Milk beverage base with lactose removed with ultrafiltration: Effect of fat and protein concentration on sensory and physical properties.

Our objectives were to determine the effect of fat (skim to whole milk) and protein (3.4%-10.5%) concentration on the sensory and physical properties of milk beverage base that had lactose and other low molecular components removed by ultrafiltration (UF). In experiment 1, a matrix of 16 treatments was produced to achieve 4 levels of lactose removal (0%, 30%, 70%, and 97%) at each of 4 fat levels (skim, 1%, 2%, and whole milk). In experiment 2, a matrix of 12 treatments was produced to achieve 4 levels of lactose removal (0%, 30%, 70%, and 97%) at each of 3 protein concentrations (3.4%, 6.5%, and 10.5% protein). Physical and sensory properties of these products were determined. Removal of >95% of milk lactose by UF required a diafiltration volume of approximately 3 times the milk volume. Lactose and low molecular weight solute removal increased whiteness across the range from skim to whole milk while decreasing viscosity and making milk flavor blander. In addition, lactose (and other low molecular weight solute) removal by UF decreased titratable acidity by more than 50% and increased milk pH at 20°C to >7.0. Future work on milk and milk-based beverages with lactose removed by UF needs to focus on interaction of the remaining milk solids with added flavorings, changing casein to whey protein ratio before removal of lactose by UF, and the effect of lactose and low molecular weight solute removal on heat stability, particularly for neutral-pH, shelf-stable milk-based beverages.

Effect of replacing a portion of inorganic chloride trace minerals with trace mineral amino acid complexes.

The objective was to determine whether replacing a portion of inorganic chloride trace minerals and cobalt carbonate in the diet with AA complexes of trace minerals and cobalt glucoheptonate will improve lactating cow performance, feed efficiency, and calf performance. In a clinical trial, 69 Holstein cows entering second lactation and greater were randomly assigned to 1 of 2 treatments, with the total dietary trace mineral concentration the same between treatments, starting 1 wk after dry off (50 to 57 d before expected parturition) until 154 d in milk (DIM): (1) an inorganic chloride trace mineral (ITM) blend consisting of Zn (75 mg/kg), Mn (65 mg/kg), and Cu (10 mg/kg) as hydroxychlorides and Co (1 mg/kg) as carbonate (n = 37) or (2) partial replacement of ITM with AA complexes of Zn (40 mg/kg), Mn (20 mg/kg), and Cu (3.5 mg/kg) and Co glucoheptonate (1 mg/kg; AATM; Availa-Dairy, Zinpro Corp.; n = 32). Dry matter intake (DMI) was recorded daily from enrollment through wk 8, and milk yields were recorded daily from calving through wk 22. Milk composition and body weights (BW) were collected weekly. Serum samples were analyzed for albumin (Alb), cholesterol (Chol), total bilirubin (Bili), aspartate aminotransferase (AST), haptoglobin, ß-hydroxybutyrate (BHB), and Ca. A liver health index (LHI) was calculated based on Bili, Chol, and Alb concentrations. A liver functionality index (LFI) was calculated to standardize changes in Alb, Chol, and Bili from 4 to 29 DIM. Greater LHI and LFI indicate better health status. Colostrum was analyzed for IgG and Brix, and calf serum was analyzed for IgG. Calf growth was monitored through 9 wk of age (AATM: n = 12, ITM: n = 10). Data were analyzed using SAS software with mixed effects models and repeated-measures analysis, when applicable. Survival analysis for pregnancy by 154 DIM was analyzed by Cox proportional and Kaplan-Meier hazards models. Disorder incidence was tested with Fisher's exact test. Prepartum DMI as a percent of BW was lower in cows fed AATM and not significant postpartum. Cows fed AATM produced more milk from wk 1 to 8 and from wk 1 to 22. Energy-corrected milk yield and colostrum measures did not significantly differ between treatments. A treatment by time interaction was seen for AST and BHB; cows fed AATM tended to have lower AST concentrations at 28 DIM and lower concentrations in BHB through 29 DIM, though not statistically significant. Cows fed AATM had greater LHI at 4 DIM. Haptoglobin, Ca, LFI, hazard of pregnancy, risk to first service, survival curves, or services per pregnancy did not significantly differ. Calf serum IgG and birth weight did not significantly differ between treatments. Calves from dams fed AATM had greater average daily gain than calves from dams fed ITM. Overall, cows fed AATM during the dry period and early lactation had improved postpartum performance and potential health improvements.

Calcium dynamics and associated temporal patterns of milk constituents in early-lactation multiparous Holsteins.

