The use of milk Fourier-transform infrared spectra for predicting cheesemaking traits in Grana Padano Protected Designation of Origin cheese.

The prediction of the cheese yield (%CY) traits for curd, solids, and retained water and the amount of fat, protein, solids, and energy recovered from the milk into the curd (%REC) by Bayesian models, using Fourier-transform infrared spectroscopy (FTIR), can be of significant economic interest to the dairy industry and can contribute to the improvement of the cheese process efficiency. The yields give a quantitative measure of the ratio between weights of the input and output of the process, whereas the nutrient recovery allows to assess the quantitative transfer of a component from milk to cheese (expressed in % of the initial weight). The aims of this study were: (1) to investigate the feasibility of using bulk milk spectra to predict %CY and %REC traits, and (2) to quantify the effect of the dairy industry and the contribution of single-spectrum wavelengths on the prediction accuracy of these traits using vat milk samples destined to the production of Grana Padano Protected Designation of Origin cheese. Information from 72 cheesemaking days (in total, 216 vats) from 3 dairy industries were collected. For each vat, the milk was weighed and analyzed for composition (total solids [TS], lactose, protein, and fat). After 48 h from cheesemaking, each cheese was weighed, and the resulting whey was sampled for composition as well (TS, lactose, protein, and fat). Two spectra from each milk sample were collected in the range between 5,011 and 925 cm-1 and averaged before the data analysis. The calibration models were developed via a Bayesian approach by using the BGLR (Bayesian Generalized Linear Regression) package of R software. The performance of the models was assessed by the coefficient of determination (R2VAL) and the root mean squared error (RMSEVAL) of validation. Random cross-validation (CVL) was applied [80% calibration and 20% validation set] with 10 replicates. Then, a stratified cross-validation (SCV) was performed to assess the effect of the dairy industry on prediction accuracy. The study was repeated using a selection of informative wavelengths to assess the necessity of using whole spectra to optimize prediction accuracy. Results showed the feasibility of using FTIR spectra and Bayesian models to predict cheesemaking traits. The R2VAL values obtained with the CVL procedure were promising in particular for the %CY and %REC for protein, ranging from 0.44 to 0.66 with very low RMSEVAL (from 0.16 to 0.53). Prediction accuracy obtained with the SCV was strongly influenced by the dairy factory industry. The general low values gained with the SCV do not permit a practical application of this approach, but they highlight the importance of building calibration models with a dataset covering the largest possible sample variability. This study also demonstrated that the use of the full FTIR spectra may be redundant for the prediction of the cheesemaking traits and that a specific selection of the most informative wavelengths led to improved prediction accuracy. This could lead to the development of dedicated spectrometers using selected wavelengths with built-in calibrations for the online prediction of these innovative traits.

Genomic inbreeding coefficients using imputation genotypes: assessing the effect of ancestral genotyping in Holstein-Friesian dairy cows.

The objective of this study was to assess the effect of using or not the genotypes of the parents of a cow for imputing single nucleotide polymorphisms (SNP), on the estimation of genomic inbreeding coefficients of cows. Imputation (i.e., genotyped plus imputed) genotypes from 68,127 Italian Holstein dairy cows registered in the Italian National Association of Holstein, Brown and Jersey Breeders (ANAFIBJ) were analyzed. Cows were genotyped with the HD Illumina Infinium BovineHD BeadChip and GeneSeek Genomic Profiler HD-150K, and the MD GeneSeek Genomic Profiler 3, GeneSeek Genomic Profiler 4, GeneSeek MD and the Labogena MD. To assess differences among estimators genomic inbreeding coefficients were estimated with 4 PLINK v1.9 estimators (F, Fhat1, 2, 3), 2 genomic relationship matrix (grm) based estimators (Fgrm and Fgrm2; with the latter including also pedigree information) and one estimator of runs of homozygosity (ROH; FROH). Assuming that the correct genomic inbreeding coefficients should be those estimated from genotyped SNP, a comparison of the genomic inbreeding coefficients estimated either with the genotyped SNP or the SNP after imputation was made. Information on the presence or absence of genotypic information from sire, dam and maternal grandsire during the imputation was investigated. Genomic inbreeding coefficients estimated with genotyped SNP or SNP after imputation were consistent for F, Fhat3, Fgrm2 and FROH, when at least one of the parents was genotyped. Biased (mainly higher) genomic inbreeding coefficients of imputation SNP were observed in cows that were genotyped with MD SNP panels whose SNP were poorly represented in the selected imputation SNP data set and also did not have their parents genotyped compared with what expected based on actual genotype data. For cows genotyped with MD the estimators Fhat1, Fhat2 and Fgrm provided higher genomic inbreeding coefficients of imputation SNP even with both parents and the maternal grandsire genotyped. Overall, FROH was the most robust estimator, followed by F and Fhat3. Our findings suggest that SNP selection, parental genotyping and estimator should be considered for designing imputation strategies in dairy cattle for estimating genomic inbreeding with imputation SNP. For computing genomic inbreeding coefficients, it is recommendable to have at least one parent genotyped and use an ROH based estimator.

Microsatellite imputation using SNP data for parentage verification in four Italian sheep breeds.

