Genetic parameters of coagulation properties, milk yield, quality, and acidity estimated using coagulating and noncoagulating milk information in Brown Swiss and Holstein-Friesian cows

The aim of this study was to estimate heritabilities of rennet coagulation time (RCT) and curd firmness (a₃₀) and their genetic correlations with test-day milk yield, composition (fat, protein, and casein content), somatic cell score, and acidity (pH and titratable acidity) using coagulating and noncoagulating (NC) milk information. Data were from 1,025 Holstein-Friesian (HF) and 1,234 Brown Swiss (BS) cows, which were progeny of 54 HF and 58 BS artificial insemination sires, respectively. Milk coagulation properties (MCP) of each cow were measured once using a computerized renneting meter and samples not exhibiting coagulation within 31 min after rennet addition were classified as NC milk. For NC samples, RCT was unobserved. Multivariate analyses, using Bayesian methodology, were performed to estimate the genetic relationships of RCT or a₃₀ with the other traits and statistical inference was based on the marginal posterior distributions of parameters of concern. For analyses involving RCT, a right-censored Gaussian linear model was used and records of NC milk samples, being censored records, were included as unknown parameters in the model implementing a data augmentation procedure. Rennet coagulation time was more heritable [heritability (h²) = 0.240 and h² = 0.210 for HF and BS, respectively] than a₃₀ (h² = 0.148 and h² = 0.168 for HF and BS, respectively). Milk coagulation properties were more heritable than a single test-day milk yield (h² = 0.103 and h² = 0.097 for HF and BS, respectively) and less heritable than milk composition traits whose heritability ranged from 0.275 to 0.275, with the only exception of fat content of BS milk (h² = 0.108). A negative genetic correlation, lower than −0.85, was estimated between RCT and a₃₀ for both breeds. Genetic relationships of MCP with yield and composition were low or moderate and favorable. The genetic correlation of somatic cell score with RCT in BS cows was large and positive and even more positive were those of RCT with pH and titratable acidity in both breeds, ranging from 0.80 to 0.94. Including NC milk information in the data affected the estimated correlations and decreased the uncertainty associated with the estimation process. On the basis of the estimated heritabilities and genetic correlations, enhancement of MCP through selective breeding with no detrimental effects on yield and composition seems feasible in both breeds. Milk acidity may play a role as an indicator trait for indirect enhancement of MCP.     

Milk coagulation ability of five dairy cattle breeds

Samples of herd milk (506) were analyzed to assess sources of variation for milk coagulation properties (MCP) for 5 different dairy cattle breeds. Data were recorded in 55 single-breed dairy herds in the Trento province, a mountain area in northeast Italy. The 5 cattle breeds were Holstein-Friesian (8 herds), Brown Swiss (16 herds), Simmental (10 herds), Rendena (13 herds), and Alpine Gray (8 herds). Herd milk samples were analyzed for the MCP traits, milk rennet coagulation time (RCT), curd-firming time, and curd firmness (a₃₀), as well as protein and fat percentages, somatic cell count, Soxhlet-Henkel acidity, and bacterial count. An ANOVA was performed to study the effect of breed, herd within breed, DIM, month of lactation, protein and fat percentages, somatic cell score, titratable acidity, and log bacterial count within breed on MCP. Breed was the most important source of variation. In particular, the Rendena breed showed the best MCP traits at 13.5 min and 27.0 mm for RCT and a₃₀, respectively. The Holstein-Friesian breed had the worst coagulation properties at 18.0 min and 17.5 mm for RCT and a₃₀, respectively. The other 3 breeds showed intermediate coagulation properties. The RCT values were better at the beginning of lactation, whereas RCT and a₃₀ values were better in September and October (14.3 min and 25.7 mm, respectively). Among the composition traits, only the titratable acidity affected MCP traits of herd milk positively.     

Milk yield,quality, and coagulation properties of 6 breeds of goats: Environmental and individual variability