At the onset of lactation, calcium (Ca) homeostasis is challenged. For the transitioning dairy cow, inadequate responses to this challenge may result in subclinical hypocalcemia at some point in the postpartum period. It has been proposed that dynamics of blood Ca and the timing of subclinical hypocalcemia allow cows to be classified into 4 Ca dynamic groups by assessing serum total Ca concentrations (tCa) at 1 and 4 days in milk (DIM). These differing dynamics are associated with different risks of adverse health events and suboptimal production. Our prospective cohort study aimed to characterize the temporal patterns of milk constituents in cows with differing Ca dynamics to investigate the potential of Fourier-transform infrared spectroscopic (FTIR) analysis of milk as a diagnostic tool for identifying cows with unfavorable Ca dynamics. We sampled the blood of 343 multiparous Holsteins on a single dairy in Cayuga County, New York, at 1 and 4 DIM and classified these cows into Ca dynamic groups using threshold concentrations of tCa (1 DIM: tCa <1.98 mmol/L; 4 DIM: tCa <2.22 mmol/L) derived from receiver operating characteristic curve analysis based on epidemiologically relevant health and production outcomes. We also collected proportional milk samples from each of these cows from 3 to 10 DIM for FTIR analysis of milk constituents. Through this analysis we estimated the milk constituent levels of anhydrous lactose (g/100 g of milk and g/milking), true protein (g/100 g of milk and g/milking), fat (g/100 g of milk and g/milking), milk urea nitrogen (mg/100 g of milk), fatty acid (FA) groups including de novo, mixed origin, and preformed FA measured in grams/100 g of milk, by relative percentage, and grams/milking, as well as energy-related metabolites including ketone bodies and milk-predicted blood nonesterified FA. Individual milk constituents were compared among groups at each time point and over the entire sample period using linear regression models. Overall, we found differences among the constituent profiles of Ca dynamic groups at approximately every time point and over the entire sample period. The 2 at-risk groups of cows did not differ from each other at more than one time point for any constituent, however prominent differences existed between the milk of normocalcemic cows and the milk of the other Ca dynamic groups with respect to FA. Over the entire sample period, lactose and protein yield (g/milking) were lower in the milk of at-risk cows than in the milk of the other Ca dynamic groups. In addition, milk yield per milking followed patterns consistent with previous Ca dynamic group research. Though our use of a single farm does limit the general applicability of these findings, our conclusions provide evidence that FTIR may be a useful method for discriminating between cows with different Ca dynamics at time points that may be relevant in the optimization of management or development of clinical intervention strategies.

Patterns of Fourier-transform infrared estimated milk constituents in early lactation Holstein cows on a single New York State dairy.

Cows undergo immense physiological stress to produce milk during early lactation. Monitoring early lactation milk through Fourier-transform infrared (FTIR) spectroscopy might offer an understanding of which cows transition successfully. Daily patterns of milk constituents in early lactation have yet to be reported continuously, and the study objective was to initially describe these patterns for cows of varying parity groups from 3 through 10 d postpartum, piloted on a single dairy. We enrolled 1,024 Holstein cows from a commercial dairy farm in Cayuga County, New York, in an observational study, with a total of 306 parity 1 cows, 274 parity 2 cows, and 444 parity ≥3 cows. Cows were sampled once daily, Monday through Friday, via proportional milk samplers, and milk was stored at 4°C until analysis using FTIR. Estimated constituents included anhydrous lactose, true protein, and fat (g/100 g of milk); relative % (rel%) of total fatty acids (FA) and concentration (g/100 g of milk) of de novo, mixed, and preformed FA; individual fatty acids C16:0, C18:0, and C18:1 cis-9 (g/100 g of milk); milk urea nitrogen (MUN; mg/100 g of milk); and milk acetone (mACE), milk ß-hydroxybutyrate (mBHB), and milk-predicted blood nonesterified fatty acids (mpbNEFA) (all expressed in mmol/L). Differences between parity groups were assessed using repeated-measures ANOVA. Milk yield per milking differed over time between 3 and 10 DIM and averaged 8.7, 13.3, and 13.3 kg for parity 1, 2, and ≥3 cows, respectively. Parity differences were found for % anhydrous lactose, % fat, and preformed FA (g/100 g of milk). Parity differed across DIM for % true protein, de novo FA (rel% and g/100 g of milk), mixed FA (rel% and g/100 g of milk), preformed FA rel%, C16:0, C18:0, C18:1 cis-9, MUN, mACE, mBHB, and mpbNEFA. Parity 1 cows had less true protein and greater fat percentages than parity 2 and ≥3 cows (% true protein: 3.52, 3.76, 3.81; % fat: 5.55, 4.69, 4.95, for parity 1, 2, ≥3, respectively). De novo and mixed FA rel% were reduced and preformed FA rel% were increased in primiparous compared with parity 2 and ≥3 cows. The increase in preformed FA rel% in primiparous cows agreed with milk markers of energy deficit, such that mpbNEFA, mBHB, and mACE were greatest in parity 1 cows followed by parity ≥3 cows, with parity 2 cows having the lowest concentrations. When measuring milk constituents with FTIR, these results suggest it is critical to account for parity for the majority of estimated milk constituents. We acknowledge the limitation that this study was conducted on a single farm; however, if FTIR technology is to be used as a method of identifying cows maladapted to lactation, understanding variations in early lactation milk constituents is a crucial first step in the practical adoption of this technology.

Diurnal variation of milk fatty acids in early-lactation Holstein cows with and without hyperketonemia.

