From cow to cheese: Novel phenotypes related to the sensory profile of model cheeses from individual cows.

Milk samples were taken once from a total of 1,224 Brown Swiss cows from 83 herds, and 1,500 mL of raw full-fat milk from each cow was processed according to a laboratory-scale model-cheese-making procedure. A sensory panel was assembled and the members trained to evaluate the sensory profile of individual model cheeses. The protocol scorecard was composed of 7 main sensory descriptors related to smell intensity, flavor intensity, taste (salt and sour), and texture (elasticity, firmness, and moisture), and 40 sensory attributes describing smell and flavor profiles. Sensory data were analyzed using a mixed model that included random effects of herd, animal, and panelist, as well as fixed effects of dairy system, days in milk, parity, and order of cheese presentation, and covariates for cheese weight and fat:protein ratio. The sensory profile was not much affected by the dairy farming systems included in the trial, but it was affected by farm within dairy system: cheeses from traditional dairy farms had a greater wood/humus attribute of both smell and flavor than those from modern farm. Of the modern farms, cheeses from those using total mixed rations including silages had a more intense smell of sour milk and a firmer, less moist texture than those using total mixed rations without silages. Moreover, for all the sensory traits, we found less variance related to herd and animals than that related to the panelists and the residuals. Stage of lactation was found to be the most important, whereas parity was not relevant. In particular, cheese smell intensity (and some related attributes) exhibited a quadratic trend with lower values in mid-lactation, whereas flavor and salt descriptors were more intense in the last period of lactation.

Breed of cow and herd productivity affect milk composition and modeling of coagulation, curd firming, and syneresis.

Milk coagulation properties (MCP) have been widely investigated in the past using milk collected from different cattle breeds and herds. However, to our knowledge, no previous studies have assessed MCP in individual milk samples from several multi-breed herds characterized by either high or low milk productivity, thereby allowing the effects of herd and cow breed to be evaluated independently. Multi-breed herds (n=41) were classified into 2 categories based on milk productivity (high vs. low), defined according to the average milk net energy yielded daily by lactating cows. Milk samples were taken from 1,508 cows of 6 different breeds: 3 specialized dairy (Holstein-Friesian, Brown Swiss, Jersey) and 3 dual-purpose (Simmental, Rendena, Alpine Grey) breeds, and analyzed in duplicate (3,016 tests) using 2 lactodynamographs to obtain 240 curd firming (CF) measurements over 60min (1 every 15 s) for each duplicate. The 5 traditional single-point MCP (RCT, k20, a30, a45, and a60) were yielded directly by the instrument from the available CF measures. All 240 CF measures of each replicate were also used to estimate 4 individual equation parameters: RCT estimated according to curd firm change over time modeling (RCTeq), asymptotic potential curd firmness (CFP), curd firming instant rate constant (kCF), and syneresis instant rate constant (kSR) and 2 derived traits: maximum curd firmness achieved within 45min (CFmax) and time at achievement of CFmax (tmax) by curvilinear regression using a nonlinear procedure. Results showed that the effect of herd-date on traditional and modeled MCP was modest, ranging from 6.1% of total variance for k20 to 10.7% for RCT, whereas individual animal variance was the highest, ranging from 32.0% for tmax to 82.5% for RCTeq. The repeatability of MCP was high (>80%) for all traits except those associated with the last part of the lactodynamographic curve (i.e., a60, kSR, kCF, and tmax: 57 to 71%). Reproducibility, taking into account the effect of instrument, was equal to or slightly lower than repeatability. Milk samples collected in farms characterized by high productivity exhibited delayed coagulation (RCTeq: 18.6 vs. 16.3min) but greater potential curd firmness (CFP: 76.8 vs. 71.9mm) compared with milk samples collected from low-productivity herds. Parity and days in milk influenced almost all MCP. Large differences in all MCP traits were observed among breeds, both between specialized and dual-purpose breeds and within these 2 groups of breeds, even after adjusting for milk quality and yield. Milk quality and MCP of samples from Jersey cows, and coagulation time of samples from Rendena cows were better than in milk from Holstein-Friesian cows, and intermediate results were found with the other breeds of Alpine origin. The results of this study, taking into account the intrinsic limitation of this technique, show that the effects of breed on traditional and modeled MCP are much greater than the effects of herd productivity class, parity, and DIM. Moreover, the variance in individual animals is much greater than the variance in individual herds within herd productivity class. It seems that improvement in MCP depends more on genetics (e.g., breed, selection) than on environmental and management factors.

Genetic analysis of coagulation properties, curd firming modeling, milk yield, composition, and acidity in Sarda dairy sheep.