Goat milk and cheese production is continuously increasing and milk composition and coagulation properties (MCP) are useful tools to predict cheesemaking aptitude. The present study was planned to investigate the extension of lactodynamographic analysis up to 60 min in goat milk, to measure the farm and individual factors, and to investigate differences among 6 goat breeds. Daily milk yield (dMY) was recorded and milk samples collected from 1,272 goats reared in 35 farms. Goats were of 6 different breeds: Saanen and Camosciata delle Alpi for the Alpine type, and Murciano-Granadina, Maltese, Sarda, and Sarda Primitiva for the Mediterranean type. Milk composition (fat, protein, lactose, pH; somatic cell score; logarithmic bacterial count) and MCP [rennet coagulation time (RCT, min), curd-firming time (k₂₀, min), curd firmness at 30, 45, and 60 min after rennet addition (a₃₀, a₄₅, and a₆₀, mm)] were recorded, and daily fat and protein yield (dFPY g/d) was calculated as the sum of fat and protein concentration multiplied by the dMY. Data were analyzed using different statistical models to measure the effects of farm, parity_, stage of lactation and breed; lastly, the direct and the indirect effect of breed were quantified by comparing the variance of breed from models with or without the inclusion of linear regression of fat, protein, lactose, pH, bacterial, somatic cell counts, and dMY. Orthogonal contrasts were performed to compare least squares means. Almost all traits exhibited high variability, with coefficients of variation between 32 (for RCT) and 63% (for a₃₀). The proportion of variance regarding dMY, dFPY, and milk composition due to the farm was moderate, whereas for MCP it was low, except for a₆₀, at 69%. Parity affected both yield and quality traits of milk, with least squares means of dMY and dFPY showing an increase and RCT and curd firmness traits a decrease from the first to the last parity class. All milk quality traits, excluding fat, were affected by the stage of lactation; RCT and k₂₀ decreased rapidly and a₃₀ was higher from the first to the last part of lactation. Alpine breeds showed the highest dMY and dFPY but Mediterranean the best percentage of protein, fat, and lactose and a shorter k₂₀ and a greater a₃₀. Among the Mediterranean goats, Murciano-Granadina goats had the highest milk yield, fat, and protein contents, whereas Maltese, Sarda, and Sarda Primitiva were characterized by much more favorable technological properties in terms of k₂₀, a₃₀, and a₄₅. In conclusion, as both the farm and individual factors highly influenced milk composition and MCP traits, improvements of these traits should be based both on modifying management and individual goat factors. As expected, several differences were attributable to the breed effect, with the best milk production for the Alpines and milk quality and coagulation for the Mediterranean goats.     

Invited review: Genetics and modeling of milk coagulation properties

Milk coagulation properties (MCP) are conventionally measured using computerized renneting meters, mechanical or optical devices that record curd firmness over time (CF_t). The traditional MCP are rennet coagulation time (RCT, min), curd firmness (a₃₀, mm), and curd-firming time (k₂₀, min). The milk of different ruminant species varies in terms of CF_t pattern. Milk from Holstein-Friesian and some Scandinavian cattle breeds yields higher proportions of noncoagulating samples, samples with longer RCT and lower a₃₀, and samples for which k₂₀ is not estimable, than does milk from Brown Swiss, Simmental, and other local Alpine breeds. The amount, proportion, and genetic variants (especially κ-casein) of milk protein fractions strongly influence MCP and explain variable proportions of the observed differences among breeds and among individuals of the same breed. In addition, other major genes have been shown to affect MCP. Individual repeatability of MCP is high, whereas any herd effect is low; thus, the improvement of MCP should be based principally on selection. Exploitable additive genetic variation in MCP exists and has been assessed using different breeds in various countries. Several models have been formulated that either handle noncoagulating samples or not. The heritability of MCP is similar to that of other milk quality traits and is higher than the heritability of milk yield. Rennet coagulation time and a₃₀ are highly correlated, both phenotypically and genetically. This means that the use of a₃₀ data does not add valuable information to that obtainable from RCT; both traits are genetically correlated mainly with milk acidity. Moreover, a₃₀ is correlated with casein content. The major limitations of traditional MCP can be overcome by prolonging the observation period and by using a novel CF_t modeling, which uses all available information provided by computerized renneting meters and allows the estimation of RCT, the potential asymptotic curd firmness, the curd-firming rate, and the syneresis rate. Direct measurements of RCT obtained from both mechanical and optical devices show similar heritabilities and exhibit high phenotypic and genetic correlations. Moreover, mid-infrared reflectance spectroscopy can predict MCP. The heritabilities of predicted MCP are higher than those of measured MCP, and the 2 sets of values are strongly correlated. Therefore, mid-infrared reflectance spectroscopy is a reliable and cheap method whereby MCP can be improved at the population level; this is because such spectra are already routinely acquired from the milk of cows enrolled in milk recording schemes.     