Estimates of milk constituents by Fourier-transform mid-infrared (FTIR) analysis have been shown to be a useful tool in monitoring energy deficit in early-lactation dairy cows. Our objectives were to describe the diurnal variation in milk fatty acids (FAs) and estimate the association of hyperketonemia with concentrations and diurnal patterns of FTIR estimates of milk FA. Blood samples were collected via jugular catheters bihourly for 5 d from multiparous Holstein cows (n = 28) enrolled between 3 and 9 days in milk. Milk samples were collected thrice daily at 0600, 1400, and 2200 h for d 2, 3, and 4 of the study period. Cows were retrospectively classified as hyperketonemic (HYK; n = 13) or non-HYK (n = 15) based on blood beta-hydroxybutyrate (bBHB) concentrations analyzed during the study period. Cows were classified as HYK if bBHB was ≥ 1.2 mmol/l for ≥ 50% (22/44) of bihourly timepoints; cows were classified as non-HYK if bBHB was ≥ 1.2 mmol/l for < 50% of bihourly timepoints. The HYK cows had bBHB ≥ 1.2 mmol/l for 31.4 ± 6.8 timepoints while the non-HYK cows had bBHB ≥ 1.2 mmol/l for 8.0 ± 3.9 timepoints. We used generalized linear mixed models to analyze concentrations of milk FA over time and differences between HYK groups. The relative percentage of de novo, mixed, and preformed FAs all followed diurnal patterns, however only the yield of preformed FA diurnally cycled, reaching a nadir at 0600 h and peaking at 1400 h. The yield per milking of preformed FA was also greater in the HYK cows than in the non-HYK cows. Oleic acid in milk followed a similar diurnal pattern to the yield of preformed FA, likely driving the cyclical nature of preformed FA. Finally, stearic acid was greater in HYK cows. Our results suggest that FTIR estimates of milk FA offer the potential to provide insight on the energy status of early-lactation cows, and when interested in understanding the absolute concentrations and yields of milk FA, diurnal variation should be considered.

Effects of micellar casein concentrate purity and milk fat on sulfur/eggy flavor in ultrapasteurized milk-based beverages.

Our objectives were to determine the level of milk-derived whey protein (MDWP) removal necessary to achieve no detectable sulfur/eggy flavor in ultrapasteurized fat-free micellar casein concentrate (MCC) beverages (6.5% protein) and in the same beverages containing 1 and 2% milk fat. Micellar casein concentrate with 95% MDWP removal was produced from skim milk (50°C) with a 3×, 3-stage ceramic microfiltration (MF) process using 0.1-µm pore size graded permeability membranes (n = 3). In experiment 1, MCC-based beverages at about 6.5% (wt/wt) true protein were formulated at a fat content of 0.15% fat (wt/wt) at 4 different levels of MDWP removal percentages (95.2%, 91.0%, 83.2%, and 69.3%). In experiment 2, a similar series of beverages at 3 MDWP removal percentages (95.2%, 83.2%, and 69.3%) with 0.1, 1, and 2% fat content were produced. The purity (or completeness of removal of whey protein by MF) of MCC was determined by the Kjeldahl method and sodium dodecyl sulfate (SDS)-PAGE. Sensory properties of beverages were documented by descriptive sensory analysis, and volatile sulfur compounds were evaluated using solid-phase microextraction followed by gas chromatography-triple quadrupole mass spectrometry. The purity of MCC measured by the Kjeldahl method (casein as a percentage of true protein) was higher after thermal treatment than before, whereas MCC purity evaluated by SDS-PAGE was unchanged by heat treatment. The purity of MCC had an effect on the flavor profile of thermally processed beverages at 6.5% protein made with fresh liquid MCC. No sulfur/eggy flavor was detected in MCC beverages when 95% of the MDWP was removed (MCC purity about 93 to 94%) from skim milk by microfiltration at 0.1, 1, and 2% fat. As the fat content of 6.5% protein beverages produced with MCC increased, sulfur/eggy flavor intensity and hydrogen sulfide concentration decreased. However, the effect of increasing milk fat on reducing sulfur/eggy flavor in MCC-based beverages at 6.5% protein was less than that of increasing MDWP removal from MCC. Sulfur off-flavors in neutral-pH dairy protein beverages can be mitigated by use of high-purity MCC or by incorporation of fat in the beverage, or both.

Children's perceptions of fluid milk with varying levels of milkfat.

Schools participating in federal meal programs are limited to serving skim or low-fat (≤1%) flavored and unflavored milk. Few studies have directly addressed child perceptions and preferences for milk containing different amounts of milkfat. The objective of this study was to determine whether children can differentiate between flavored and unflavored fluid milk containing varying levels of milkfat and whether preferences for certain levels of milkfat exist. Flavored and unflavored milks containing 4 different percentages of milkfat (≤0.5, 1, 2, and 3.25%) were high-temperature, short-time processed, filled into half-gallon light-shielded milk jugs, and stored at 4°C in the dark. Milks were evaluated by children (ages 8-13 yr) following 7 d at 4°C. Acceptance testing and tetrad difference testing were conducted on flavored and unflavored milks with and without visual cues to determine if differences were driven by visual or flavor or mouthfeel cues. Child acceptance testing (n = 138 unflavored; n = 123 flavored) was conducted to evaluate liking and perception of selected attributes. Tetrad testing (n = 127 unflavored; n = 129 flavored) was conducted to determine if children could differentiate between different fat levels even in the absence of a difference in acceptance. The experiment was replicated twice. When visual cues were present, children had higher overall liking for 1% and 2% milks than skim for unflavored milk and higher liking for chocolate milks containing at least 1% milk fat than for skim. Differences in liking were driven by appearance, viscosity, and flavor. In the absence of visual cues, no differences were observed in liking or flavor or mouthfeel attributes for unflavored milk but higher liking for at least 1% milk fat in chocolate milk compared with skim was consistent with the presence of visual cues. From tetrad testing, children could visually tell a difference between all unflavored pairs except 2% versus whole milk and could not detect consistent differences between milkfat pairs in the absence of visual cues. For chocolate milk, children could tell a difference between all milk fat pairs with visual cues and could tell a difference between skim versus 2% and skim versus whole milk without visual cues. These results demonstrate that in the absence of package-related flavors, school-age children like unflavored skim milk as well as milk with higher fat content in the absence of visual cues. In contrast, appearance as well as flavor and mouthfeel attributes play a role in children's liking as well as their ability to discriminate between chocolate milks containing different amounts of fat, with chocolate milk containing at least 1% fat preferred. The sensory quality of school lunch milk is vital to child preference, and processing efforts are needed to maximize school milk sensory quality.