Sheep milk is an important source of food, especially in Mediterranean countries, and is used in large part for cheese production. Milk technological traits are important for the sheep dairy industry, but research is lacking into the genetic variation of such traits. Therefore the aim of this study was to estimate the heritability of traditional milk coagulation properties and curd firmness modeled on time t (CFt) parameters, and their genetic relationships with test-day milk yield, composition (fat, protein, and casein content), and acidity in Sarda dairy sheep. Milk samples from 1,121 Sarda ewes from 23 flocks were analyzed for 5 traditional coagulation properties by lactodynamographic tests conducted for up to 60min: rennet coagulation time (min), curd-firming time (k20, min), and 3measures of curd firmness (a30, a45, and a60, mm). The 240 curd firmness observations (1 every 15 s) from each milk sample were recorded, and 4 parameters for each individual sample equation were estimated: rennet coagulation time estimated from the equation (RCTeq), the asymptotic potential curd firmness (CFP), the curd firming instant rate constant (kCF), and the syneresis instant rate constant (kSR). Two other derived traits were also calculated (CFmax, the maximum curd firmness value; and tmax, the attainment time). Multivariate analyses using Bayesian methodology were performed to estimate the genetic relationships of milk coagulation properties and CFt with the other traits; statistical inference was based on the marginal posterior distributions of the parameters of concern. The marginal posterior distribution of heritability estimates of milk yield (0.16±0.07) and composition (0.21±0.11 to 0.28±0.10) of Sarda ewes was similar to those often obtained for bovine species. The heritability of rennet coagulation time as a single point trait was also similar to that frequently obtained for cow milk (0.19±0.09), whereas the same trait calculated as an individual equation parameter exhibited larger genetic variation and a higher heritability estimate (0.32±0.11). The other curd firming and syneresis traits, whether as traditional single point observations or as individual equation parameters and derived traits, were characterized by heritability estimates lower than for coagulation time and for the corresponding bovine milk traits (0.06 to 0.14). Phenotypic and additive genetic correlations among the 11 technological traits contribute to describing the interdependencies and meanings of different traits. The additive genetic relationships of these technological traits with the single test-day milk yield and composition were variable and showed milk yield to have unfavorable effects on all measures of curd firmness (a30, a45, a60, CFP, and CFmax) and tmax, but favorable effects on both instant rate constants (kCF and kSR). Milk fat content had a positive effect on curd firmness traits, especially on those obtained from CFt equations, whereas the negative effects on both coagulation time traits were attributed to the milk protein and casein contents. Finally, in view of the estimated heritabilities and additive genetic correlations, enhancement of technological traits of sheep milk through selective breeding could be feasible in this population.

Prediction and repeatability of milk coagulation properties and curd-firming modeling parameters of ovine milk using Fourier-transform infrared spectroscopy and Bayesian models.

The aim of this study was to apply Bayesian models to the Fourier-transform infrared spectroscopy spectra of individual sheep milk samples to derive calibration equations to predict traditional and modeled milk coagulation properties (MCP), and to assess the repeatability of MCP measures and their predictions. Data consisted of 1,002 individual milk samples collected from Sarda ewes reared in 22 farms in the region of Sardinia (Italy) for which MCP and modeled curd-firming parameters were available. Two milk samples were taken from 87 ewes and analyzed with the aim of estimating repeatability, whereas a single sample was taken from the other 915 ewes. Therefore, a total of 1,089 analyses were performed. For each sample, 2 spectra in the infrared region 5,011 to 925 cm-1 were available and averaged before data analysis. BayesB models were used to calibrate equations for each of the traits. Prediction accuracy was estimated for each trait and model using 20 replicates of a training-testing validation procedure. The repeatability of MCP measures and their predictions were also compared. The correlations between measured and predicted traits, in the external validation, were always higher than 0.5 (0.88 for rennet coagulation time). We confirmed that the most important element for finding the prediction accuracy is the repeatability of the gold standard analyses used for building calibration equations. Repeatability measures of the predicted traits were generally high (≥95%), even for those traits with moderate analytical repeatability. Our results show that Bayesian models applied to Fourier-transform infrared spectra are powerful tools for cheap and rapid prediction of important traits in ovine milk and, compared with other methods, could help in the interpretation of results.

Genome-wide association study for cheese yield and curd nutrient recovery in dairy cows.

Cheese production and consumption are increasing in many countries worldwide. As a result, interest has increased in strategies for genetic selection of individuals for technological traits of milk related to cheese yield (CY) in dairy cattle breeding. However, little is known about the genetic background of a cow's ability to produce cheese. Recently, a relatively large panel (1,264 cows) of different measures of individual cow CY and milk nutrient and energy recoveries in the cheese (REC) became available. Genetic analyses showed considerable variation for CY and for aptitude to retain high proportions of fat, protein, and water in the coagulum. For the dairy industry, these characteristics are of major economic importance. Nevertheless, use of this knowledge in dairy breeding is hampered by high costs, intense labor requirement, and lack of appropriate technology. However, in the era of genomics, new possibilities are available for animal breeding and genetic improvement. For example, identification of genomic regions involved in cow CY might provide potential for marker-assisted selection. The objective of this study was to perform genome-wide association studies on different CY and REC measures. Milk and DNA samples from 1,152 Italian Brown Swiss cows were used. Three CY traits expressing the weight (wt) of fresh curd (%CYCURD), curd solids (%CYSOLIDS), and curd moisture (%CYWATER) as a percentage of weight of milk processed, and 4 REC (RECFAT, RECPROTEIN, RECSOLIDS, and RECENERGY, calculated as the % ratio between the nutrient in curd and the corresponding nutrient in processed milk) were analyzed. Animals were genotyped with the Illumina BovineSNP50 Bead Chip v.2. Single marker regressions were fitted using the GenABEL R package (genome-wide association using mixed model and regression-genomic control). In total, 103 significant associations (88 single nucleotide polymorphisms) were identified in 10 chromosomes (2, 6, 9, 11, 12, 14, 18, 19, 27, 28). For RECFAT and RECPROTEIN, high significance peaks were identified in Bos taurus autosome (BTA) 6 and BTA11, respectively. Marker ARS-BFGL-NGS-104610 (∼104.3 Mbp) was highly associated with RECPROTEIN and Hapmap52348-rs29024684 (∼87.4 Mbp), closely located to the casein genes on BTA6, with RECFAT. Genomic regions identified may enhance marker-assisted selection in bovine cheese breeding beyond the use of protein (casein) and fat contents, whereas new knowledge will help to unravel the genomic background of a cow's ability for cheese production.

The nonlinear effect of somatic cell count on milk composition, coagulation properties, curd firmness modeling, cheese yield, and curd nutrient recovery.