Short communication: Factors affecting coagulation properties of Mediterranean buffalo milk

The aim of this study was to investigate sources of variation of milk coagulation properties (MCP) of buffalo cows. Individual milk samples were collected from 200 animals in 5 herds located in northern Italy from January to March 2010. Rennet coagulation time (RCT, min) and curd firmness after 30 min from rennet addition (a₃₀, mm) were measured using the Formagraph instrument (Foss Electric, Hillerød, Denmark). In addition to MCP, information on milk yield, fat, protein, and casein contents, pH, and somatic cell count (SCC) was available. Sources of variation of RCT and a₃₀ were investigated using a linear model that included fixed effects of herd, days in milk (DIM), parity, fat content, casein content (only for a₃₀), and pH. The coefficient of determination was 51% for RCT and 48% for a₃₀. The most important sources of variation of MCP were the herd and pH effects, followed by DIM and fat content for RCT, and casein content for a₃₀. The relevance of acidity in explaining the variation of both RCT and a₃₀, and of casein content in explaining that of a₃₀, confirmed previous studies on dairy cows. Although future research is needed to investigate the effect of these sources of variation on cheese yield, findings from the present study suggest that casein content and acidity may be used as indicator traits to improve technological properties of buffalo milk.     

Short communication: Influence of composite casein genotypes on additive genetic variation of milk production traits and coagulation properties in Holstein-Friesian cows

The aim of the study was to quantify the effects of composite β- and κ-casein (CN) genotypes on genetic variation of milk coagulation properties (MCP); milk yield; fat, protein, and CN contents; somatic cell score; pH; and titratable acidity (TA) in 1,042 Italian Holstein-Friesian cows. Milk coagulation properties were defined as rennet coagulation time (RCT) and curd firmness (a₃₀). Variance components were estimated using 2 animal models: model 1 included herd, days in milk, and parity as fixed effects and animal and residual as random effects, and model 2 was model 1 with the addition of composite β- and κ-CN genotype as a fixed effect. Genetic correlations between RCT and a₃₀ and between these traits and milk production traits were obtained with bivariate analyses, based on the same models. The inclusion of casein genotypes led to a decrease of 47, 68, 18, and 23% in the genetic variance for RCT, a₃₀, pH, and TA, respectively, and less than 6% for other traits. Heritability of RCT and a₃₀ decreased from 0.248 to 0.143 and from 0.123 to 0.043, respectively. A moderate reduction was found for pH and TA, whereas negligible changes were detected for other milk traits. Estimates of genetic correlations were comparable between the 2 models. Results show that composite β- and κ-CN genotypes are important for RCT and a₃₀ but cannot replace the recording of MCP themselves.     

Genetic parameters of milk coagulation properties and their relationships with milk yield and quality traits in Italian Holstein cows

Milk coagulation properties (MCP) are an important aspect in assessing cheese-making ability. Several studies showed that favorable conditions of milk reactivity with rennet, curd formation rate, and curd strength, as well as curd syneresis, have a positive effect on the entire cheese-making process and subsequently on the ripening of cheese. Moreover, MCP were found to be heritable, but little scientific literature is available about their genetic aspects. The aims of this study were to estimate heritability of MCP and genetic correlations among MCP and milk production and quality traits. A total of 1,071 Italian Holstein cows (progeny of 54 sires) reared in 34 herds in Northern Italy were sampled from January to July 2004. Individual milk samples were collected during the morning milking and analyzed for coagulation time (RCT), curd firmness (a₃₀), pH, titratable acidity, fat, protein, and casein contents, and somatic cell count. About 10% of individual milk samples did not coagulate in 31 min, so they were removed from the analyses. Estimates of heritability for RCT and a₃₀ were 0.25 ± 0.04 and 0.15 ± 0.03, respectively. Estimates of genetic correlations between MCP traits and milk production traits were negligible except for a₃₀ with protein and casein contents (0.44 ± 0.10 and 0.53 ± 0.09, respectively). Estimates of genetic correlations between MCP traits and somatic cell score were strong and favorable, as well as those between MCP and pH and titratable acidity. Selecting for high casein content, milk acidity, and low somatic cell count might be an indirect way to improve MCP without reducing milk yield and quality traits.     

Mid-infrared spectroscopy predictions as indicator traits in breeding programs for enhanced coagulation properties of milk