Performance evaluation of an enzymatic spectrophotometric method for milk urea nitrogen.

Our objective was to determine the within and between laboratory performance of an enzymatic spectrophotometric method for milk urea nitrogen (MUN) determination. This method first uses urease to hydrolyze urea into ammonia and carbon dioxide. Next, ammonia (as ammonium ions) reacts with 2-oxoglutarate, in the presence of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) and the enzyme glutamate dehydrogenase (GlDH), to form l-glutamic acid, water, and NADP+. The change in light absorption at 340 nm due to the conversion of NADPH to NADP+ is stoichiometrically a function of the MUN content of a milk sample. The relative within (RSDr) and between (RSDR) laboratory method performance values for the MUN enzymatic spectrophotometric method were 0.57% and 0.85%, respectively, when testing individual farm milks. The spectrophotometric MUN method demonstrated better within and between laboratory performance than the International Dairy Federation differential pH MUN method with a much lower RSDr (0.57 vs. 1.40%) and RSDR (0.85 vs. 4.64%). The spectrophotometric MUN method also had similar method performance statistics as other AOAC International official validated chemical methods for primary milk component determinations, with the average of all RSDr and RSDR values being <1%. An official collaborative study of the enzymatic spectrophotometric MUN method is needed to achieve International Dairy Federation, AOAC International, and International Organization for Standardization official method status.

Ready-to-drink protein beverages: Effects of milk protein concentration and type on flavor.

This study evaluated the role of protein concentration and milk protein ingredient [serum protein isolate (SPI), micellar casein concentrate (MCC), or milk protein concentrate (MPC)] on sensory properties of vanilla ready-to-drink (RTD) protein beverages. The RTD beverages were manufactured from 5 different liquid milk protein blends: 100% MCC, 100% MPC, 18:82 SPI:MCC, 50:50 SPI:MCC, and 50:50 SPI:MPC, at 2 different protein concentrations: 6.3% and 10.5% (wt/wt) protein (15 or 25 g of protein per 237 mL) with 0.5% (wt/wt) fat and 0.7% (wt/wt) lactose. Dipotassium phosphate, carrageenan, cellulose gum, sucralose, and vanilla flavor were included. Blended beverages were preheated to 60°C, homogenized (20.7 MPa), and cooled to 8°C. The beverages were then preheated to 90°C and ultrapasteurized (141°C, 3 s) by direct steam injection followed by vacuum cooling to 86°C and homogenized again (17.2 MPa first stage, 3.5 MPa second stage). Beverages were cooled to 8°C, filled into sanitized bottles, and stored at 4°C. Initial testing of RTD beverages included proximate analyses and aerobic plate count and coliform count. Volatile sulfur compounds and sensory properties were evaluated through 8-wk storage at 4°C. Astringency and sensory viscosity were higher and vanillin flavor was lower in beverages containing 10.5% protein compared with 6.3% protein, and sulfur/eggy flavor, astringency, and viscosity were higher, and sweet aromatic/vanillin flavor was lower in beverages with higher serum protein as a percentage of true protein within each protein content. Volatile compound analysis of headspace vanillin and sulfur compounds was consistent with sensory results: beverages with 50% serum protein as a percentage of true protein and 10.5% protein had the highest concentrations of sulfur volatiles and lower vanillin compared with other beverages. Sulfur volatiles and vanillin, as well as sulfur/eggy and sweet aromatic/vanillin flavors, decreased in all beverages with storage time. These results will enable manufacturers to select or optimize protein blends to better formulate RTD beverages to provide consumers with a protein beverage with high protein content and desired flavor and functional properties.

Infrared milk analyzers: Milk urea nitrogen calibration.

Our first objective was to redesign a modified 14-sample milk calibration sample set to obtain a well-distributed range of milk urea nitrogen (MUN) concentrations while maintaining orthogonality with variation in fat, protein, and lactose concentration. Our second objective was to determine the within- and between-laboratory variation in the enzymatic spectrophotometric method on the modified milk calibration samples and degree of uncertainty in MUN reference values, and then use the modified milk calibration samples to evaluate and improve the performance of mid-infrared partial least squares (PLS) models for prediction of MUN concentration in milk. Changes in the modified milk calibration sample formulation and manufacturing procedure were made to achieve the desired range of MUN concentrations. A spectrophotometric enzymatic reference method was used to determine MUN reference values, and the modified milk calibration samples were used to calibrate 3 mid-infrared milk analyzers. The within- and between-laboratory variation in the reference values for MUN were 0.43 and 0.77%, respectively, and the average expanded analytical uncertainty for the mean MUN value of the 14-sample calibration set was (mean ± SD) 16.15 mg/100 g ± 0.09 of milk. After slope and intercept adjustment to achieve a mean difference of zero with the calibration samples, it could be seen that the standard deviation of the differences of predicted versus reference MUN values among 3 different instruments and their PLS models were quite different. The orthogonal sample set was used (1) to determine when a PLS model did not correctly model out the background variation in fat, true protein, or anhydrous lactose; (2) to calculate an intercorrection factor to eliminate that effect, and (3) to improve the model performance (i.e., 50% reduction in standard deviation of the difference between instrument predictions and reference chemistry values for MUN).