Microsatellite markers (MS) have been widely used for parentage verification in most of the livestock species over the past decades mainly due to their high polymorphic information content. In the genomic era, the spread of genotype information as single-nucleotide polymorphism (SNP) has raised the question to effectively use SNPs also for parentage testing. Despite the clear advantages of SNP panels in terms of cost, accuracy, and automation, the transition from MS to SNP markers for parentage verification is still very slow and, so far, only routinely applied in cattle. A major difficulty during this transition period is the need of SNP data for parents and offspring, which in most cases is not yet feasible due to the genotyping cost. To overcome the unavailability of same genotyping platform during the transition period, in this study we aimed to assess the feasibility of a MS imputation pipeline from SNPs in four native sheep dairy breeds: Comisana (N = 331), Massese (N = 210), Delle Langhe (N = 59) and Sarda (N = 1003). Those sheep were genotyped for 11 MS and with the Ovine SNP50 Bead Chip. Prior to imputation, a quality control (QC) was performed, and SNPs located within a window of 2 Mb from each MS were selected. The core of the developed pipeline was made up of three steps: (a) storing of both MS and SNP data in a Variant Call Format file, (b) masking MS information in a random sample of individuals (10%), (c) imputing masked MS based on non-missing individuals (90%) using an imputation program. The feasability of the proposed methodology was assessed also among different training - testing split ratio, population size, number of flanking SNPs as well as within and among breeds. The accuracy of the MS imputation was assessed based on the genotype concordance as well as at parentage verification level in a subset of animals in which assigned parents' MS were available. A total of 8 MS passed the QC, and 505 SNPs were located within the ±2 Mb window from each MS, with an average of 63 SNPs per MS. The results were encouraging since when excluding the worst imputed MS (OARAE129), and regardless on the analyses performed (within and across breeds) for all breeds, we achieved an overall concordance rate over 94%. In addition, on average, the imputed offspring MS resulted in equivalent parentage outcome in 94% of the cases when compared to verification using original MS, highlighting both the feasibility and the eventual practical advantage of using this imputation pipeline.

Heritability and genetic correlations of total and differential somatic cell count with milk yield and composition traits in Italian Simmental cows.

Costs of production have deeply increased each year in the last decades, breeders are continuously looking for more cost effective and more efficient ways to produce milk. Despite the major signs of progress in productivity, it is fundamental to optimize rather than maximize the performances of the dairy cows. Mastitis is still a highly prevalent disease in the dairy sector which causes several economic losses and environmental effect. Its accurate and early diagnosis is crucial to improve profitability of dairy cows and contribute to a more sustainable dairy industry. Among mastitis reduction strategies, there is the urgent need to implement breeding objectives to select cows displaying mastitis resistance by investigating the genetic mechanisms at the base of the inflammatory response. Therefore, in this study we aimed to further understand the genetic background of the differential somatic cell count (DSCC), which provides thorough insights on the actual inflammatory status of the mammary glands. The objectives of this study were to estimate on a cohort of 20,215 Italian Simmental cows over a 3-yr period: (1) the heritability and repeatability values of somatic cell score (SCS) and DSCC, (2) the genetic and phenotypic correlations between these 2 traits and milk production and milk composition traits, (3) the heritability and repeatability values of SCS and DSCC within class of udder health status. Heritability was low both for SCS (0.06) and DSCC (0.08), whereas the repeatability values for these traits were 0.43 and 0.36, suggesting that the magnitude of cow permanent environmental effect for these traits is remarkable. The genetic and phenotypic correlation of SCS with DSCC was 0.612 and 0.605, respectively. Because both significantly differed from the unit, we must consider those traits as different ones. This latter aspect corroborates the need to consider the DSCC as a further indicator of inflammatory status which might be implemented in the Simmental breed genetic evaluation. It is worthy to mention that heritability estimates for SCS and DSCC were the highest in healthy cows compared with the other udder health classes. This implies that when the udder health status changes, it is most likely due to environmental factors rather than aspects related to the animal's genetics. In contrast, the highest additive genetic variance and heritability found for SCS and DSCC in the healthy group might reveal the potential to further implement breeding strategies to select for healthier animals.

Cheese yield and nutrients recovery in the curd predicted by Fourier-transform spectra from individual sheep milk samples.

The objectives of this study were to explore the use of Fourier-transform infrared (FTIR) spectroscopy on individual sheep milk samples for predicting cheese-making traits, and to test the effect of the farm variability on their prediction accuracy. For each of 121 ewes from 4 farms, a laboratory model cheese was produced, and 3 actual cheese yield traits (fresh cheese, cheese solids, and cheese water) and 4 milk nutrient recovery traits (fat, protein, total solids, and energy) in the curd were measured. Calibration equations were developed using a Bayesian approach with 2 different scenarios: (1) a random cross-validation (80% calibration; 20% validation set), and (2) a leave-one-out validation (3 farms used as calibration, and the remaining one as validation set) to assess the accuracy of prediction of samples from external farms, not included in calibration set. The best performance was obtained for predicting the yield and recovery of total solids, justifying for the practical application of the method at sheep population and dairy industry levels. Performances for the remaining traits were lower, but still useful for the monitoring of the milk processing in the case of fresh curd and recovery of energy. Insufficient accuracies were found for the recovery of protein and fat, highlighting the complex nature of the relationships among the milk nutrients and their recovery in the curd. The leave-one-out validation procedure, as expected, showed lower prediction accuracies, as a result of the characteristics of the farming systems, which were different between calibration and validation sets. In this regard, the inclusion of information related to the farm could help to improve the prediction accuracy of these traits. Overall, a large contribution to the prediction of the cheese-making traits came from the areas known as "water" and "fingerprint" regions. These findings suggest that, according to the traits studied, the inclusion of water regions for the development of the prediction equation models is fundamental to maintain a high prediction accuracy. However, further studies are necessary to better understand the role of specific absorbance peaks and their contribution to the prediction of cheese-making traits, to offer reliable tools applicable along the dairy ovine chain.

Predictive formulas for different measures of cheese yield using milk composition from individual goat samples.