The aims of this study were to investigate variation of milk coagulation property (MCP) measures and their predictions obtained by mid-infrared spectroscopy (MIR), to investigate the genetic relationship between measures of MCP and MIR predictions, and to estimate the expected response from a breeding program focusing on the enhancement of MCP using MIR predictions as indicator traits. Individual milk samples were collected from 1,200 Brown Swiss cows (progeny of 50 artificial insemination sires) reared in 30 herds located in northern Italy. Rennet coagulation time (RCT, min) and curd firmness (a₃₀, mm) were measured using a computerized renneting meter. The MIR data were recorded over the spectral range of 4,000 to 900 cm⁻¹. Prediction models for RCT and a₃₀ based on MIR spectra were developed using partial least squares regression. A cross-validation procedure was carried out. The procedure involved the partition of available data into 2 subsets: a calibration subset and a test subset. The calibration subset was used to develop a calibration equation able to predict individual MCP phenotypes using MIR spectra. The test subset was used to validate the calibration equation and to estimate heritabilities and genetic correlations for measured MCP and their predictions obtained from MIR spectra and the calibration equation. Point estimates of heritability ranged from 0.30 to 0.34 and from 0.22 to 0.24 for RCT and a₃₀, respectively. Heritability estimates for MCP predictions were larger than those obtained for measured MCP. Estimated genetic correlations between measures and predictions of RCT were very high and ranged from 0.91 to 0.96. Estimates of the genetic correlation between measures and predictions of a₃₀ were large and ranged from 0.71 to 0.87. Predictions of MCP provided by MIR techniques can be proposed as indicator traits for the genetic enhancement of MCP. The expected response of RCT and a₃₀ ensured by the selection using MIR predictions as indicator traits was equal to or slightly less than the response achievable through a single measurement of these traits. Breeding strategies for the enhancement of MCP based on MIR predictions as indicator traits could be easily and immediately implemented for dairy cattle populations where routine acquisition of spectra from individual milk samples is already performed.     

Reproducibility and repeatability of measures of milk coagulation properties and predictive ability of mid-infrared reflectance spectroscopy

The objectives of the study were to estimate the reproducibility and repeatability of milk coagulation properties (MCP) measured by a computerized renneting meter (CRM) and to evaluate the predictive ability of mid-infrared spectroscopy (MIRS) as an innovative technology for the assessment of rennet coagulation time (RCT, min) and curd firmness (a₃₀, mm). Four samples without addition of preservative (NP) and 4 samples with Bronopol addition (PS) were collected from each of 83 Holstein-Friesian cows. Six hours after collection, 2 replicated measures of MCP were obtained with CRM using 1 NP and 1 PS sample from each cow. Mid-infrared spectra of the remaining NP and PS samples from each animal were recorded after 6 h, 4 d, and 8 d after sampling. Two groups of calibration equations were developed using MIRS spectra and CRM measures of MCP as reference data obtained from analysis of NP and PS, respectively. Reproducibility and repeatability of CRM measures were obtained from REML estimation of variance components on the basis of a linear model including the fixed effects of herd and days in milk class and the random effects of cows, sample treatment (addition or no addition of preservative), and the interaction between cow and sample treatment. Coefficient of reproducibility is an indicator of the agreement between 2 measurements of MCP for the same milk sample preserved with or without addition of Bronopol. Coefficient of repeatability is an indicator of the agreement between repeated measures of MCP. Pearson correlations between MCP measures for NP and PS were 0.97 and 0.83 for RCT and a₃₀, respectively. Reproducibility of CRM measures under different preserving conditions of milk was 93.5% for RCT and 64.6% for a₃₀. Repeatabilities of RCT and a₃₀ measures were 95.7 and 77.3%, respectively. Based on the estimated cross-validation standard errors and coefficients of determination and ratios of standard errors of cross-validation to standard deviation of reference data, the predictive ability of MIRS calibration equations was moderate for RCT and unsatisfactory for a_30. Predictive ability of equations based on spectra and MCP measures of PS was greater than that of equations based on data of NP. The study did not provide conclusive evidence on the effectiveness of MIRS as a predictive tool for MCP and it requires an enlargement of the variability of milk sampling circumstances. Because the relevance of MIRS predictions in relation to breeding programs for MCP based on indicator traits relies on the genetic variation of MIRS predictions and on phenotypic and genetic correlations between MIRS predictions and MCP measures, additional specific investigations on these topics are needed.     

Short communication: Genetic correlation and heritability of milk coagulation traits within and across lactations in Holstein cows using multiple-lactation random regression animal models