The relationship of excessive energy deficit with milk somatic cell score and clinical mastitis.

Periparturient cows go through a period of immune suppression often marked by immune cell dysfunction. Further exacerbation of this dysfunction through early-lactation excessive energy deficit (EED) has been associated with increased susceptibility to infectious conditions such as mastitis. Our objective was to explore the association of milk somatic cell score (SCS) and clinical mastitis (CM) diagnosis in cows identified with EED, diagnosed using each of the following: blood and milk ß-hydroxybutyrate (BHB), milk predicted blood nonesterified fatty acid (mpbNEFA) concentrations, or milk de novo fatty acid (FA) relative percentages (rel %). We analyzed data collected from 396 multiparous Holstein cows from 2 New York farms in a prospective cohort study. Coccygeal vessel blood samples and composite milk samples were collected twice weekly from 3 to 18 days in milk (DIM) for a total of 4 time points per cow (T1, T2, T3, T4). Blood was analyzed using a hand-held meter, and milk was analyzed using Fourier-transform mid-infrared spectrometry for milk BHB and mpbNEFA concentrations, milk de novo FA rel %, and somatic cell count. Excessive energy deficit was diagnosed as blood BHB ≥ 1.2 mmol/L, milk BHB ≥ 0.14 mmol/L, mpbNEFA ≥ 0.55 mmol/L, or de novo FA ≤ 22.7 rel %, depending on the model. Clinical mastitis cultures were collected from 4 to 60 DIM by on-farm personnel. Incidence of hyperketonemia as determined by blood BHB was 13.4%, and incidence of CM was 23.9%. Separate repeated-measures ANOVA models were developed for each EED diagnostic analyte for parity groups 2, 3, and ≥4 to assess differences in SCS; t-test analyses were similarly used to assess the association of each diagnostic analyte with CM at each time point. For all diagnostic analytes, apart from milk BHB, cows diagnosed with EED tended to have lower SCS than their non-EED counterparts. This was especially apparent at T1 for all parity groups, and at T2, T3, and T4 for blood BHB and mpbNEFA. For EED diagnosis via mpbNEFA, mean SCS were lower in parity ≥4, with a difference in mean SCS between EED and non-EED animals of 0.7 SCS units, equating to a somatic cell count in EED animals approaching half that of non-EED (EED = 67,000 cells/mL, non-EED = 107,000 cell/mL). No important relationships were observed between CM diagnosis and blood BHB, milk BHB, or mpbNEFA. For de novo FA rel %, reductions in this analyte were noted before CM diagnosis at all time points. Although the relationship between EED and CM is still unclear, our findings suggest that cows in EED, diagnosed using blood BHB or mpbNEFA during the first 18 DIM, have a tendency toward lower SCS compared with their non-EED counterparts.

Effect of hyperketonemia on the diurnal patterns of energy-related blood metabolites in early-lactation dairy cows.

Most dairy cows experience a period of energy deficit in early lactation, resulting in increased plasma concentrations of nonesterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB). Our objectives were to determine (1) the diurnal variation in plasma BHB and NEFA, (2) the correlation between plasma NEFA and BHB when accounting for diurnal changes, and (3) the effect of hyperketonemia (HYK) on the diurnal pattern of blood metabolites. Jugular catheters were placed in 28 multiparous Holstein cows between 3 and 9 days in milk, and blood samples were collected every 2 h for 96 h. Cows were retrospectively classified as HYK positive (HYK; n = 13) if they had plasma BHB concentrations ≥1.2 mmol/L for ≥3 study days, or HYK negative (non-HYK; n = 15) if they had plasma BHB concentrations ≥1.2 mmol/L for ≤2 study days. Generalized linear mixed models were used to analyze concentrations of analytes over time and differences in metabolites between HYK groups. The correlation between total plasma NEFA and BHB was analyzed by calculating the area under the curve for plasma NEFA and BHB for all cows. Plasma NEFA reached a peak approximately 2 h before morning feed delivery, falling to a nadir in the late evening. Plasma BHB was at a nadir at the time of morning feed delivery, peaking 4 h later. We observed a strong positive correlation between daily plasma NEFA and BHB. Additionally, HYK cows had greater concentrations of plasma NEFA and BHB than non-HYK cows. The HYK cows also experienced a greater magnitude of change in BHB throughout the day than the non-HYK cows. Our results suggest that the time relative to feeding should be considered when analyzing plasma metabolites, as classification of energy status may change throughout a day.

Association of mid-infrared-predicted milk and blood constituents with early-lactation disease, removal, and production outcomes in Holstein cows.

