased mammary plasma flow, reduced arterial concentrations of AA, and increased mammary clearance rates and net uptake of some AA. Infusions of starch increased plasma concentrations of glucose, insulin, and insulin-like growth factor-I. Starch infusions increased phosphorylation of ribosomal protein S6 and endothelial nitric oxide synthase, consistent with changes in milk protein yields and plasma flow, respectively. Phosphorylation of the mammalian target of rapamycin was increased in response to starch only when casein was also infused. Thus, cell signaling molecules involved in the regulation of protein synthesis differentially responded to these nutritional stimuli. The hypothesized independent effects of casein and starch on animal metabolism and cell signaling were not observed, presumably because of the lack of a milk protein response to infused casein.

Phenotypic and genetic relationships of common health disorders with milk and fat yield persistencies from producer-recorded health data and test-day yields.

The objective of this study was to investigate phenotypic and genetic relationships of common health disorders in dairy cows with milk (PMY) and fat (PFY) yield persistencies. Health and production data from 398 commercial dairy herds were used. Disease traits were defined in binary form for individual lactations considering mastitis only during the first 100 d in milk (MAST1), only after 100 d in milk (MAST2), and at any stage of lactation (MAST), and reproductive disorders (REPRO), metabolic disorders (METAB), and lameness (LAME). The persistencies were defined to be uncorrelated with 305-d yields. Impact of the diseases on PMY and PFY were investigated separately in first (FL) and later (LL) lactations. Phenotypic associations of PMY and PFY with likelihood of diseases in current and subsequent lactations were examined using odds ratios from a logistic regression model. Linear-threshold sire-maternal grandsire models were used to estimate genetic correlations of displaced abomasums (DA), ketosis (KET), metritis (MET), MAST, MAST1, and MAST2 with PMY and PFY across parities. Metabolic diseases and REPRO had significantly positive relationships with PMY and PFY in both FL and LL. Significantly greater PMY and PFY were associated with MAST1 in LL. Significantly lower PMY and PFY were related to MAST2 in both FL and LL, whereas cows affected by MAST had significantly less persistent lactations. Incidence of MAST and MAST2 decreased with increasing PMY and PFY in the present and previous lactation. Heritability of disease incidences were 0.03 (DA), 0.01 (KET), 0.10 (MAST), 0.02 to 0.05 (MAST1), 0.02 (MAST2), and 0.04 to 0.10 (MET). Displaced abomasum, KET, MAST, MAST1, and MET had unfavorable genetic correlations of 0.35, 0.46, 0.17, 0.02, and 0.27 with PMY, and 0.16, 0.21, 0.07, 0.06, and 0.12 with PFY, respectively. Favorable genetic correlations were found for MAST2 with PMY (-0.24) and PFY (-0.04). Results suggest that diseases in early lactation increase persistency of milk and fat yield. Selection for greater lactation persistency must consider these antagonistic relationships.

Heritability of electronically recorded daily body weight and correlations with yield, dry matter intake, and body condition score.

The objectives of this study were to estimate heritability for daily body weight (BW) and genetic correlations of daily BW with daily milk yield (MY), body condition score (BCS), dry matter intake, fat yield, and protein yield. The Afiweigh cow body weighing system records BW of every cow exiting the milking parlor. The Afiweigh system was installed at the Pennsylvania State University dairy herd in August 2001 and in July 2004 at the Virginia Tech dairy herd. The edited data included 202,143 daily BW and 290,930 daily MY observations from 575 Pennsylvania State University and 120 Virginia Tech Holstein cows. Data were initially analyzed with a series of 4-trait animal models, followed by random regression models. The models included fixed effects for age within lactation group, week of lactation, and herd-date. Random effects included animal, permanent environment, and error. The order of the polynomials for random animal and permanent environmental effects with the random regression model for daily BW was 4 and 6, respectively. Heritability estimates for daily BW ranged from 0.48 to 0.57 and were largest between 200 and 230 and smallest at 305 d of lactation. Genetic correlations were large between BW and BCS (0.60). The genetic correlation between daily BW and MY was -0.14 but was positive (0.24) after adjusting for BCS. Electronically recorded daily BW is highly heritable, and genetic evaluations of daily BW and BW change across the lactation could be used to select for less early lactation BW loss.