Genetic parameters of milk rennet coagulation time (RCT) and curd firmness (a₃₀) among the first 3 lactations in Holstein cows were estimated. The data set included 39,960 test-day records from 5,216 Estonian Holstein cows (the progeny of 306 sires), which were recorded from April 2005 to May 2010 in 98 herds across the country. A multiple-lactation random regression animal model was used. Individual milk samples from each cow were collected during routine milk recording. These samples were analyzed for milk composition and coagulation traits with intervals of 2 to 3 mo in each lactation (7 to 305 DIM) and from first to third lactation. Mean heritabilities were 0.36, 0.32, and 0.28 for log-transformed RCT [ln(RCT)] and 0.47, 0.40, and 0.62 for a₃₀ for parities 1, 2, and 3, respectively. Mean repeatabilities for ln(RCT) were 0.53, 0.55, and 0.56, but 0.59, 0.61, and 0.68 for a₃₀ for parities 1, 2 and 3, respectively. Mean genetic correlations between ln(RCT) and a₃₀ were −0.19, −0.14, and 0.02 for parities 1, 2, and 3, respectively. Mean genetic correlations were 0.91, 0.79, and 0.99 for ln(RCT), and 0.95, 0.94, and 0.94 for a₃₀ between parities 1 and 2, 1 and 3, and 2 and 3, respectively. Due to these high genetic correlations, we concluded that for a proper genetic evaluation of milk coagulation properties it is sufficient to record RCT and a₃₀ only in the first lactation.     

Repeatability estimates for milk coagulation traits and non-coagulation of milk in Finnish Ayrshire cows

Effects of systematic environmental factors and milk production and quality traits on milk coagulation properties (MCP), and on repeatability of those traits were estimated from 979 milk samples collected once a month over a period of 2 years from 83 Finnish Ayrshire cows. Estimation was based on a multitrait animal model and REML methodology. In addition, persistence of non-coagulation of milk in individual cows, and factors associated with it were established from a sub sample of 24 cows producing non-coagulating (NC) milk at least once. MCP were at their best during the first lactation, at the beginning and at the end of lactation, and during grazing seasons. Variation in MCP with systematic environmental factors was partly due to variation in composition and quality of milk, especially in pH and ln (somatic cell count, SCC). Coefficients of repeatability for milk coagulation time and curd firmness were 0.65 and 0.68. These estimates were of the same magnitude as those for protein content, but were higher than those for daily milk yield, fat content, pH, and SCC. Based on the repeatability estimates for the milk coagulation traits and effects of the environmental factors, cows should be sampled at least three times during a lactation to estimate reliably breeding values for the milk coagulation traits. A total of 10% of the milk samples did not coagulate in 30 min after addition of rennet. Cows that produced NC milk at least once (30% of the cows) could be classified into those that produced NC milk only a few times during a lactation and those that produced NC milk at almost every sampling. Based on logistic regression analyses, peak and mid-lactation, high milk yield, low protein and fat content and high pH increased the risk of non-coagulation of milk.     

Effect of microparticulated whey proteins on milk coagulation properties

The enhancement of milk coagulation properties (MCP) and the reuse of whey produced by the dairy industry are of great interest to improve the efficiency of the cheese-making process. Native whey proteins (WP) can be aggregated and denatured to obtain colloidal microparticulated WP (MWP). The objective of this study was to assess the effect of MWP on MCP; namely, rennet coagulation time (RCT), curd-firming time, and curd firmness 30 min after rennet addition. Six concentrations of MWP (vol/vol; 1.5, 3.0, 4.5, 6.0, 7.5, and 9.0%) were added to 3 bulk milk samples (collected and analyzed during 3 d), and a sample without MWP was used as control. Within each day of analysis, 6 replicates of MCP for each treatment were obtained, changing the position of the treatment in the rack. For control samples, 2 replicates per day were performed. In addition to MCP, WP fractions were measured on each treatment during the 3 d of analysis. Milk coagulation properties were measured on 144 samples by using a Formagraph (Foss Electric, Hillerød, Denmark). Increasing the amount of MWP added to milk led to a longer RCT. In particular, significant differences were found between RCT of the control samples (13.5 min) and RCT of samples with 3.0% (14.6 min) or more MWP. A similar trend was observed for curd-firming time, which was shortest in the control samples and longest in samples with 9.0% MWP (21.4 min). No significant differences were detected for curd firmness at 30 min across concentrations of MWP. Adjustments in cheese processing should be made when recycling MWP, in particular during the coagulation process, by prolonging the time of rennet activity before cutting the curd.     