Partial least squares regression estimates of milk and blood constituents using Fourier-transform mid-infrared (FTIR) analysis have shown promise as a tool for monitoring early-lactation excessive energy deficit in dairy herds. Our objective was to analyze milk via FTIR to determine the association of early-lactation predicted milk ß-hydroxybutyrate (BHB) concentrations, predicted blood nonesterified fatty acid (NEFA) concentrations, and predicted milk de novo fatty acid (FA) percentages relative to total FA concentrations, with the risk of disease or removal in early lactation (hyperketonemia, displaced abomasum, metritis, culling, or death) and average daily milk yield during the first 15 wk of lactation. We enrolled 517 multiparous Holstein cows from 2 dairy farms in New York. Composite milk samples were collected twice weekly from 3 to 18 DIM for a total of 4 timepoints (T1, T2, T3, T4) and analyzed using FTIR spectrometry for milk BHB and FA composition and predicted blood NEFA. Blood samples were collected for hyperketonemia determination (BHB ≥ 1.2 mmol/L) using a handheld meter, and farm-diagnosed occurrence of disease or removal during the first 30 DIM and average daily milk yield during the first 15 wk of lactation were collected from herd management software. The incidence of disease or removal between 3 and 18 DIM was 20.2%. Explanatory models for disease or removal were developed for each predicted constituent of interest at each timepoint using fixed-effect multivariable Poisson regression. Repeated measures ANOVA models were developed for each predicted constituent to assess differences in average daily milk yield. For all timepoints, increased risk of disease or removal was associated with higher predicted milk BHB [relative risk (RR)T1 = 2.0; RRT2 = 3.4; RRT3 = 5.2; RRT4 = 9.1], higher predicted blood NEFA (RRT1 = 2.7; RRT2 = 2.5; RRT3 = 3.8; RRT4 = 10.0), and lower predicted milk de novo FA relative percentages (RRT1 = 2.9; RRT2 = 3.3; RRT3 = 5.8; RRT4 = 7.2). Average daily milk yield was increased for cows above the cut point for predicted milk BHB (2.1 kg/d) and predicted blood NEFA (3.5 kg/d) and below the cut point for de novo FA relative percentages (2.3 kg/d). Our results suggest that FTIR-predicted milk BHB, blood NEFA, and milk de novo FA relative percentages are promising indicators of subsequent disease or removal in early lactation; their positive relationship with milk yield warrants further exploration.

Identification of the source of volatile sulfur compounds produced in milk during thermal processing.

Volatile sulfur compounds in ultra-pasteurized (UP) milk are the major contributors to sulfur/burnt and eggy flavors, and these flavors are disliked by consumers. Previous research has established distinct differences in flavor profiles of fluid milk processed by high temperature, short time pasteurization (HTST) and UP by direct steam injection (DSI-UP) or indirect heat (IND-UP). An understanding of the contribution of the individual milk proteins to sulfur off-flavors would clarify the source of sulfur flavors in UP milks. The objective of this study was to determine the source of volatile sulfur compounds in fluid milk with a specific focus on the comparison of heat treatment effects on milks by HTST and UP. Formulated skim milks (FSM) were manufactured by blending micellar casein concentrate and serum protein isolate (SPI). Three different caseins as a percentage of true protein (FSM95, FSM80, and FSM60) were formulated to determine the source of sulfur/burnt and eggy flavors. Freshly processed micellar casein concentrate or SPI were blended to achieve a true protein content of about 3.2%. Raw skim milk served as a control. Skim milk and FSM were pasteurized at 78°C for 15 s (HTST) or 140°C for 2.3 s by IND-UP or DSI-UP. The experiment was replicated twice. Sensory properties of milks and FSM were documented by descriptive sensory analysis. Volatile sulfur compounds in milks and FSM were evaluated using solid-phase microextraction followed by gas chromatography-triple quadrupole mass spectrometry combined with a sulfur selective flame photometric detector. The FSM with higher SPI as a percent of true protein had higher sensory sulfur/burnt and eggy flavors along with elevated concentrations of hydrogen sulfide and carbon disulfide compared with skim milk or FSM with lower proportions of SPI. Sulfur compounds including dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, dimethyl sulfoxide, and methional were not associated with sulfur/burnt and eggy flavors, which suggests that these compounds may not specifically contribute to the sulfur/burnt and eggy off-flavors of UP milks. Sensory panelists found higher overall aroma impact, and cooked, sulfur/burnt, and eggy flavors for DSI-UP, followed by IND-UP and HTST. The combination of sensory and instrumental methods used in the current study effectively identified that milk serum proteins are the source of sulfur compounds in milk, and further confirmed the contribution of hydrogen sulfide and carbon disulfide to eggy and sulfur/burnt flavors, respectively.

Effects of feeding synthetic zeolite A during the prepartum period on serum mineral concentration, oxidant status, and performance of multiparous Holstein cows.