The objective of this study was to develop formulas based on milk composition of individual goat samples for predicting cheese yield (%CY) traits (fresh curd, milk solids, and water retained in the curd). The specific aims were to assess and quantify (1) the contribution of major milk components (fat, protein, and casein) and udder health indicators (lactose, somatic cell count, pH, and bacterial count) on %CY traits (fresh curd, milk solids, and water retained in the curd); (2) the cheese-making method; and (3) goat breed effects on prediction accuracy of the %CY formulas. The %CY traits were analyzed in duplicate from 600 goats, using an individual laboratory cheese-making procedure (9-MilCA method; 9 mL of milk per observation) for a total of 1,200 observations. Goats were reared in 36 herds and belonged to 6 breeds (Saanen, Murciano-Granadina, Camosciata delle Alpi, Maltese, Sarda, and Sarda Primitiva). Fresh %CY (%CYCURD), total solids (%CYSOLIDS), and water retained (%CYWATER) in the curd were used as response variables. Single and multiple linear regression models were tested via different combinations of standard milk components (fat, protein, casein) and indirect udder health indicators (UHI; lactose, somatic cell count, pH, and bacterial count). The 2 %CY observations within animal were averaged, and a cross-validation (CrV) scheme was adopted, in which 80% of observations were randomly assigned to the calibration (CAL) set and 20% to the validation (VAL) set. The procedure was repeated 10 times to account for sampling variability. Further, the model presenting the best prediction accuracy in CrV (i.e., comprehensive formula) was used in a secondary analysis to assess the accuracy of the %CY predictive formulas as part of the laboratory cheese-making procedure (within-animal validation, WAV), in which the first %CY observation within animal was assigned to CAL, and the second to the VAL set. Finally, a stratified CrV (SCrV) was adopted to assess the %CY traits prediction accuracy across goat breeds, again using the best model, in which 5 breeds were included in CAL and the remaining one in the VAL set. Fitting statistics of the formulas were assessed by coefficient of determination of validation (R2VAL) and the root mean square error of validation (RMSEVAL). In CrV, the formula with the best prediction accuracy for all %CY traits included fat, casein, and UHI (R2VAL = 0.65, 0.96, and 0.23 for %CYCURD, %CYSOLIDS, and %CYWATER, respectively). The WAV procedure showed R2VAL higher than those obtained in CrV, evidencing a low effect of the 9-MilCA method and, indirectly, its high repeatability. In the SCrV, large differences for %CYCURD and %CYWATER among breeds evidenced that the breed is a fundamental factor to consider in %CY predictive formulas. These results may be useful to monitor milk composition and quantify the influence of milk traits in the composite selection indices of specific breeds, and for the direct genetic improvement of cheese production.

Genomic inbreeding coefficients using imputed genotypes: Assessing different estimators in Holstein-Friesian dairy cows.

The objective of this study was to estimate inbreeding coefficients in Holstein dairy cattle using imputed SNPs data. A data set of 95,540 Italian Holstein dairy cows from the routine genomic evaluations of the Italian National Association of Holstein, Brown, and Jersey Breeders were analyzed, with 84,445 imputed SNP. Ten widely used genomic inbreeding estimators were tested, including 4 PLINK v1.9 estimators (F, FHAT1, FHAT2, FHAT3), 3 genomic relationship matrix (GRM)-based methods [VanRaden's first method with observed allele frequencies (FGRM) or with fixed frequencies at 0.5 (FGRM05), VanRaden's third method, allelic frequency free and pedigree regressed (FGRM2)], runs of homozygosity (ROH)-based estimators in a complete (FROH) and simplified version (FROH2), and proportion of homozygous SNP (FPH). Pairwise comparisons among them were made, including the comparison with traditional pedigree-based inbreeding coefficients (FPED). Our results showed variability among the genomic inbreeding estimators. Coefficients of FGRM and FHAT3 were >1, meaning that more variability has been lost than the variability that existed in the base population. Regarding the remaining ones, FGRM05, FROH, FROH2, and FPH provided coefficients within the [0,1] space and are considered comparable to FPED. Not comparable to FPED, yet with an interpretable value, can be considered the coefficients of F, FHAT2, and FGRM2. Estimators based on ROH had the highest correlation with pedigree-based coefficients (0.59-0.66), among all estimators tested. In this study, Spearman correlations were shown to possibly provide a clearer estimation of the strength of the relationship between estimators. We hypothesize that imputation might cause extreme genomic inbreeding values that deserves further investigation.

The mineral profile affects the coagulation pattern and cheese-making efficiency of bovine milk.

Natural variations in milk minerals, their relationships, and their associations with the coagulation process and cheese-making traits present an opportunity for the differentiation of milk destined for high-quality natural products, such as traditional specialties or Protected Designation of Origin (PDO) cheeses. The aim of this study was to quantify the effects of the native contents of Ca, P, Na, K, and Mg on 18 traits describing traditional milk coagulation properties (MCP), curd firming over time (CFt) equation parameters, cheese yield (CY) measures, and nutrient recoveries in the curd (REC) using models that either included or omitted the simultaneous effects of milk fat and casein contents. The results showed that, by including milk fat and casein and the minerals in the statistical model, we were able to determine the specific effects of each mineral on coagulation and cheese-making efficiency. In general, about two-thirds of the apparent effects of the minerals on MCP and the CFt equation parameters are actually mediated by their association with milk composition, especially casein content, whereas only one-third of the effects are direct and independent of milk composition. In the case of cheese-making traits, the effects of the minerals were mediated only negligibly by their association with milk composition. High Ca content had a positive effect on the coagulation pattern and cheese-making traits, favoring water retention in the curd in particular. Phosphorus positively affected the cheese-making traits in that it was associated with an increase in CY in terms of curd solids, and in all the nutrient recovery traits. However, a very high P content in milk was associated with lower fat recovery in the curd. The variation in the Na content in milk only mildly affected coagulation, whereas with regard to cheese-making, protein recovery was negatively associated with high concentrations of this mineral. Potassium seemed not to be actively involved in coagulation and the cheese-making process. Magnesium content tended to slow coagulation and reduce CY measures. Further studies on the relationships of minerals with casein and protein fractions could deepen our knowledge of the role of all minerals in coagulation and the cheese-making process.