Phenotypic factors affecting coagulation properties of milk from Sarda ewes

In this study, milk-coagulation properties (MCP) were characterized in the Sarda sheep breed. Milk composition and MCP [rennet-coagulation time (RCT), curd-firming time [time to reach a curd firmness of 20 mm (k₂₀)], and curd firmness (a₃₀), (a₄₅), and (a₆₀)] were obtained extending the lactodynamographic analysis from 30 to 60 min from a population of 1,121 ewes from 23 different farms. Managerial characteristics of farms and parity, individual daily milk yields and stage of lactation of ewes were recorded. Data were analyzed using a mixed-model procedure with fixed effects of days in milk, parity, daily milk yield, and flock size and the random effect of the flock/test day nested within flock size. Sampled farms were classified as small (<300 ewes) and medium (300 to 600 ewes), and these were kept by family operations, or as large (>600 ewes), often operated through hired workers. Daily milk yield was, on average, 1.58 ± 0.79 L/d and variability for this trait was very high. The average content of fat, protein, and casein was respectively 6.41, 5.39, and 4.20%. The class of flock size had a significant effect only on curd firmness, whereas days in milk affected RCT and k₂₀. The flock test day, parity, and daily milk yield were important sources of variation for all MCP. The mean value of RCT (8.6 min) and the low occurrence of noncoagulating samples (0.44%) confirmed the excellent coagulation ability of sheep milk compared with cattle milk. A more rapid coagulation was observed in mid-lactating, primiparous, and high-yielding ewes. The k₂₀ was usually reached in less than 2 min after gelation, with the most favorable values at mid lactation. The mean value of curd firmness 30 min after rennet addition (a₃₀) was, on average, 50 mm and decreased to 46 and 42 mm respectively after 45 (a₄₅) and 60 min (a₆₀). The decreasing value of curd-firmness traits was likely to be caused by curd syneresis and whey expulsion. The correlation between RCT and a₃₀ was much lower than in dairy cows and about null for a₄₅ and a₆₀. This means that curd firmness in dairy ewes is almost independent of gelation time and this can provide specific information for this species. In conclusion, this study showed that milk from Sarda sheep is characterized by an earlier gelation, a faster increase in curd firmness with time, and greater curd firmness after 30 min compared with dairy cows. Furthermore, correlations between MCP in sheep are much lower than in bovines and some of the assumptions and interpretations related to cows cannot be applied to sheep.     

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 60 min (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 (RCT_eq), asymptotic potential curd firmness (CF_P), curd firming instant rate constant (k_CF), and syneresis instant rate constant (k_SR) and 2 derived traits: maximum curd firmness achieved within 45 min (CF_max) and time at achievement of CF_max (t_max) 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 t_max to 82.5% for RCT_eq. The repeatability of MCP was high (>80%) for all traits except those associated with the last part of the lactodynamographic curve (i.e., a₆₀, k_SR, k_CF, and t_max: 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 (RCT_eq: 18.6 vs. 16.3 min) but greater potential curd firmness (CF_P: 76.8 vs. 71.9 mm) 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.     

Prediction of coagulation properties, titratable acidity, and pH of bovine milk using mid-infrared spectroscopy

This study investigated the potential application of mid-infrared spectroscopy (MIR 4,000-900 cm⁻¹) for the determination of milk coagulation properties (MCP), titratable acidity (TA), and pH in Brown Swiss milk samples (n = 1,064). Because MCP directly influence the efficiency of the cheese-making process, there is strong industrial interest in developing a rapid method for their assessment. Currently, the determination of MCP involves time-consuming laboratory-based measurements, and it is not feasible to carry out these measurements on the large numbers of milk samples associated with milk recording programs. Mid-infrared spectroscopy is an objective and nondestructive technique providing rapid real-time analysis of food compositional and quality parameters. Analysis of milk rennet coagulation time (RCT, min), curd firmness (a₃₀, mm), TA (SH°/50 mL; SH° = Soxhlet-Henkel degree), and pH was carried out, and MIR data were recorded over the spectral range of 4,000 to 900 cm⁻¹. Models were developed by partial least squares regression using untreated and pretreated spectra. The MCP, TA, and pH prediction models were improved by using the combined spectral ranges of 1,600 to 900 cm⁻¹, 3,040 to 1,700 cm⁻¹, and 4,000 to 3,470 cm⁻¹. The root mean square errors of cross-validation for the developed models were 2.36 min (RCT, range 24.9 min), 6.86 mm (a₃₀, range 58 mm), 0.25 SH°/50 mL (TA, range 3.58 SH°/50 mL), and 0.07 (pH, range 1.15). The most successfully predicted attributes were TA, RCT, and pH. The model for the prediction of TA provided approximate prediction (R² = 0.66), whereas the predictive models developed for RCT and pH could discriminate between high and low values (R² = 0.59 to 0.62). It was concluded that, although the models require further development to improve their accuracy before their application in industry, MIR spectroscopy has potential application for the assessment of RCT, TA, and pH during routine milk analysis in the dairy industry. The implementation of such models could be a means of improving MCP through phenotypic-based selection programs and to amend milk payment systems to incorporate MCP into their payment criteria.     