The objective of this study was to determine the effects of feeding synthetic zeolite A for 3 wk before expected calving on peripartal serum mineral concentrations, hypocalcemia, oxidant status, and performance. Holstein cows (n = 55) entering their second or greater lactations were assigned randomly to 1 of 2 dietary treatments starting 21 d before expected calving: control (CON: 40% corn silage, 33% wheat straw, and 27% concentrate; n = 29) or experimental [EXP: CON plus zeolite A (X-Zelit, Protekta Inc., Lucknow, ON, Canada/Vilofoss, Graasten, Denmark; n = 26) at an inclusion rate of 3.3% of dry matter, targeting 500 g/d as-fed]. Cows were fed the same postpartum diet and housed in individual tiestalls through 28 d in milk. Cows fed EXP had higher serum Ca concentrations as parturition approached and during the immediate postpartum period. Serum P concentrations were lower for the EXP-fed cows during the prepartum period and the first 2 d of lactation, whereas serum Mg concentrations were lower than those of the CON-fed cows only during the immediate periparturient period. Cows fed EXP had decreased prevalence of subclinical hypocalcemia (SCH) from d -1 through 3 relative to day of parturition, with the largest difference occurring within the first day postpartum. Prepartum dry matter intake tended to be decreased and rumination was decreased in cows fed EXP; however; postpartum dry matter intake, rumination, milk yield, milk component yield, and colostrum measurements did not differ between treatments. Cows fed EXP tended to have increased hazard of pregnancy by 150 d in milk when controlling for parity compared with CON-fed cows; potential reproductive benefits merit further study. This study demonstrated that zeolite A supplementation during the prepartum period results in markedly improved serum Ca concentrations around parturition and similar postpartum performance compared with controls and is effective at decreasing hypocalcemia in multiparous Holstein cows.

Does vitamin fortification affect light oxidation in fluid skim milk?

Off-flavors in milk related to light oxidation form due to photoxidation of native riboflavin and tetrapyrroles, resulting in an array of lipid oxidation compounds. Recent work has established that fortification with water-dispersible vitamin A can result in off-flavors in fluid skim milk caused by vitamin A degradation products in the vitamin premix. The objective of this study was to determine the role of vitamin fortification on light oxidation of high temperature, short time pasteurized fluid skim milk. First, the aroma profiles and aroma-active volatile compounds in light-exposed vitamin premixes were determined by exposing the premixes to fluorescent (FL) or light-emitting diode (LED) light at 2,000 lx at 4°C for 0, 2, 4, 8, or 24 h. A trained panel (n = 6) documented aroma profiles of each vitamin premix at each time point. Headspace solid-phase microextraction followed by gas chromatography-olfactometry and gas chromatography-mass spectrometry (GC-MS) were performed to characterize aroma-active compounds in light-exposed vitamin premixes. In the second experiment, commercial vitamin premixes (vitamin A and vitamin D in oil and water matrices) were used to fortify skim milk (vitamin A: 3,000 IU/946 mL; vitamin D: 600 IU/946 mL). Skim milk was pasteurized, homogenized, and packaged in 946-mL high-density polyethylene jugs. Milks were exposed to FL or LED light at 2,000 lx at 4°C for 4, 12, 24, or 48 h. Controls with and without vitamins and light shielding were included. Riboflavin and vitamin A and D degradation were quantified via ultra-high-performance liquid chromatography. A trained panel (n = 8) documented sensory profiles of milks at each time point. Lipid oxidation volatile compounds were quantified via solid-phase microextraction with GC-MS. Vitamin degradation volatile compounds were quantified via solvent-assisted sorptive stir bar extraction with GC-MS. Riboflavin, vitamin A, and vitamin D degradation were consistent with that reported in previous studies. We found no effect of vitamin fortification on development of typical light oxidation-related off-flavors (cardboard and mushroom) or lipid oxidation-related volatiles (hexanal and heptanal). A perfumey/floral flavor was documented in the oil-based vitamin A-fortified milk, suggesting that light exposure affected the off-flavors contributed by water- versus oil-based vitamin fortification. These results show no evidence that vitamin fortification at current levels provides any protection against light oxidation-related off-flavors in fluid milk.

Flavor and flavor chemistry differences among milks processed by high-temperature, short-time pasteurization or ultra-pasteurization.

Typical high-temperature, short-time (HTST) pasteurization encompasses a lower heat treatment and shorter refrigerated shelf life compared with ultra-pasteurization (UP) achieved by direct steam injection (DSI-UP) or indirect heat (IND-UP). A greater understanding of the effect of different heat treatments on flavor and flavor chemistry of milk is required to characterize, understand, and identify the sources of flavors. The objective of this study was to determine the differences in the flavor and volatile compound profiles of milk subjected to HTST, DSI-UP, or IND-UP using sensory and instrumental techniques. Raw skim and raw standardized 2% fat milks (50 L each) were processed in triplicate and pasteurized at 78°C for 15 s (HTST) or 140°C for 2.3 s by DSI-UP or IND-UP. Milks were cooled and stored at 4°C, then analyzed at d 0, 3, 7, and 14. Sensory attributes were determined using a trained panel, and aroma active compounds were evaluated by solid-phase micro-extraction or stir bar sorptive extraction followed by gas chromatography-mass spectrometry, gas chromatography-olfactometry, and gas chromatography-triple quad mass spectrometry. The UP milks had distinct cooked and sulfur flavors compared with HTST milks. The HTST milks had less diversity in aroma active compounds compared with UP milks. Flavor intensity of all milks decreased by d 14 of storage. Aroma active compound profiles were affected by heat treatment and storage time in both skim and 2% milk. High-impact aroma active compounds were hydrogen sulfide, dimethyl trisulfide, and methional in DSI-UP and 2 and 3-methylbutanal, furfural, 2-heptanone, 2-acetyl-1-pyrroline, 2-aminoacetophenone, benzaldehyde, and dimethyl sulfide in IND-UP. These results provide a foundation knowledge of the effect of heat treatments on flavor development and differences in sensory quality of UP milks.

Management practices, physically effective fiber, and ether extract are related to bulk tank milk de novo fatty acid concentration on Holstein dairy farms.