The aim of this study was to investigate the relationships between somatic cell count (SCC) in milk and several milk technological traits at the individual cow level. In particular, we determined the effects of very low to very high SCC on traits related to (1) milk yield and composition; (2) coagulation properties, including the traditional milk coagulation properties (MCP) and the new curd firming model parameters; and (3) cheese yield and recovery of milk nutrients in the curd (or loss in the whey). Milk samples from 1,271 Brown Swiss cows from 85 herds were used. Nine coagulation traits were measured: 3 traditional MCP [rennet coagulation time (RCT, min), curd firming rate (k20, min), and curd firmness after 30 min (a30, mm)] and 6 new curd firming and syneresis traits [potential asymptotic curd firmness at infinite time (CFP, mm), curd firming instant rate constant (kCF, % × min(-1)), syneresis instant rate constant (kSR, % × min(-1)), rennet coagulation time estimated using the equation (RCTeq, min), maximum curd firmness achieved within 45 min (CFmax, mm), and time at achievement of CFmax (tmax, min)]. The observed cheese-making traits included 3 cheese yield traits (%CYCURD, %CYSOLIDS, and %CYWATER, which represented the weights of curd, total solids, and water, respectively, as a percentage of the weight of the processed milk) and 4 nutrient recoveries in the curd (RECFAT, RECPROTEIN, RECSOLIDS, and RECENERGY, which each represented the percentage ratio between the nutrient in the curd and milk). Data were analyzed using a linear mixed model with the fixed effects of days in milk, parity, and somatic cell score (SCS), and the random effect of herd-date. Somatic cell score had strong influences on casein number and lactose, and also affected pH; these were traits characterized by a quadratic pattern of the data. The results also showed a negative linear relationship between SCS and milk yield. Somatic cell score influenced almost all of the tested coagulation traits (both traditional and modeled), with the exceptions of k20, CFP, and kSR. Gelation was delayed when the SCS decreased (slightly) and when it increased (strongly) with respect to a value of 2, as confirmed by the quadratic patterns observed for both RCT and RCTeq. The SCS effect on a30 showed a quadratic pattern almost opposite to that observed for RCT. With respect to the CFt parameters, kCF decreased linearly as SCS increased, resulting in a linear decrease of CFmax and a quadratic pattern for tmax. Milk SCS attained significance for %CYCURD, %CYWATER, and RECPROTEIN. As the SCS increased beyond 3, we observed a progressive quadratic decrease of the water retained in the curd (%CYWATER), which caused a parallel decrease in %CYCURD. With respect to RECPROTEIN, the negative effect of SCS was almost linear. Recovery of fat and (consequently) RECENERGY was characterized by a more evident quadratic trend, with the most favorable values associated with an intermediate SCS. Together, our results confirmed that high SCS has a negative effect on milk composition and technological traits, highlighting the nonlinear trends of some traits across the different classes of SCS. Moreover, we report that a very low SCS has a negative effect on some technological traits of milk.

The 9-MilCA method as a rapid, partly automated protocol for simultaneously recording milk coagulation, curd firming, syneresis, cheese yield, and curd nutrients recovery or whey loss.

The aim of this study was to propose and test a new laboratory cheesemaking procedure [9-mL milk cheesemaking assessment (9-MilCA)], which records 15 traits related to milk coagulation, curd firming, syneresis, cheese yield, and curd nutrients recovery or whey loss. This procedure involves instruments found in many laboratories (i.e., heaters and lacto-dynamographs), with an easy modification of the sample rack for the insertion of 10-mL glass tubes. Four trials were carried out to test the 9-MilCA procedure. The first trial compared 8 coagulation and curd firming traits obtained using regular or modified sample racks to process milk samples from 60 cows belonging to 5 breeds and 3 farms (480 tests). The obtained patterns exhibited significant but irrelevant between-procedure differences, with better repeatability seen for 9-MilCA. The second trial tested the reproducibility and repeatability of the 7 cheesemaking traits obtained using the 9-MilCA procedure on individual samples from 60 cows tested in duplicate in 2 instruments (232 tests). The method yielded very repeatable outcomes for all 7 tested cheese yield and nutrient recovery traits (repeatability >98%), with the exception of the fresh cheese yield (84%), which was affected by the lower repeatability (67%) of the water retained in the curd. In the third trial (96 tests), we found that using centrifugation in place of curd cooking and draining (as adopted in several published studies) reduced the efficiency of whey separation, overestimated all traits, and worsened the repeatability. The fourth trial compared 9-MilCA with a more complex model cheese-manufacturing process that mimics industry practices, using 1,500-mL milk samples (72 cows, 216 tests). The average results obtained from 9-MilCA were similar to those obtained from the model cheeses, with between-method correlations ranging from 78 to 99%, except for the water retained in the curd (r=54%). Our results indicate that new 9-MilCA method is a powerful research tool that allows the rapid, inexpensive, and partly automated analysis processing 40 samples per day with 2 replicates each, using 1 lacto-dynamograph, 2 heaters, and 3 modified sample racks, and yields a complete picture of the cheesemaking process (e.g., milk gelation, curd firming, syneresis, and whey expulsion) as well as the cheese yield and the efficiency of energy or nutrients retention in the cheese or loss in the whey.

The influence of dietary nitrogen reduction and conjugated linoleic acid supply to dairy cows on fatty acids in milk and their transfer to ripened cheese.

The aim of this study was to investigate the consequences of reducing the dietary crude protein content, with or without a supply of protected conjugated linoleic acid (CLA), on the milk fatty acid (FA) yield and recovery in 90d ripened cheese. Twenty mid-lactation Friesian dairy cows were reared for 4 periods of 3wk each in groups of 5, following a 4×4 Latin square design. Cows were fed 4 different rations, consisting of a combination of the 2 dietary crude protein levels [150 (CP15) or 123 (CP12) g of crude protein/kg of dry matter], with or without a conjugated linoleic acid supply (80g/d, providing 5.57 and 5.40g/d of C18:2 cis-9,trans-11 and C18:2 trans-10,cis-12, respectively). Milk yield was recorded. Twice in each period, milk samples were analyzed for protein, fat, and lactose content, and 10 L milk samples (pooled by group) were processed to produce 96 cheeses, which were ripened for 90d. Milk and cheese fat were analyzed for their FA profiles. Milk and cheese FA were expressed as daily yields and relative proportions, and nutrient recoveries were computed. Dietary crude protein reduction had small or no effects on the yield and relative presence of FA in milk and cheese, except for a small increase in mid-chain branched saturated fatty acids. The CLA supply strongly reduced the yield of various categories of FA, and had major effects on short-chain FA of de novo synthesis, leading to changes in the relative proportions of the various FA in milk and cheese. The addition of CLA tended to reduce uniformly the recovery of all milk constituents and of short-, medium-, and long-chain FA groups, but we observed large differences among individual FA with apparent recoveries ranging between 640 and 1,710g/kg. The highest recoveries were found for polyunsaturated long-chain FA, the lowest for saturated or monounsaturated short- or medium-chain FA. A notable rearrangement of these FA components, particularly the minor ones, took place during ripening.