Breed of goat affects the prediction accuracy of milk coagulation properties using Fourier-transform infrared spectroscopy.

The prediction of traditional goat milk coagulation properties (MCP) and curd firmness over time (CFt) parameters via Fourier-transform infrared (FTIR) spectroscopy can be of significant economic interest to the dairy industry and can contribute to the breeding objectives for the genetic improvement of dairy goat breeds. Therefore, the aims of this study were to (1) explore the variability of milk FTIR spectra from 4 goat breeds (Camosciata delle Alpi, Murciano-Granadina, Maltese, and Sarda), and to assess the possible discriminant power of milk FTIR spectra among breeds, (2) assess the viability to predict coagulation traits by using milk FTIR spectra, and (3) quantify the effect of the breed on the prediction accuracy of MCP and CFt parameters. In total, 611 individual goat milk samples were used. Analysis of variance of measured MCP and CFt parameters was carried out using a mixed model including the farm and pendulum as random factors, and breed, parity, and days in milk as fixed factors. Milk spectra for each goat were collected over the spectral range from wavenumber 5,011 to 925 × cm-1. Discriminant analysis of principal components was used to assess the ability of FTIR spectra to identify breed of origin. A Bayesian model was used to calibrate equations for each coagulation trait. The accuracy of the model and the prediction equation was assessed by cross-validation (CRV; 80% training and 20% testing set) and stratified CRV (SCV; 3 breeds in the training set, one breed in the testing set) procedures. Prediction accuracy was assessed by using coefficient of determination of validation (R2VAL), the root mean square error of validation (RMSEVAL), and the ratio performance deviation. Moreover, measured and FTIR predicted traits were compared in the SCV procedure by assessing their least squares means for the breed effect, Pearson correlations, and variance heteroscedasticity. Results showed the feasibility of using FTIR spectra and multivariate analyses to correctly assign milk samples to their breeds of origin. The R2VAL values obtained with the CRV procedure were moderate to high for the majority of coagulation traits, with RMSEVAL and ratio performance deviation values increasing as the coagulation process progresses from rennet addition. Prediction accuracy obtained with the SCV were strongly influenced by the breed, presenting general low values restricting a practical application. In addition, the low Pearson correlation coefficients of Sarda breed for all the traits analyzed, and the heteroscedastic variances of Camosciata delle Alpi, Murciano-Granadina, and Maltese breeds, further indicated that it is fundamental to consider the differences existing among breeds for the prediction of milk coagulation traits.

Goat farm variability affects milk Fourier-transform infrared spectra used for predicting coagulation properties.

Driven by the large amount of goat milk destined for cheese production, and to pioneer the goat cheese industry, the objective of this study was to assess the effect of farm in predicting goat milk-coagulation and curd-firmness traits via Fourier-transform infrared spectroscopy. Spectra from 452 Sarda goats belonging to 14 farms in central and southeast Sardinia (Italy) were collected. A Bayesian linear regression model was used, estimating all spectral wavelengths' effects simultaneously. Three traditional milk-coagulation properties [rennet coagulation time (min), time to curd firmness of 20 mm (min), and curd firmness 30 min after rennet addition (mm)] and 3 curd-firmness measures modeled over time [rennet coagulation time estimated according to curd firmness change over time (RCTeq), instant curd-firming rate constant, and asymptotical curd firmness] were considered. A stratified cross validation (SCV) was assigned, evaluating each farm separately (validation set; VAL) and keeping the remaining farms to train (calibration set) the statistical model. Moreover, a SCV, where 20% of the goats randomly taken (10 replicates per farm) from the VAL farm entered the calibration set, was also considered (SCV80). To assess model performance, coefficient of determination (R2VAL) and the root mean squared error of validation were recorded. The R2VAL varied between 0.14 and 0.45 (instant curd-firming rate constant and RCTeq, respectively), albeit the standard deviation was approximating half of the mean for all the traits. Although average results of the 2 SCV procedures were similar, in SCV80, the maximum R2VAL increased at about 15% across traits, with the highest observed for time to curd firmness of 20 mm (20%) and the lowest for RCTeq (6%). Further investigation evidenced important variability among farms, with R2VAL for some of them being close to 0. Our work outlined the importance of considering the effect of farm when developing Fourier-transform infrared spectroscopy prediction equations for coagulation and curd-firmness traits in goats.

Short communication: Dietary protein restriction and conjugated linoleic acid supplementation in dairy cows affect milk composition, the cheese-making process, and cheese quality.