The objective of this study was to evaluate the relationship of management practices and dietary factors with de novo fatty acid concentration in bulk tank milk from commercial dairy farms milking Holstein cows. Farms were selected based on de novo fatty acid concentration during the 6 mo before the farm visit and were categorized as high de novo (HDN; 24.61 ± 0.75 g/100 g of fatty acids, mean ± standard deviation; n = 19) or low de novo (LDN; 23.10 ± 0.88 g/100 g of fatty acids; n = 20). Farms were visited once in February, March, or April 2015 and evaluated based on management and facility design known to affect cow behavior, physical and chemical characteristics of the diet, and ration formulation and forage analyses obtained from the farm's nutritionist. We observed no differences between HDN and LDN farms in farm size, time away from the pen for milking, days in milk, or body condition score. We detected no differences between HDN and LDN farms in milk fat or true protein yield; however, milk fat and protein content and de novo fatty acid yield per day were higher for HDN farms, as was gross income per unit of milk sold. High de novo farms tended to be more likely to deliver fresh feed twice versus once per day, have a freestall stocking density ≤110%, and provide ≥46 cm of feed bunk space per cow. We observed no detectable differences in forage quality or ration dry matter, crude protein, or starch content. However, ether extract was lower and physically effective neutral detergent fiber was higher for HDN farms. Feeding management, stocking density, dietary ether extract content, and the physical characteristics of the diet are related to de novo fatty acid, fat, and protein concentration in bulk tank milk from high-producing Holstein dairy farms.

The effect of vitamin concentrates on the flavor of pasteurized fluid milk.

Fluid milk consumption in the United States continues to decline. As a result, the level of dietary vitamin D provided by fluid milk in the United States diet has also declined. Undesirable flavor(s)/off flavor(s) in fluid milk can negatively affect milk consumption and consumer product acceptability. The objectives of this study were to identify aroma-active compounds in vitamin concentrates used to fortify fluid milk, and to determine the influence of vitamin A and D fortification on the flavor of milk. The aroma profiles of 14 commercial vitamin concentrates (vitamins A and D), in both oil-soluble and water-dispersible forms, were evaluated by sensory and instrumental volatile compound analyses. Orthonasal thresholds were determined for 8 key aroma-active compounds in skim and whole milk. Six representative vitamin concentrates were selected to fortify skim and 2% fat pasteurized milks (vitamin A at 1,500-3,000 IU/qt, vitamin D at 200-1,200 IU/qt, vitamin A and D at 1,000/200-6,000/1,200 IU/qt). Pasteurized milks were evaluated by sensory and instrumental volatile compound analyses and by consumers. Fat content, vitamin content, and fat globule particle size were also determined. The entire experiment was done in duplicate. Water-dispersible vitamin concentrates had overall higher aroma intensities and more detected aroma-active compounds than oil-soluble vitamin concentrates. Trained panelists and consumers were able to detect flavor differences between skim milks fortified with water-dispersible vitamin A or vitamin A and D, and unfortified skim milks. Consumers were unable to detect flavor differences in oil-soluble fortified milks, but trained panelists documented a faint carrot flavor in oil-soluble fortified skim milks at higher vitamin A concentrations (3,000-6,000 IU). No differences were detected in skim milks fortified with vitamin D, and no differences were detected in any 2% milk. These results demonstrate that vitamin concentrates may contribute to off flavor(s) in fluid milk, especially in skim milk fortified with water-dispersible vitamin concentrates.

The influence of ultra-pasteurization by indirect heating versus direct steam injection on skim and 2% fat milks.

Fluid milk is traditionally pasteurized by high temperature, short time (HTST) pasteurization, which requires heating to at least 72°C for 15 s. Ultra-pasteurization (UP) extends milk shelf life and is defined as heating to at least 138°C for 2 s. The UP process can be done by indirect heating (IND) or by direct steam injection (DSI). The influence of these 2 UP methods on milk flavor has not been widely investigated. The objective of this study was to compare the effect of HTST, IND-UP, and DSI-UP on sensory perception of fluid milk. Raw skim and standardized 2% milks were pasteurized at 140°C for 2.3 s by IND or DSI or by HTST (78°C, 15 s) and homogenized at 20.7 MPa. The processed milks were stored in light-shielded opaque high-density polyethylene containers at 4°C and examined by descriptive analysis and microbial analysis on d 3, 7, and 14. Furosine and serum protein denaturation analyses were performed on d 0 and 14 as an indicator of heat treatment. Last, consumer acceptance testing was conducted at d 10, with adults (n = 250) and children (ages 8 to13 y, n = 100) who were self-reported consumers of skim or 2% milk; consumers only received samples for either skim or 2% milk. The entire experiment was repeated in triplicate. Milks treated by HTST had lower cooked flavor than either UP milk. Milks heated by DSI-UP were characterized by sulfur or eggy and cooked flavors, whereas IND-UP milks had higher sweet aromatic and sweet taste compared with DSI-UP milk. Aromatic flavor intensities of all milks decreased across 14 d of storage. Furosine concentrations and serum protein denaturation were highest for the IND treatments, followed by DSI and HTST. Furosine content in both skim and 2% milk increased with time, but the increase was faster in IND-UP skim milk. Adult and child consumers preferred HTST milk over either UP milk, regardless of fat content. Ultra-pasteurization by IND or DSI did not affect consumer acceptance at 10 d postprocessing, but traditional HTST milks were preferred by consumers of all ages.