Short communication: Variations in major mineral contents of Mediterranean buffalo milk and application of Fourier-transform infrared spectroscopy for their prediction.

The aims of this study were (1) to assess variability in the major mineral components of buffalo milk, (2) to estimate the effect of certain environmental sources of variation on the major minerals during lactation, and (3) to investigate the possibility of using Fourier-transform infrared (FTIR) spectroscopy as an indirect, noninvasive tool for routine prediction of the mineral content of buffalo milk. A total of 173 buffaloes reared in 5 herds were sampled once during the morning milking. Milk samples were analyzed for Ca, P, K, and Mg contents within 3h of sample collection using inductively coupled plasma optical emission spectrometry. A Milkoscan FT2 (Foss, Hillerød, Denmark) was used to acquire milk spectra over the spectral range from 5,000 to 900 wavenumber/cm. Prediction models were built using a partial least square approach, and cross-validation was used to assess the prediction accuracy of FTIR. Prediction models were validated using a 4-fold random cross-validation, thus dividing the calibration-test set in 4 folds, using one of them to check the results (prediction models) and the remaining 3 to develop the calibration models. Buffalo milk minerals averaged 162, 117, 86, and 14.4mg/dL of milk for Ca, P, K, and Mg, respectively. Herd and days in milk were the most important sources of variation in the traits investigated. Parity slightly affected only Ca content. Coefficients of determination of cross-validation between the FTIR-predicted and the measured values were 0.71, 0.70, and 0.72 for Ca, Mg, and P, respectively, whereas prediction accuracy was lower for K (0.55). Our findings reveal FTIR to be an unsuitable tool when milk mineral content needs to be predicted with high accuracy. Predictions may play a role as indicator traits in selective breeding (if the additive genetic correlation between FTIR predictions and measures of milk minerals is high enough) or in monitoring the milk of buffalo populations for dairy industry purposes.

Genome-wide association of coagulation properties, curd firmness modeling, protein percentage, and acidity in milk from Brown Swiss cows.

Cheese production is increasing in many countries, and a desire toward genetic selection for milk coagulation properties in dairy cattle breeding exists. However, measurements of individual cheesemaking properties are hampered by high costs and labor, whereas traditional single-point milk coagulation properties (MCP) are sometimes criticized. Nevertheless, new modeling of the entire curd firmness and syneresis process (CFt equation) offers new insight into the cheesemaking process. Moreover, identification of genomic regions regulating milk cheesemaking properties might enhance direct selection of individuals in breeding programs based on cheese ability rather than related milk components. Therefore, the objective of this study was to perform genome-wide association studies to identify genomic regions linked to traditional MCP and new CFt parameters, milk acidity (pH), and milk protein percentage. Milk and DNA samples from 1,043 Italian Brown Swiss cows were used. Milk pH and 3 MCP traits were grouped together to represent the MCP set. Four CFt equation parameters, 2 derived traits, and protein percentage were considered as the second group of traits (CFt set). Animals were genotyped with the Illumina SNP50 BeadChip v.2 (Illumina Inc., San Diego, CA). Multitrait animal models were used to estimate variance components. For genome-wide association studies, the genome-wide association using mixed model and regression-genomic control approach was used. In total, 106 significant marker traits associations and 66 single nucleotide polymorphisms were identified on 12 chromosomes (1, 6, 9, 11, 13, 15, 16, 19, 20, 23, 26, and 28). Sharp peaks were detected at 84 to 88 Mbp on Bos taurus autosome (BTA) 6, with a peak at 87.4 Mbp in the region harboring the casein genes. Evidence of quantitative trait loci at 82.6 and 88.4 Mbp on the same chromosome was found. All chromosomes but BTA6, BTA11, and BTA28 were associated with only one trait. Only BTA6 was in common between MCP and CFt sets. The new CFt traits reinforced the support of MCP signals and provided with additional information on genomic regions that might be involved in regulation of the coagulation process of bovine milk.

Cheesemaking in highland pastures: Milk technological properties, cream, cheese and ricotta yields, milk nutrients recovery, and products composition.