We used 20 mid-lactating Holstein cows, housed in 4 pens according to a Latin square design, to evaluate the effects of dietary protein restriction (crude protein: 12.3 vs. 15.0% dry matter) and conjugated linoleic acid supplementation (CLA: 6.34 g/d of C18:2cis-9,trans-11 and 6.14 g/d of C18:2trans-10,cis-12) on milk composition, coagulation, curd firming and syneresis modeling, and cheese yield and quality (96 small cheeses). Dietary crude protein restriction, suggested as a way to reduce N excretion in farming, caused a reduction in milk protein content (-4%,), milk casein (-3.8%), lactose (-1%), cheese soluble protein (-16.8%), and the cheese maturation index (-15%), and a correlated increase in cheese fat content (+7.5%) and the fat to protein ratio (+18%). A modest reduction (-0.9%) in milk fat recovery in the curd did not affect cheese yield. The addition of CLA to the cows' diet, suggested as a way to improve N use efficiency and the nutritional value of dairy products, caused substantial alterations to the milk composition, cheese-making process, and cheese quality. The CLA reduced the fat (-12.3%), protein (-2%), casein (-2.2%), lactose (-1.0), and total solids (-4%) contents of milk, tended to delay coagulation, and weakened curd firming. The CLA reduced the fresh cheese yield (-7.5%) and cheese solids (-8.2%) because of the lower nutrient content of the milk, but also because of a lower recovery of milk protein in the curd (-0.9%) and lower total solids (-4.5%). It also reduced the fat content in the ripened cheese (-11.8%), as well as the fat to protein ratio (-19.4%) as a result of having increased the protein content (+9.3%). Last, it increased the lightness of the paste of the ripened cheeses (+3.3%), and especially the shear force (+16.3%). Dietary crude protein restriction, and CLA addition in particular, substantially altered the milk composition, cheese-making process, and cheese quality, and therefore needs to be carefully evaluated. Further studies are required to shed light on the causes of these modifications.

Modeling weight loss of cheese during ripening and the influence of dairy system, parity, stage of lactation, and composition of processed milk.

The yield, flavor, and texture of ripened cheese result from numerous interrelated microbiological, biochemical, and physical reactions that take place during ripening. The aims of the present study were to propose a 2-compartment first-order kinetic model of cheese weight loss over the ripening period; to test the variation in new informative phenotypes describing this process; and to assess the effects on these traits of dairy farming system, individual farms within dairy system, animal factors, and milk composition. A total of 1,211 model cheeses were produced in the laboratory using individual 1.5-L milk samples from Brown Swiss cows reared on 83 farms located in Trento Province. During ripening (60 d; temperature 15°C, relative humidity 85%), the weight of all model cheeses was measured, and cheese yield (cheese weight/processed milk weight, %CY) was calculated at 7 intervals from cheese-making (0, 1, 7, 14, 28, 42, and 60 d). Using these measures, a 2-compartment first-order kinetic model (3-parameter equation) was developed for modeling %CY during the ripening period, as follows: [Formula: see text] , where %CYt is the %CY at ripening time t; %CYi and %CYf are the modeled %CY traits at time 0 d (%CYi = initial %CY) and at the end of a ripening period sufficient to reach a constant wheel weight (%CYf = final %CY after 60 d ripening in the case of small model cheeses); kCY is the instant rate constant for cheese weight loss (%/d). Cheese weight and protein and fat losses were calculated as the % difference between the model cheeses at 0 and after 60 d of ripening. The variation in cheese pH was calculated as the % difference between pH at 0 and after 60 d. Dairy system, individual herd within dairy system, and the cow's parity and lactation stage (tested with a linear mixed model) strongly affected almost all the traits collected during model cheese ripening. Milk fat, protein, lactose, pH, and somatic cell score also greatly affected almost all the traits, although kCY was affected only by milk protein. After including milk composition in the linear mixed model, the importance of all the herd and animal sources of variation was greatly reduced for all traits. The proposed model and novel traits could be tested, first, with the aim of establishing new monitoring procedures enabling the dairy industry to improve milk quality-based payment systems at the herd level and, second, with a view to exploring possible genetic improvements to dairy cow populations.

Characterization of milk composition, coagulation properties, and cheese-making ability of goats reared in extensive farms.

The aims of this study were to explore the variability of milk composition, coagulation properties, and cheese-making traits of the Sarda goat breed, and to investigate the effects of animal and farm factors, and the geographic area (Central-East vs. South-West) of an insular region of Italy, Sardinia. A total of 570 Sarda goats reared in 21 farms were milk-sampled during morning milking. Individual milk samples were analyzed for composition, traditional milk coagulation properties (MCP), modeled curd-firming over time parameters (CFt), and cheese-making traits (cheese yield, %CY; recovery of nutrients, %REC; daily cheese yield, dCY). Farms were classified into 2 categories based on milk energy level (MEL; high or low), defined according to the average net energy of milk daily produced by the lactating goats. Milk yield and composition were analyzed using a mixed model including the fixed effects of MEL, geographic area, days in milk, and parity, and the random effect of farm within MEL and geographic area. Data about MCP, CFt, and the cheese-making process were analyzed using the same model, with the inclusion of the effects of animal and pendulum of the lactodynamograph instrument, allowing the measure of repeatability of these traits. Results showed that animal had greater influence on coagulation and cheese-making traits compared with farm effect. Days in milk influenced milk composition, whose changes partly reflected the modifications of %CY traits. Moreover, large differences were observed between primiparous and multiparous goats: primiparous goats produced less milk of better quality (higher fat, lower somatic cell and bacterial counts) and less cheese, but with higher recovery of fat and protein in the curd, compared with multiparous goats. The repeatability was very high, for both coagulation (84.0 to 98.8%) and cheese-making traits (89.7 to 99.9%). The effect of MEL was significant for daily productions of milk and cheese, coagulation time, and recovery of protein in the curd, which were better in high-MEL farms. As regards geographic area, milk composition and percentage cheese yield were superior in the Central-East area, whereas daily milk and cheese production and MCP were better in the South-West. This result was explainable by the phenomenon of crossbreeding Sarda goats with Maltese bucks, which occurred with greater intensity in the South-West than in the Central-East area of the island. The results provided by this study could be of great interest for the goat dairy sector. Indeed, the methods described in the present study could be applicable for other farming methods, goat breeds, and geographic areas. The collection of a wide range of phenotypes at individual animal level is fundamental for the characterization of local populations and can be used to guarantee breed conservation and the persistence of traditional farming systems, and to increase farmers' profit.