Modeling rennet coagulation time and curd firmness of milk

Milk coagulation properties (MCP) are traditionally expressed using rennet coagulation time (RCT), time to curd firmness (CF) of 20 mm (k₂₀), and CF 30 min after enzyme addition (a₃₀) values, all of which are single-point measures taken from the output of computerized renneting meters, such as the Formagraph. Thus, traditional MCP use only some of the available information. Moreover, because of the worldwide spreading of breeds such as the Holstein-Friesian, characterized by late-coagulating milk, it happens often that some samples do not coagulate at all, that a₃₀ is strongly and negatively related to RCT, and that k₂₀ is not measurable. The aim of the present work was to model CF as a function of time (CF_t, mm) over a 30-min interval. The model tested was CFt=CFP×(1−e−kCF×(t−RCT)), where CF_P (mm) is the potential asymptotical CF at an infinite time, k_CF (min⁻¹) is the curd firming rate constant, and RCT is measured in minutes. The CF_t model was initially applied to data of milk of each of 105 Brown Swiss cows from 7 herds, each sampled once (trial 1). Four samples did not coagulate within 30 min. Eighty-seven of the 101 individual equations obtained fit the CF data of milk samples very well, even though the samples differed in composition, and were produced by cows of different ages and days in milk, reared on different farms (coefficient of determination >0.99; average residual standard deviation = 0.21 mm). Samples with a very late RCT (slowly coagulating samples) yielded so few observational data points that curve parameters could not be precisely estimated. The repeatability of CF_t equation parameters was estimated using data obtained from 5 replicates of each of 2 samples of bulk milk from 5 Holstein-Friesian cows analyzed every day for 5 consecutive days (trial 2). Repeatability of RCT was better than that of the other 2 parameters. Moreover, traditional MCP values (RCT, a₃₀, and k₂₀) can be obtained from the individual CF_t equations, using all available information. The MCP estimated from equations were very similar to the single-point measures yielded by the computerized renneting meter (coefficient of determination >0.97), but repeatability was slightly better. The model allowed the estimation of k₂₀ for samples with a very late coagulation or with very slow curd firming. Finally, the 3 novel parameters used to assess different milk samples were less interdependent than are the traditional measures, and their practical and scientific utility requires further study.     

Selection for milk coagulation properties predicted by Fourier transform infrared spectroscopy in the Italian Holstein-Friesian breed

Milk coagulation is based on a series of physicochemical changes at the casein micelle level, resulting in formation of a gel. Milk coagulation properties (MCP) are relevant for cheese quality and yield, important factors for the dairy industry. They are also evaluated in herd bulk milk to reward or penalize producers of Protected Designation of Origin cheeses. The economic importance of improving MCP justifies the need to account for this trait in the selection process. A pilot study was carried out to determine the feasibility of including MCP in the selection schemes of the Italian Holstein. The MCP were predicted in 1,055 individual milk samples collected in 16 herds (66 ± 24 cows per herd) located in Brescia province (northeastern Italy) by means of Fourier transform infrared (FTIR) spectroscopy. The coefficient of determination of prediction models indicated moderate predictions for milk rennet coagulation time (RCT = 0.65) and curd firmness (a₃₀ = 0.68), and poor predictions for curd-firming time (k₂₀ = 0.49), whereas the range error ratio (8.9, 6.9, and 9.5 for RCT, k₂₀, and a₃₀, respectively) indicated good practical utility of the predictive models for all parameters. Milk proteins were genotyped and casein haplotypes (α_S1-, β-, α_S2-, and κ-casein) were reconstructed. Data from 51 half-sib families (19.9 ± 16.4 daughters per sire) were analyzed by an animal model to estimate (1) the genetic parameters of predicted RCT, k₂₀, and a₃₀; (2) the breeding values for these predicted clotting variables; and (3) the effect of milk protein genotypes and casein haplotypes on predicted MCP (pMCP). This is the first study to estimate both genetic parameters and breeding values of pMCP, together with the effects of milk protein genotypes and casein haplotypes, that also considered k₂₀, probably the most important parameter for the dairy industry (because it indicates the time for the beginning of curd-cutting). Heritability of predicted RCT (0.26) and k₂₀ (0.31) were close to the average heritability described in literature, whereas the heritability of a₃₀ was higher (0.52 vs. 0.27). The effects of milk proteins were statistically significant and similar to those obtained on measured MCP. In particular, haplotypes including uncommon variants showed positive (B-I-A-B) or negative (B-A¹-A-E) effects. Based on these findings, FTIR spectroscopy-pMCP is proposed as a potential selection criterion for the Italian Holstein.     

