Giovanni Bittante

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 (a(30), mm), and curd-firming time (k(20), 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 a(30), and samples for which k(20) 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 a(30) are highly correlated, both phenotypically and genetically. This means that the use of a(30) data does not add valuable information to that obtainable from RCT; both traits are genetically correlated mainly with milk acidity. Moreover, a(30) 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.

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 (a(30), mm), TA (SH degrees/50 mL; SH degrees = 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 (a(30), range 58 mm), 0.25 SH degrees/50 mL (TA, range 3.58 SH degrees/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 (R(2) = 0.66), whereas the predictive models developed for RCT and pH could discriminate between high and low values (R(2) = 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.

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 (a(30), 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 a(30) 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 a(30), 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 a(30) 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 a(30) 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 (a(30), 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 a(30), respectively. Reproducibility of CRM measures under different preserving conditions of milk was 93.5% for RCT and 64.6% for a(30). Repeatabilities of RCT and a(30) 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.

Effect of Holstein Friesian and Brown Swiss Breeds on Quality of Milk and Cheese

In Italy, more than 75% of milk is used for cheese making. For this reason, milk composition and coagulation traits and cheese quality represent the most important tools for the economic development of the dairy sector. In particular, cheese quality varies in relation to cheese-making technology and breed of cow. The aim of this study was to investigate the effect of 3 types of milk, originating from Holstein-Friesian (HF), Brown Swiss (BS), and mixed of both breeds, on vat milk characteristics, cheese yield, and quality in 3 different typical Italian cheese-making conditions (Casolet, Vezzena, and Grana Trentino). One hundred forty-four cows (66 HF and 78 BS) were involved, and a total of 24 vats of milk were evaluated. At maturity, 30, 21, and 16 wheels of Casolet, Vezzena, and Grana Trentino cheese were analyzed. Brown Swiss cows yielded 9% less milk per day than HF cows, but milk showed greater contents of protein, casein, titratable acidity, and better rennet coagulation time and curd firmness than HF milk. The chemical composition and cholesterol content of the 3 types of cheese were similar between breeds, whereas the cheese made with BS milk showed greater contents of monounsaturated and polyunsaturated fatty acids. Cheese made with BS milk had greater b* (yellow component) than HF. Cheese yield, recorded at different ripening times, demonstrated that BS milk yielded more cheese than HF. Mixed milk showed values, on average, intermediate to HF and BS milk characteristics, and this trend was confirmed in cheese yield at different ripening times.

Genetic parameters for conjugated linoleic acid, selected milk fatty acids, and milk fatty acid unsaturation of Italian Holstein-Friesian cows.

The objective of this study was to estimate genetic parameters for conjugated linoleic acid (CLA) and other selected milk fatty acid (FA) content and for unsaturation ratios in the Italian Holstein Friesian population. Furthermore, the relationship of milk FA with milk fat and protein content was considered. One morning milk sample was collected from 990 Italian Holstein Friesian cows randomly sampled from 54 half-sib families, located in 34 commercial herds in the North-eastern part of Italy. Each sample was analyzed for milk percentages of fat and protein, and for single FA percentages (computed as FA weight as a proportion of total fat weight). Heritabilities were moderate for unsaturated FA, ranging from 0.14 for C16:1 to 0.19 for C14:1. Less than 10% of heritability was estimated for each saturated FA content. Heritability for index of desaturation, monounsaturated FA and CLA/trans-11 18:1 ratio were 0.15, 0.14, and 0.15, respectively. Standard errors of the heritability values ranged from 0.02 to 0.06. Genetic correlations were high and negative between C16:0 and C18:0, as well as between C14:0 and C18:0. Genetic correlations of index of desaturation were high and negative with C14:0 and C16:0 (-0.70 and -0.72, respectively), and close to zero (0.03) with C18:0. The genetic correlation of C16:0 with fat percentage was positive (0.74), implying that selection for fat percentage should result in a correlated increase of C16:0, whereas trans-11 C18:1 and cis-9, trans-11 C18:2 contents decreased with increasing fat percentage (-0.69 and -0.55, respectively). Genetic correlations of fat percentage with 14:1/14 and 16:1/16 ratios were positive, whereas genetic correlations of fat percentage with 18:1/18 and CLA/trans-11 18:1 ratios were negative. These results suggest that it is possible to change the milk FA composition by genetic selection, which offers opportunities to meet consumer demands regarding health aspects of milk and dairy products.

Comparison between mechanical and near-infrared methods for assessing coagulation properties of bovine milk