Accuracy and biases in predicting the chemical and physical traits of many types of cheeses using different visible and near-infrared spectroscopic techniques and spectrum intervals.

Near-infrared spectroscopy (NIRS) has been widely used to determine various composition traits of many dairy products in the industry. In the last few years, near-infrared (NIR) instruments have become more and more accessible, and now, portable devices can be easily used in the field, allowing the direct measurement of important quality traits. However, the comparison of the predictive performances of different NIR instruments is not simple, and the literature is lacking. These instruments may use different wavelength intervals and calibration procedures, making it difficult to establish whether differences are due to the spectral interval, the chemometric approach, or the instrument's technology. Hence, the aims of this study were (1) to evaluate the prediction accuracy of chemical contents (5 traits), pH, texture (2 traits), and color (5 traits) of 37 categories of cheese; (2) to compare 3 instruments [2 benchtop, working in reflectance (R) and transmittance (T) mode (NIRS-R and NIRS-T, respectively) and 1 portable device (VisNIRS-R)], using their entire spectral ranges (1100-2498, 850-1048, and 350-1830 nm, respectively, for NIRS-R, NIRS-T and VisNIRS-R); (3) to examine different wavelength intervals of the spectrum within instrument, comparing also the common intervals among the 3 instruments; and (4) to determine the presence of bias in predicted traits for specific cheese categories. A Bayesian approach was used to develop 8 calibration models for each of 13 traits. This study confirmed that NIR spectroscopy can be used to predict the chemical composition of a large number of different cheeses, whereas pH and texture traits were poorly predicted. Color showed variable predictability, according to the trait considered, the instrument used, and, within instrument, according to the wavelength intervals. The predictive performance of the VisNIRS-R portable device was generally better than the 2 laboratory NIRS instruments, whether with the entire spectrum or selected intervals. The VisNIRS-R was found suitable for analyzing chemical composition in real time, without the need for sample uptake and processing. Our results also indicated that instrument technology is much more important than the NIR spectral range for accurate prediction equations, but the visible range is useful when predicting color traits, other than lightness. Specifically for certain categories (i.e., caprine, moldy, and fresh cheeses), dedicated calibrations seem to be needed to obtain unbiased and more accurate results.

Milk protein fractions strongly affect the patterns of coagulation, curd firming, and syneresis.

The aim of this study was to assess the role of milk protein fractions in the coagulation, curd firming, and syneresis of bovine milk. Analyses were performed on 1,271 individual milk samples from Brown Swiss cows reared in 85 herds classified into 4 types of farming systems, from the very traditional (tied cows, feed manually distributed, summer highland pasture) to the most modern (loose cows, use of total mixed rations with or without silage). Fractions αS1-casein (CN), αS2-CN, ß-CN, κ-CN, ß-lactoglobulin (LG), and α-lactalbumin (LA) and genotypes at CSN2, CSN3, and BLG were obtained by reversed-phase HPLC. The following milk coagulation properties were measured with a lactodynamograph, with the testing time extended to 60 min: rennet coagulation time (RCT, min), curd firming time (min), and curd firmness at 30 and 45 min (mm). All the curd firmness measures recorded over time (total of 240 observations/sample) were used in a 4-parameter nonlinear model to obtain parameters of coagulation, curd firming, and syneresis: RCT estimated from the equation (min), asymptotic potential curd firmness (mm), the curd firming and syneresis instant rate constants (%/min), and the maximum curd firmness value (CFmax, mm) and the time taken to reach it (min). All the aforementioned traits were analyzed with 2 linear mixed models, which tested the effects of the protein fractions expressed in different ways: in the first, quantitative model, each protein fraction was expressed as content in milk; in the second, qualitative model, each protein fraction was expressed as a percentage of total casein content. Besides proteins, additional nuisance parameters were herd (included as a random effect), daily milk production (only for the quantitative model), casein content (only for the qualitative model), dairy system, parity, days in milk, the pendulum of the lactodynamograph, and the CSN2, CSN3, and BLG genotypes. Both αS1-CN and ß-CN showed a clear and favorable effect on CFmax, where the former effect was almost double the latter. Milk coagulation ability was favorably affected by κ-CN, which reduced both the RCT and RCT estimated from the equation, increased the curd firming and syneresis instant rate constants, and allowed a higher CFmax to be reached. In contrast, αS2-CN delayed gelation time and ß-LG worsened curd firming, both resulting in a low CFmax. The results of this study suggest that modification of the relative contents of specific protein fractions can have an enormous effect on the technological behavior of bovine milk.

Effects of indirect indicators of udder health on nutrient recovery and cheese yield traits in goat milk.