Summer transhumance of dairy cows to high Alpine pastures is still practiced in many mountainous areas. It is important for many permanent dairy farms because the use of highland pastures increases milk production and high-priced typical local dairy products often boost farm income. As traditional cheese- and ricotta-making procedures in Alpine pastures are central to this dairy system, the objective of this study was to characterize the quality and efficiency of products and their relationships with the quality and availability of grass during the grazing season. The milk from 148 cows from 12 permanent farms reared on a temporary farm located in Alpine pastures was processed every 2wk during the summer (7 cheesemakings from late June to early September). During each processing, 11 dairy products (4 types of milk, 2 by-products, 3 fresh products, and 2 ripened cheeses) were sampled and analyzed. In addition, 8 samples of fresh forage from the pasture used by the cows were collected and analyzed. At the beginning of the pasture season the cows were at 233±90d in milk, 2.4±1.7 parities, and produced 23.6±5.7kg/d of milk. The milk yield decreased with the move from permanent to temporary farms and during the entire summer transhumance, but partly recovered after the cows returned to the permanent farms. Similar trends were observed for the daily yields of fat, protein, casein, lactose, and energy, as we found no large variations in the quality of the milk, with the exception of the first period of Alpine pasture. The somatic cell counts of milk increased during transhumance, but this resulted from a concentration of cells in a lower quantity of milk rather than an increase in the total number of cells ejected daily from the udder. We noted a quadratic trend in availability of forage (fresh and dry matter weight per hectare), with a maximum in late July. The quality of forage also varied during the summer with a worsening of chemical composition. The evening milk (before and after natural creaming), the whole morning milk, and the mixed vat milk had different chemical compositions, traditional coagulation properties, and curd-firming modeling parameters. These variations over the pasture season were similar to the residual variations with respect to chemical composition, and much lower with respect to coagulation and curd-firming traits. Much larger variations were noted in cream, cheese, and ricotta yields, as well as in nutrient recoveries in curd during the pasture season. The protein content of forage was correlated with some of the coagulation and curd-firming traits, the ether extract of forage was positively correlated with milk fat content and cheese yields, and fiber fractions of forage were unfavorably correlated with some of the chemical and technological traits. Traditional cheese- and ricotta-making procedures showed average cream, cheese, and ricotta yields of 6.3, 14.2, and 4.9%, respectively, and an overall recovery of almost 100% of milk fat, 88% of milk protein, and 60% of total milk solids.

Phenotypic analysis of cheese yields and nutrient recoveries in the curd of buffalo milk, as measured with an individual model cheese-manufacturing process.

Traits associated with cheese yield and milk nutrient recovery in curd are used to describe the efficiency of the cheese-making process. This is fundamental for all dairy species, including the Italian Mediterranean buffalo, which is largely used for milk production aimed at the dairy industry. To assess cheese-making traits among buffalo, a model cheese-manufacturing process was tested; it was capable of processing 24 samples per run, using 0.5-L samples of milk from individual buffalo. In total, 180 buffalo reared in 7 herds located in Northeast Italy were sampled once. Briefly, each sample was weighed and heated (35°C for 30min), inoculated with starter culture (90min), and mixed with rennet (51.2 international milk-clotting units/L of milk). After 10min of gelation, the curd was cut; 5min after the cut, the curd was separated from the whey, and the curd was subjected to draining (for 30min) and pressing (18h). The curd and whey were weighed, analyzed for pH and the total solid, fat, lactose, and protein contents, and subjected to estimation of the energy content. Three measures of cheese yield (%CY), %CYCURD, %CYSOLIDS, and %CYWATER, were computed as the ratios between the weight of the curd, the curd dry matter, and the water retained in the curd, respectively, and the weight of the milk processed. These traits were multiplied by the daily milk yield to define the 3 corresponding measures of daily cheese yield (dCY, kg/d). The milk component recoveries (REC) in the curd, RECFAT, RECPROTEIN, and RECSOLIDS, represented the ratios between the weights of the fat, protein, and total solids in the curd, respectively, and the corresponding components in the milk. Finally, energy recovery (RECENERGY) was estimated. The values for %CYCURD, %CYSOLIDS, %CYWATER, RECPROTEIN, RECFAT, RECSOLIDS, and RECENERGY averaged 25.6, 12.7, 12.9, 80.4, 95.1, 66.7, and 79.3%, respectively, indicating that buffalo milk has a higher aptitude to cheese-making than bovine milk. The effect of days in milk was the most important source of variation for %CY, RECPROTEIN, and the overall recoveries (which showed higher values toward the end of lactation), whereas parity did not appear to influence any of the investigated traits. The cheese-making procedure tested allowed us to assess the variability of and relationships among different cheese yield traits, recovery traits, daily milk production traits, and milk components at the individual level.

Genetic parameters of cheese yield and curd nutrient recovery or whey loss traits predicted using Fourier-transform infrared spectroscopy of samples collected during milk recording on Holstein, Brown Swiss, and Simmental dairy cows.

Cheese yield is the most important technological parameter in the dairy industry in many countries. The aim of this study was to infer (co)variance components for cheese yields (CY) and nutrient recoveries in curd (REC) predicted using Fourier-transform infrared (FTIR) spectroscopy of samples collected during milk recording on Holstein, Brown Swiss, and Simmental dairy cows. A total of 311,354 FTIR spectra representing the test-day records of 29,208 dairy cows (Holstein, Brown Swiss, and Simmental) from 654 herds, collected over a 3-yr period, were available for the study. The traits of interest for each cow consisted of 3 cheese yield traits (%CY: fresh curd, curd total solids, and curd water as a percent of the weight of the processed milk), 4 curd nutrient recovery traits (REC: fat, protein, total solids, and the energy of the curd as a percent of the same nutrient in the processed milk), and 3 daily cheese production traits (daily fresh curd, total solids, and the water of the curd per cow). Calibration equations (freely available upon request to the corresponding author) were used to predict individual test-day observations for these traits. The (co)variance components were estimated for the CY, REC, milk production, and milk composition traits via a set of 4-trait analyses within each breed. All analyses were performed using REML and linear animal models. The heritabilities of the %CY were always higher for Holstein and Brown Swiss cows (0.22 to 0.33) compared with Simmental cows (0.14 to 0.18). In general, the fresh cheese yield (%CYCURD) showed genetic variation and heritability estimates that were slightly higher than those of its components, %CYSOLIDS and %CYWATER. The parameter RECPROTEIN was the most heritable trait in all the 3 breeds, with values ranging from 0.32 to 0.41. Our estimation of the genetic relationships of the CY and REC with milk production and composition revealed that the current selection strategies used in dairy cattle are expected to exert only limited effects on the REC traits. Instead, breeders may be able to exploit genetic variations in the %CY, particularly RECFAT and RECPROTEIN. This last component is not explained by the milk protein content, suggesting that its direct selection could be beneficial for cheese production aptitude. Collectively, our findings indicate that breeding strategies aimed at enhancing CY and REC could be easily and rapidly implemented for dairy cattle populations in which FTIR spectra are routinely acquired from individual milk samples.