The aim of the present study was to compare milk coagulation properties measured through a traditional mechanical device, the Formagraph (FRM; Foss Electric A/S, Hillerød, Denmark), and a near-infrared optical device, the Optigraph (OPT; Ysebaert SA, Frépillon, France). Individual milk samples of 913 Brown Swiss cows from 63 herds located in Trento Province (Italy) were analyzed for rennet coagulation time (RCT, min), curd-firming time (k(20), min), and 2 measures of curd firmness (a(30) and a(45),mm) using the 2 instruments and under identical conditions. The trial was performed in the same laboratory, by the same technician, and following the same procedures. Extending the analysis by either instrument to 90 min permitted RCT and k(20) values to be obtained even for late-coagulating milk samples. Milk coagulation properties measured using the OPT differed considerably from those obtained using the FRM. The average k(20) values varied greatly (8.16 vs. 5.36 min for the OPT and the FRM, respectively), as did the a(45) figures (41.49 vs. 33.66 mm for the OPT and the FRM, respectively). The proportion of noncoagulating samples for which k(20) could be estimated differed between instruments, being less for the OPT. The between-instrument correlation coefficients were either moderate (0.48 for a(30)) or low (0.24 and 0.17 for k(20) and a(45), respectively) when the same traits were compared. The correlations between k(20) and a(45), and milk yield varied among instruments, as did the correlations between k(20), a(30), and a(45) and milk composition, and the correlations between a(45) and pH. The relative influence of days in milk on k(20) and a(45) varied, as did the effect of parity on a(45) and that of the measuring unit of coagulation meter on k(20) and a(30). The RCT estimated by the OPT was the only milk coagulation property to show good agreement with the FRM-derived value, although this was not true for the data from late-coagulating samples.

Effects of Composite β- and κ-Casein Genotypes on Milk Coagulation, Quality, and Yield Traits in Italian Holstein Cows

The aim of the study was to estimate the effect of the composite CSN2 and CSN3 genotypes on milk coagulation, quality, and yield traits in Italian Holstein cows. A total of 1,042 multiparous Holstein cows reared on 34 commercial dairy herds were sampled once, concurrently with monthly herd milk recording. The data included the following traits: milk coagulation time; curd firmness; pH and titratable acidity; fat, protein, and casein contents; somatic cell score; and daily milk, fat, and protein yields. A single-trait animal model was assumed with fixed effects of herd, days in milk, parity, composite casein genotype of CSN2 and CSN3 (CSN2-CSN3), and random additive genetic effect of an animal. The composite genotype of CSN2-CSN3 showed a strong effect on both milk coagulation traits and milk and protein yields, but not on fat and protein contents and other milk quality traits. For coagulation time, the best CSN2-CSN3 genotypes were those with at least one B allele in both the CSN2 and CSN3 loci. The CSN3 locus was associated more strongly with milk coagulation traits, whereas the CSN2 locus was associated more with milk and protein yields. However, because of the tight linkage between the 2 loci, the composite genotypes, or haplotypes, are more appropriate than the single-locus genotypes if they were considered for use in selection.

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 (a(30)) 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 a(30) 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 (h(2))=0.240 and h(2)=0.210 for HF and BS, respectively] than a(30) (h(2)=0.148 and h(2)=0.168 for HF and BS, respectively). Milk coagulation properties were more heritable than a single test-day milk yield (h(2)=0.103 and h(2)=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 (h(2)=0.108). A negative genetic correlation, lower than -0.85, was estimated between RCT and a(30) 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.

Nitrogen excretion in dairy cow, beef and veal cattle, pig, and rabbit farms in Northern Italy

Abstract Reference values for N excretion of different livestock production systems are required for the application of the Nitrate Directive (91/676/EC). A survey aimed to estimate N excretion from on-farm measurements of feed consumption and performance of dairy cows (104 herds, 9,984 cows), growing cattle (40 farms, 40,157 young bulls), veal calves (34 farms, 49,206 calves), growing pigs (39 farms, 161,278 pigs) and rabbits (54 farms, 65,664 reproducing does) was conducted in Veneto from 2002 to 2003. N excretion was computed as the difference between N consumption and N retained in animal products. Dairy cow yielded 8,366 ± 1,646 kg/year of milk, consumed 6,600 ± 928 kg/year of DM, containing 2.45 ± 0.2 % DM of N, and excreted 116 ± 25 kg of N/year. No significant correlation was found between milk yield and N excretion, but the correlation between dietary N concentration and N excretion was significant (r=0.66). For growing cattle, the following mean values were achieved: daily gain 1.25 ± 0.19 kg/d; feed conversion ratio 6.9 ± 0.9 kg of DM/kg, rounds/year 1.66 ± 0.38. Nitrogen consumed, retained and excreted were, respectively, 68.7 ± 5.4, 11.4 ± 1.9 and 57.3 ± 4.9 kg/place/year. For veal calves, N consumed was 24.1 ± 1.9 kg/place/year, 12.1 ± 0.8 kg of which were retained in the body and 12.0 ± 1.5 kg were excreted. For heavy pig production, N consumed, per place and per year, averaged 19.0 ± 1.9 kg, N retained was 5.2 ± 0.5 kg and N excreted was 13.8 ± 0.4 kg. In the close-cycle rabbit farms, the doe and the relative growing rabbits (43 sold per year) consumed 11.2 ± 2.2 kg, retained 3.8 ± 0.7 kg and excreted 7.4 ± 1.5 kg N/doe/year. Nitrogen excretion estimated in this work can be considered as representative of some of the main animal production systems of the North-East of Italy. These values should not be considered as fixed, otherwise the implementation of the various strategies to reduce N excretion would not be possible. They should be considered as guidelines in the assistance both to public institutions and private enterprises in the evaluation of N excretion at farm level, favouring a more accurate quantification of the excretions, an increase of N retention efficiency and a better knowledge of the requirements of agricultural land. Moreover, a major extension of the agricultural land to be fertilised with manure should be promoted.