In dairy goats, very little is known about the effect of the 2 most important indirect indicators of udder health [somatic cell count (SCC) and total bacterial count (TBC)] on milk composition and cheese yield, and no information is available regarding the effects of lactose levels, pH, and NaCl content on the recovery of nutrients in the curd, cheese yield traits, and daily cheese yields. Because large differences exist among dairy species, conclusions from the most studied species (i.e., bovine) cannot be drawn for all types of dairy-producing animals. The aims of this study were to quantify, using milk samples from 560 dairy goats, the contemporary effects of a pool of udder health indirect indicators (lactose level, pH, SCC, TBC, and NaCl content) on the recovery of nutrients in the curd (%REC), cheese yield (%CY), and daily cheese yields (dCY). Cheese-making traits were analyzed using a mixed model, with parity, days in milk (DIM), lactose level, pH, SCC, TBC, and NaCl content as fixed effects, and farm, breed, glass tube, and animal as random effects. Results indicated that high levels of milk lactose were associated with reduced total solids recovery in the curd and lower cheese yields, because of the lower milk fat and protein contents in samples rich in lactose. Higher pH correlated with higher recovery of nutrients in the curd and higher cheese yield traits. These results may be explained by the positive correlation between pH and milk fat, protein, and casein in goat milk. High SCC were associated with higher recovery of solids and energy in the curd but lower recovery of protein. The higher cheese yield obtained from milk with high SCC was due to both increased recovery of lactose in the curd and water retention. Bacterial count proved to be the least important factor affecting cheese-making traits, but it decreased daily cheese yields, suggesting that, even if below the legal limits, TBC should be considered in order to monitor flock management and avoid economic losses. The effect of NaCl content on milk composition was linked with lower recovery of all nutrients in the curd during cheese-making. In addition, high milk NaCl content led to reductions in fresh cheese yield and cheese solids. The indirect indicators of the present study significantly affected the cheese-making process. Such information should be considered, to adjust the milk-to-cheese economic value and the milk payment system.

Direct and indirect predictions of enteric methane daily production, yield, and intensity per unit of milk and cheese, from fatty acids and milk Fourier-transform infrared spectra.

Mitigating the dairy chain's contribution to climate change requires cheap, rapid methods of predicting enteric CH4 emissions (EME) of dairy cows in the field. Such methods may also be useful for genetically improving cows to reduce EME. Our objective was to evaluate different procedures for predicting EME traits from infrared spectra of milk samples taken at routine milk recording of cows. As a reference method, we used EME traits estimated from published equations developed from a meta-analysis of data from respiration chambers through analysis of various fatty acids in milk fat by gas chromatography (FAGC). We analyzed individual milk samples of 1,150 Brown Swiss cows from 85 farms operating different dairy systems (from very traditional to modern), and obtained the cheese yields of individual model cheeses from these samples. We also obtained Fourier-transform infrared absorbance spectra on 1,060 wavelengths (5,000 to 930 waves/cm) from the same samples. Five reference enteric CH4 traits were calculated: CH4 yield (CH4/DMI, g/kg) per unit of dry matter intake (DMI), and CH4 intensity (CH4/CM, g/kg) per unit of corrected milk (CM) from the FAGC profiles; CH4 intensity per unit of fresh cheese (CH4/CYCURD, g/kg) and cheese solids (CH4/CYSOLIDS, g/kg) from individual cheese yields (CY); and daily CH4 production (dCH4, g/d). Direct infrared (IR) calibrations were obtained by BayesB modeling; the determination coefficients of cross-validation varied from 0.36 for dCH4 to 0.57 for CH4/CM, and were similar to the coefficient of determination values of the equations based on FAGC used as the reference method (0.47 for CH4/DMI and 0.54 for CH4/CM). The models allowed us to select the most informative wavelengths for each EME trait and to infer the milk chemical features underlying the predictions. Aside from the 5 direct infrared prediction calibrations, we tested another 8 indirect prediction models. Using IR-predicted informative fatty acids (FAIR) instead of FAGC, we were able to obtain indirect predictions with about the same precision (correlation with reference values) as direct IR predictions of CH4/DMI (0.78 vs. 0.76, respectively) and CH4/CM (0.82 vs. 0.83). The indirect EME predictions based on IR-predicted CY were less precise than the direct IR predictions of both CH4/CYCURD (0.67 vs. 0.81) and CH4/CYSOLIDS (0.62 vs. 0.78). Four indirect dCH4 predictions were obtained by multiplying the measured or IR-predicted daily CM production by the direct or indirect CH4/CM. Combining recorded daily CM and predicted CH4/CM greatly increased precision over direct dCH4 predictions (0.96-0.96 vs. 0.68). The estimates obtained from the majority of direct and indirect IR-based prediction models exhibited herd and individual cow variability and effects of the main sources of variation (dairy system, parity, days in milk) similar to the reference data. Some rapid, cheap, direct and indirect IR prediction models appear to be useful for monitoring EME in the field and possibly for genetic/genomic selection, but future studies directly measuring CH4 with different breeds and dairy systems are needed to validate our findings.

Variations in milk protein fractions affect the efficiency of the cheese-making process.