Milk skimming, heating, acidification, lysozyme, and rennet affect the pattern, repeatability, and predictability of milk coagulation properties and of curd-firming model parameters: A case study of Grana Padano.

Milk coagulation properties are used to evaluate the cheesemaking aptitude of milk samples. No international standard procedure exists, although laboratories often mimic the production of a full-fat fresh cheese for milk coagulation properties. Questions have arisen about the predictability of such a procedure for different types of cheese production. The aim of this study was to establish a procedure mimicking the production conditions of a long-ripened hard cheese, taking Protected Designation of Origin Grana Padano as a case study. With respect to the traditional conditions (standard procedure; SP), the Grana Padano procedure (GP) modifications were the use of standardized milk, coagulation lower temperature, previous milk acidification, lysozyme addition, and rennet type. Each modification was tested in turn versus the SP and also all together in the GP. Another 3 tests were carried out: SP on naturally creamed milk, SP with double the quantity of rennet, and a simplified GP on a full-fat milk sample. The 10 procedures were tested on 2 subsamples with 2 replicates each and were repeated using individual milk samples from 15 dual-purpose Simmental cows in 4 sessions for a total of 600 tests. Two Formagraph instruments (Foss Electric A/S, Hillerød, Denmark) measuring curd firmness every 15 s were used, prolonging test duration to 60min to obtain 5 traditional single-point milk coagulation properties and 3 parameters of the curd firming model using all 240 points recorded for each replicate. The 8 traits of each replicate were analyzed according to a mixed model with fixed effects of 4 sessions, 10 treatments, 2 instruments, and 16microvats, and random effects of 15 animals and 300 subsamples. Compared with the SP, the coagulation and curd firming was slowed by low temperature and was accelerated by acidification and by adding a double amount of rennet; natural creaming, fat standardization, and rennet with 5% pepsin affected only some traits, whereas lysozyme addition affected none. Combination of all modifications tended to compensate for each of their effects, resulting in similar average patterns between GP and SP. Modifications to repeatability were found for all traits with the exception of 2. The ability of the SP to predict GP, tested through correlations between procedures, was not very high. Whereas SP is used for both research and in the dairy industry, better results for Grana Padano cheesemaking can only be achieved by adopting specific, more complex, and labor-intensive procedures at the research level or, possibly, by specific calibrations through Fourier-transform infrared spectroscopy at the industry level.

Effect of dairy farming system, herd, season, parity, and days in milk on modeling of the coagulation, curd firming, and syneresis of bovine milk.

The objectives of this study were to characterize the variation in curd firmness model parameters obtained from coagulating bovine milk samples, and to investigate the effects of the dairy system, season, individual farm, and factors related to individual cows (days in milk and parity). Individual milk samples (n = 1,264) were collected during the evening milking of 85 farms representing different environments and farming systems in the northeastern Italian Alps. The dairy herds were classified into 4 farming system categories: traditional system with tied animals (29 herds), modern dairy systems with traditional feeding based on hay and compound feed (30 herds), modern dairy system with total mixed ration (TMR) that included silage as a large proportion of the diet (9 herds), and modern dairy system with silage-free TMR (17 herds). Milk samples were analyzed for milk composition and coagulation properties, and parameters were modeled using curd firmness measures (CFt) collected every 15 s from a lacto-dynamographic analysis of 90 min. When compared with traditional milk coagulation properties (MCP), the curd firming measures showed greater variability and yielded a more accurate description of the milk coagulation process: the model converged for 93.1% of the milk samples, allowing estimation of 4 CFt parameters and 2 derived traits [maximum CF (CF(max)) and time from rennet addition to CF(max) (t(max))] for each sample. The milk samples whose CFt equations did not converge showed longer rennet coagulation times obtained from the model (RCT(eq)) and higher somatic cell score, and came from less-productive cows. Among the sources of variation tested for the CFt parameters, dairy herd system yielded the greatest differences for the contrast between the traditional farm and the 3 modern farms, with the latter showing earlier coagulation and greater instant syneresis rate constant (k(SR)). The use of TMR yielded a greater tmax because of a higher instant curd-firming rate constant (k(CF)). Season of sampling was found to be very important, yielding higher values during winter for all traits except k(CF) and k(SR). All CFt traits were affected by individual cow factors. For parity, milk produced by first-lactation cows showed higher k(CF) and k(SR), but delays in achieving CF(max). With respect to stage of lactation, RCT(eq) and potential asymptotic CF increased during the middle of lactation and stabilized thereafter, whereas the 2 instant rate constants presented the opposite pattern, with the lowest (k(CF)) and highest (k(SR)) values occurring in mid lactation. The new challenge offered by prolonging the test interval and individual modeling of milk technological properties allowed us to study the effects of parameters related to the environment and to individual cows. This novel strategy may be useful for investigating the genetic variability of these new coagulation traits.

Effects of dairy system, herd within dairy system, and individual cow characteristics on the volatile organic compound profile of ripened model cheeses.