Pathway-based genome-wide association analysis of milk coagulation properties,curd firmness,cheese yield,and curd nutrient recovery in dairy cattle

It is becoming common to complement genome-wide association studies (GWAS) with gene-set enrichment analysis to deepen the understanding of the biological pathways affecting quantitative traits. Our objective was to conduct a gene ontology and pathway-based analysis to identify possible biological mechanisms involved in the regulation of bovine milk technological traits: coagulation properties, curd firmness modeling, individual cheese yield (CY), and milk nutrient recovery into the curd (REC) or whey loss traits. Results from 2 previous GWAS studies using 1,011 cows genotyped for 50k single nucleotide polymorphisms were used. Overall, the phenotypes analyzed consisted of 3 traditional milk coagulation property measures [RCT: rennet coagulation time defined as the time (min) from addition of enzyme to the beginning of coagulation; k₂₀: the interval (min) from RCT to the time at which a curd firmness of 20 mm is attained; a₃₀: a measure of the extent of curd firmness (mm) 30 min after coagulant addition], 6 curd firmness modeling traits [RCT_eq: RCT estimated through the CF equation (min); CF_P: potential asymptotic curd firmness (mm); k_CF: curd-firming rate constant (% × min^?1); k_SR: syneresis rate constant (% × min^?1); CF_max: maximum curd firmness (mm); and t_max: time to CF_max (min)], 3 individual CY-related traits expressing the weight of fresh curd (%CY_CURD), curd solids (%CY_SOLIDS), and curd moisture (%CY_WATER) as a percentage of weight of milk processed and 4 milk nutrient and energy recoveries in the curd (REC_FAT, REC_PROTEIN, REC_SOLIDS, and REC_ENERGY calculated as the % ratio between the nutrient in curd and the corresponding nutrient in processed milk), milk pH, and protein percentage. Each trait was analyzed separately. In total, 13,269 annotated genes were used in the analysis. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway databases were queried for enrichment analyses. Overall, 21 Gene Ontology and 17 Kyoto Encyclopedia of Genes and Genomes categories were significantly associated (false discovery rate at 5%) with 7 traits (RCT, RCT_eq, k_CF, %CY_SOLIDS, REC_FAT, REC_SOLIDS, and REC_ENERGY), with some being in common between traits. The significantly enriched categories included calcium signaling pathway, salivary secretion, metabolic pathways, carbohydrate digestion and absorption, the tight junction and the phosphatidylinositol pathways, as well as pathways related to the bovine mammary gland health status, and contained a total of 150 genes spanning all chromosomes but 9, 20, and 27. This study provided new insights into the regulation of bovine milk coagulation and cheese ability that were not captured by the GWAS.     

Phenotypic and genetic characterization of the occurrence of noncoagulating milk in dairy sheep

Milk coagulation ability is of central importance for the sheep dairy industry because almost all sheep milk is destined for cheese processing. The occurrence of milk with impaired coagulation properties is an obstacle to cheese processing and, in turn, to the profitability of the dairy companies. In this work, we investigated the causes of noncoagulation of sheep milk; specifically, we studied the effect of milk physicochemical properties on milk coagulation status [coagulating and noncoagulating (NC) milk samples, which do or do not coagulate within 30 min, respectively], and whether mid-infrared spectroscopy (MIR) could be used to assess variability in coagulation status. We also investigated the genetic background of milk coagulation ability. Individual milk samples were collected from 996 Sarda ewes farmed in 47 flocks located in Sardinia (Italy). Considered traits were daily milk yield, milk composition traits, and milk coagulation properties (rennet coagulation time, curd firming time, and curd firmness), and MIR spectra were acquired. About 9% of samples did not coagulate within 30 min. A logistic regression approach was used to test the effect of milk-related traits on milk coagulation status. A principal component (PC) analysis was carried out on the milk MIR spectra, and PC scores were then used as covariates in a logistic regression model to assess their relationship with milk coagulation status. Results of the present work demonstrated that the probability of having NC samples increases as milk contents of proteins and chlorides and somatic cell score increase. The analysis of PC extracted from milk spectra that influenced coagulation status highlighted key regions associated with lactose and protein concentrations, and others not associated with routinely collected milk composition traits. These results suggest that the occurrence of NC is mostly related to damage of the epithelium secretory mammary cells, which occurs with the advancement of a lactation or due to unhealthy mammary gland status. Genetic analysis of milk coagulation status and of the extracted PC confirmed the genetic background of the milk coagulability of sheep milk.     

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.