The aim of this study was to assess the influence of the amounts of the αS1-, αS2-, ß-, and κ-casein (CN) and the α-lactalbumin and ß-lactoglobulin protein fractions on the efficiency of the cheese-making process independently of their genetic polymorphisms. The study was carried out on milk samples from 1,271 Brown Swiss cows from 85 herds classified into 4 categories according to management, feeding, and housing characteristics (traditional and modern systems). To assess the efficiency of the cheese-making process, we processed the milk samples according to a laboratory cheese-making procedure (1,500 mL/sample) and obtained the following measures: (1) 3 percentage cheese yields (%CYcurd, %CYsolids, %CYwater), (2) 2 daily cheese yields obtained by multiplying %CY (curd and total solids) by daily milk yields (dCYcurd, dCYsolids), (3) 4 measures of nutrient recovery in the curd (RECfat, RECprotein, RECsolids, RECenergy), and (4) 2 measures of cheese-making efficiency in terms of the ratio between the observed and theoretical %CY (Ef-%CYcurd, Ef-%CYsolids). All the aforementioned traits were analyzed by fitting 2 linear mixed models with protein fractions as fixed effects expressed as percentage in the milk (model M-%milk) and as percentage of the total casein content (model M-%cas) together with the effects of total casein content (only in model M-%cas), daily milk yield (only in model M-%milk; not for dCY traits), dairy system, herd (random effect), days in milk, parity, and vat. The efficiency of overall cheese yield (Ef-%CYcurd) was mostly positively associated with ß-CN content in the milk, whereas Ef-%CYsolids was greater with higher amounts of κ-CN and αS1-CN (M-%milk) due to the strong influence of both fractions on the recovery rate of milk components in the curd (fat and total solids, protein with αS1-CN only) when expressed as percentage of milk and of total casein; only ß-CN was more important for RECprotein. In contrast, we found ß-lactoglobulin to be highly negatively related to all the traits related to the cheese-making process and to the daily cheese yield per cow, whereas α-lactalbumin was positively associated with the latter traits. Additional research on this topic is needed, with particular focus on the genetic and genomic aspects of the role of protein fractions in the cheese-making process and on the associations between genetic polymorphisms in milk protein and milk nutrient recovery in the curd.

Phenotypic and genetic relationships between indicators of the mammary gland health status and milk composition, coagulation, and curd firming in dairy sheep.

The present study investigated the effect of somatic cell count, lactose, and pH on sheep milk composition, coagulation properties (MCP), and curd firming (CF) parameters. Individual milk samples were collected from 1,114 Sarda ewes reared in 23 farms. Milk composition, somatic cell count, single point MCP (rennet coagulation time, RCT; curd firming time, k20; and curd firmness, a30, a45, and a60), and CF model parameters were achieved. Phenotypic traits were statistically analyzed using a mixed model to estimate the effects of the different levels of milk somatic cell score (SCS), lactose, and pH, respectively. Additive genetic, herd, and residual correlations among these 3 traits, and with milk composition, MCP and CF parameters, were inferred using a Bayesian approach. From a phenotypic point of view, higher SCS levels caused a delayed gelification of milk. Lactose concentration and pH were significant for many milk quality traits, with a very intense effect on both coagulation times and curd firming. These traits (RCT, RCT estimated using the curd firming over time equation, and k20) showed an unfavorable increase of about 20% from the highest to the lowest level of lactose. Milk samples with pH values lower than 6.56 versus higher than 6.78 were characterized by an increase of RCT (from 6.00 to 14.3 min) and k20 (from 1.65 to 2.65 min) and a decrease of all the 3 curd firmness traits. From a genetic point of view, the marginal posterior distribution of heritability estimates evidenced a large and exploitable variability for all 3 phenotypes. The mean intra-farm heritability estimates were 0.173 for SCS, 0.418 for lactose content, and 0.206 for pH. Lactose (favorably), and SCS and pH (unfavorably), at phenotypic and genetic levels, were correlated mainly with RCT and RCT estimated using the curd firming over time equation and scarcely with the other curd firming traits. The SCS, lactose, and pH were significantly correlated with each other's. In conclusion, results reported in the present study suggest that SCS, pH, and lactose affect, contemporarily and independently, milk quality and MCP. These phenotypes, easily available during milk recording schemes measured by infrared spectra prediction, could be used as potential indicators traits for improving cheese-making ability of ovine milk.

Cheese yield, cheesemaking efficiency, and daily production of 6 breeds of goats.

Little is known about the complex process of cheesemaking at the individual level of dairy goats because of the difficulties of producing a high number of model cheeses. The objectives of this work were (1) to study the cheesemaking ability of goat milk; (2) to investigate the variability of cheesemaking-related traits among different farms; (3) to assess the effects of stage of lactation and parity; and (4) to compare 6 breeds of goat (Saanen and Camosciata delle Alpi for the Alpine type; Murciano-Granadina, Maltese, Sarda and Sarda Primitiva for the Mediterranean type) for their cheesemaking ability. For each goat (n = 560) we studied (1) 8 milk quality traits (fat, protein, total solids, casein, lactose, pH, somatic cell score, and bacterial count); (2) 4 milk nutrient recovery traits (fat, protein, total solids, and energy) in curd; (3) 3 actual cheese yield traits (fresh cheese, cheese solids, and cheese water); (4) 2 theoretical cheese yield values (fresh cheese and cheese solids) and the related cheesemaking efficiencies; and (5) daily milk yield and 3 daily cheese yield traits (fresh cheese, cheese solids, and water retained in the curd). With respect to individual animal factors, farm was not particularly important for recovery traits or actual and theoretical cheese yield and estimates of efficiency, whereas it highly influenced daily productions. Parity of goats influenced daily cheese production, whereas DIM slightly affected recovery as well as percent and daily cheese yield traits. Breed was the most important source of variation for almost all cheesemaking traits. Compared with those of Alpine type, the 4 Mediterranean breeds had, on average, lower daily milk and cheese productions, greater actual and theoretical cheese yield, and higher recovery of nutrients in the curd. Among Alpine type, Camosciata delle Alpi was characterized by greater nutrients recovery than Saanen. Within the 4 Mediterranean types, the 3 Italians produced much less milk per day, with much more fat and protein and greater recovery traits than the Murciano-Granadina, resulting in greater actual cheese yield. Within the Italian breeds, milk from Sarda and Sarda Primitiva was characterized by lower daily yields, higher protein and fat content, and greater recoveries of nutrients than Maltese goats. These results confirmed the potential of goat milk for cheese production and could be useful to give new possibilities and direction in breeding programs.