The objective of this work was to study the effect of dairy system, herd within dairy system, and characteristics of individual cows (parity, days in milk, and daily milk yield) on the volatile organic compound profile of model cheeses produced under controlled conditions from the milk of individual cows of the Brown Swiss breed. One hundred fifty model cheeses were selected from 1,272 produced for a wider study of the phenotypic and genetic variability of Brown Swiss cows. In our study, we selected 30 herds representing 5 different dairy systems. The cows sampled presented different milk yields (12.3-43.2kg/d), stages of lactation (10-412 d in milk), and parity (1-7). In total, 55 volatile compounds were detected by solid-phase microextraction and gas chromatography-mass spectrometry, including 14 alcohols, 13 esters, 11 free fatty acids, 8 ketones, 4 aldehydes, 3 lactones, 1 terpene, and 1 pyrazine. The most important sources of variation in the volatile organic profiles of model cheeses were dairy system (18 compounds) and days in milk (10 compounds), followed by parity (3 compounds) and milk yield (5 compounds). The model cheeses produced from the milk of tied cows reared on traditional farms had lower quantities of 3-methyl-butan-1-ol, 6-pentyloxan-2-one, 2-phenylethanol, and dihydrofuran-2(3H)-one compared with those reared in freestalls on modern farms. Of these, milk from farms using total mixed rations had higher contents of alcohols (hexan-1-ol, octan-1-ol) and esters (ethyl butanoate, ethyl pentanoate, ethyl hexanoate, and ethyl octanoate) and lower contents of acetic acid compared with those using separate feeds. Moreover, dairy systems that added silage to the total mixed ration produced cheeses with lower levels of volatile organic compounds, in particular alcohols (butan-1-ol, pentan-1-ol, heptan-1-ol), compared with those that did not. The amounts of butan-2-ol, butanoic acid, ethyl-2-methylpropanoate, ethyl-3-methylbutanoate, and 6-propyloxan-2-one increased linearly during lactation, whereas octan-1-ol, 3-methyl-3-buten-1-ol, 2-butoxyethanol, 6-pentyloxan-2-one, and 2,6-dimethylpyrazine showed a more complex pattern during lactation. The effect of the number of lactations (parity) was significant for octan-1-ol, butanoic acid, and heptanoic acid. Finally, concentrations of octan-1-ol, 2-phenylethanol, pentanoic acid, and heptanoic acid increased with increasing daily milk yield, whereas dihydrofuran-2(3H)-one decreased. In conclusion, the volatile organic compound profile of model cheeses from the milk of individual cows was affected by dairy farming system and stage of lactation and, to lesser extent, by parity and daily milk yield.

Proton transfer reaction time-of-flight mass spectrometry: A high-throughput and innovative method to study the influence of dairy system and cow characteristics on the volatile compound fingerprint of cheeses.

The aim of this work was to study the effect of dairy system and individual cow-related factors on the volatile fingerprint of a large number of individual model cheeses analyzed by proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS). A total of 1,075 model cheeses were produced using milk samples collected from individual Brown Swiss cows reared in 72 herds located in mountainous areas of Trento province (Italy). The herds belonged to 5 main dairy systems ranging from traditional to modern and the cows presented different daily milk yields (24.6±7.9kg × d(-1)), stages of lactation (199±138 d in milk), and parities (2.7±1.8). The PTR-ToF-MS revealed 619 peaks, of which the 240 most intense were analyzed, and 61 of these were tentatively attributed to relevant volatile organic compounds on the basis of their fragmentation patterns and data from the literature. Principal component analysis was used to convert the multiple responses characterizing the PTR-ToF-MS spectra into 5 synthetic variables representing 62% of the total information. These principal components were related to groups of volatile compounds tentatively attributed to different peaks and used to investigate the relationship of the volatile compound profile obtained by PTR-ToF-MS to animal and farm characteristics. Lactation stage is related to 4 principal components which brought together 52.9% of the total variance and 57.9% of the area of analyzed peaks. In particular, 2 principal components were positively related to peaks tentatively attributed to aldehydes and ketones and negatively related to alcohols, esters, and acids, which displayed a linear increase during lactation. The second principal component was affected by dairy system; it was higher in the modern system in which cows received total mixed rations. The third principal component was positively related to daily milk production. In summary, we report the first application of this innovative, high-throughput technique to study the effects of dairy system and individual animal factors on volatile organic compounds of model cheeses. Individual cheesemaking procedures together with this spectrometric technique open new avenues for genetic selection of dairy species with respect to both milk and cheese quality.

Modeling of coagulation, curd firming, and syneresis of milk from Sarda ewes.

This study investigated the modeling of curd-firming (CF) over time (CF(t)) of sheep milk. Milk samples from 1,121 Sarda ewes from 23 flocks were analyzed for coagulation properties. Lactodynamographic analyses were conducted for up to 60 min, and 240 CF individual observations from each sample were recorded. Individual sample CFt equation parameters (RCT(eq), rennet coagulation time; CF(P), asymptotic potential value of curd firmness; k(CF), curd-firming instant rate constant; and k(SR), curd syneresis instant rate constant) were estimated, and the derived traits (CF(max), the point at which CF(t) attained its maximum level, and tmax, the time at which CF(max) was attained) were calculated. The incidence of noncoagulating milk samples was 0.4%. The iterative estimation procedure applied to the individual coagulation data showed a small number of not-converged samples (4.4%), which had late coagulation and an almost linear pattern of the ascending part of the CF(t) curve that caused a high value of CF(P), a low value of k(CF), and a high value of k(SR). Converged samples were classified on the basis of their CF(t) curves into no-k(SR) (18.0%), low-k(SR) (72.6%), and high-k(SR) (4.5%). A CF(t) that was growing continuously because of the lack of the syneresis process characterized the no-k(SR) samples. The high-k(SR) samples had a much larger CFP, a smaller k(CF), and an anticipation of tmax, whereas the low-k(SR) samples had a fast k(CF) and a slower k(SR). The part of the average CF(t) curves that showed an increase was similar among the 3 different syneretic groups, whereas the part that decreased was different because of the expulsion of whey from the curd. The traditional milk coagulation properties recorded within 30 min were not able to detect any appreciable differences among the 4 groups of coagulating samples, which could lead to a large underestimation of the maximum CF of all samples (if predicted by a30), with the exception of the no-k(SR) samples. Large individual variability was found and was likely caused by the effects of the dairy system, such as flock size (on CF(max), t(max), and % ewes with no-k(SR) milk), flock within flock size (representing 11 to 43% of total variance for % ewes with no-k(SR) milk and CF(max), respectively), days in milk (on all model parameters and CF(max)), parity (on RCT(eq), k(SR), and CF(max)), daily milk yield (on RCT(eq) and CF(max)), and position of the individual pendulum that significantly affected model parameters and derived traits. In conclusion, the results showed that the modeling of coagulation, curd-firming, and syneresis is a suitable tool to achieve a deeper interpretation of the coagulation and curd-firming processes of sheep milk and also to study curd syneresis.

Comparison between genetic parameters of cheese yield and nutrient recovery or whey loss traits measured from individual model cheese-making methods or predicted from unprocessed bovine milk samples using Fourier-transform infrared spectroscopy.

Cheese yield is an important technological trait in the dairy industry. The aim of this study was to infer the genetic parameters of some cheese yield-related traits predicted using Fourier-transform infrared (FTIR) spectral analysis and compare the results with those obtained using an individual model cheese-producing procedure. A total of 1,264 model cheeses were produced using 1,500-mL milk samples collected from individual Brown Swiss cows, and individual measurements were taken for 10 traits: 3 cheese yield traits (fresh curd, curd total solids, and curd water as a percent of the weight of the processed milk), 4 milk nutrient recovery traits (fat, protein, total solids, and energy of the curd as a percent of the same nutrient in the processed milk), and 3 daily cheese production traits per cow (fresh curd, total solids, and water weight of the curd). Each unprocessed milk sample was analyzed using a MilkoScan FT6000 (Foss, Hillerød, Denmark) over the spectral range, from 5,000 to 900 wavenumber × cm(-1). The FTIR spectrum-based prediction models for the previously mentioned traits were developed using modified partial least-square regression. Cross-validation of the whole data set yielded coefficients of determination between the predicted and measured values in cross-validation of 0.65 to 0.95 for all traits, except for the recovery of fat (0.41). A 3-fold external validation was also used, in which the available data were partitioned into 2 subsets: a training set (one-third of the herds) and a testing set (two-thirds). The training set was used to develop calibration equations, whereas the testing subsets were used for external validation of the calibration equations and to estimate the heritabilities and genetic correlations of the measured and FTIR-predicted phenotypes. The coefficients of determination between the predicted and measured values in cross-validation results obtained from the training sets were very similar to those obtained from the whole data set, but the coefficient of determination of validation values for the external validation sets were much lower for all traits (0.30 to 0.73), and particularly for fat recovery (0.05 to 0.18), for the training sets compared with the full data set. For each testing subset, the (co)variance components for the measured and FTIR-predicted phenotypes were estimated using bivariate Bayesian analyses and linear models. The intraherd heritabilities for the predicted traits obtained from our internal cross-validation using the whole data set ranged from 0.085 for daily yield of curd solids to 0.576 for protein recovery, and were similar to those obtained from the measured traits (0.079 to 0.586, respectively). The heritabilities estimated from the testing data set used for external validation were more variable but similar (on average) to the corresponding values obtained from the whole data set. Moreover, the genetic correlations between the predicted and measured traits were high in general (0.791 to 0.996), and they were always higher than the corresponding phenotypic correlations (0.383 to 0.995), especially for the external validation subset. In conclusion, we herein report that application of the cross-validation technique to the whole data set tended to overestimate the predictive ability of FTIR spectra, give more precise phenotypic predictions than the calibrations obtained using smaller data sets, and yield genetic correlations similar to those obtained from the measured traits. Collectively, our findings indicate that FTIR predictions have the potential to be used as indicator traits for the rapid and inexpensive selection of dairy populations for improvement of cheese yield, milk nutrient recovery in curd, and daily cheese production per cow.

Milk quality, coagulation properties, and curd firmness modeling of purebred Holsteins and first- and second-generation crossbred cows from Swedish Red, Montbéliarde, and Brown Swiss bulls.

The objective of the present study was to investigate how the crossbreeding of Holstein (HO) cows with bulls from Nordic and Alpine European breeds affect milk quality traits, traditional milk coagulation properties (MCP), and curd firmness modeling obtained from individual milk samples. A total of 506 individual milk samples were collected from evening milking at 3 commercial farms located in Northern Italy. Over the past decade, the 3 farms have followed crossbreeding programs in part of their herds, whereas the remainder of the animals consisted of purebred HO. The basic scheme was a 3-breed rotation based on the use of Swedish Red (SR) semen on HO cows (SR × HO), the use of Montbéliarde (MO) semen on first-cross cows [MO × (SR × HO)], and the use of HO semen in the third cross. In all herds, a smaller proportion of purebred HO were mated to M and Brown Swiss (BS) bulls, and these first crosses were mated to SR and MO bulls, respectively. Milk samples were analyzed for milk composition and MCP, and parameters for curd firmness were modeled. Compared with purebred HO, crossbred cows produced less milk with lower lactose content, higher fat and protein content, and a tendency for higher casein content. Crossbred cows generally produced milk with a more favorable curd-firming rate (k20) and curd firmness 30 min after rennet addition, among traditional MCP, and better trends of curd firmness measures as shown by model parameters: estimated rennet coagulation time, asymptotical potential value of curd firmness, and curd-firming instant rate constant. Among crossbred cows, SR × HO presented longer rennet coagulation time compared with MO × HO and BS × HO cows, and MO × HO showed shorter k20 compared with BS × HO cows. Among second-generation cows, those sired by SR bulls showed a lower incidence of noncoagulated samples, higher curd firmness 30 min after rennet addition and asymptotical potential value of curd firmness, and faster curd-firming instant rate constant compared with animals sired by MO bulls. Our results revealed that different sire breeds were characterized by specific technological aptitudes, but that these were not strictly related to other milk quality traits. Furthermore, the favorable characteristics (in terms of the quality and technological properties of milk) could be maintained in the third generation of 3-way crosses without negative effects on milk yield, even though the HO heritage had been reduced from 50 to 25%. Our findings, therefore, suggest that different types of sires can be chosen (depending on the intended use of the milk) to ensure the optimization of farm crossbreeding programs.