Luigi Gallo

Validation of a new reversed-phase high-performance liquid chromatography method for separation and quantification of bovine milk protein genetic variants

A new RP-HPLC method for the separation and quantification of the most common genetic variants of bovine milk proteins is described. A reversed-phase analytical column C8 (Zorbax 300SB-C8 RP, 3.5 microm, 300A, 150 x 4.6 I.D.) was used. All the most common casein (CN) and whey protein genetic variants, including beta-CN(I) were detected and separated simultaneously in less then 40 min, with the exception of alpha(S1)-CN(B) and CN(C) variants. Purified protein genetic variants were employed in calibration and showed different absorbances at 214 nm. The procedure was developed using 40 raw individual milk samples of cows belonging to four different breeds and certified skim milk powder BCR-063R. Method validation consisted in testing linearity, repeatability, reproducibility and accuracy. A linear relationship (R(2)>0.99) between the concentrations of proteins and peak areas was observed over the concentration range, with low detection limits. Repeatability and reproducibility were satisfactory for both retention times and peak areas. The RSD of peak areas ranged from 0.92 to 4.32% within analytical day and from 0.85 to 9.52% across analytical days. The recoveries, calculated using mixtures of samples previously quantified, ranged from 98.1 to 103.7%.

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.

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.

Genetic analysis of detailed milk protein composition and coagulation properties in Simmental cattle

The objective of this study was to estimate genetic parameters for milk protein fraction contents, milk protein composition, and milk coagulation properties (MCP). Contents of α(S1)-, α(S2)-, β-, γ-, and κ-casein (CN), β-lactoglobulin (β-LG), and α-lactalbumin (α-LA) were measured by reversed-phase HPLC in individual milk samples of 2,167 Simmental cows. Milk protein composition was measured as percentage of each CN fraction in CN (α(S1)-CN%, α(S2)-CN%, β-CN%, γ-CN%, and κ-CN%) and as percentage of β-LG in whey protein (β-LG%). Rennet clotting time (RCT) and curd firmness (a(30)) were measured by a computerized renneting meter. Heritabilities for contents of milk proteins ranged from 0.11 (α-LA) to 0.52 (κ-CN). Heritabilities for α(S1)-CN%, κ-CN%, and β-CN% were similar and ranged from 0.63 to 0.69, whereas heritability of α(S2)-CN%, γ-CN%, and β-LG% were 0.28, 0.18, and 0.34, respectively. Effects of CSN2-CSN3 haplotype and BLG genotype accounted for more than 80% of the genetic variance of α(S1)-CN%, β-CN%, and κ-CN% and 50% of the genetic variance of β-LG%. The genetic correlations among the contents of CN fractions and between CN and whey protein fractions contents were generally low. When the data were adjusted for milk protein gene effects, the magnitude of the genetic correlations among the contents of milk protein fractions markedly increased, indicating that they undergo a common regulation. The proportion of β-CN in CN correlated negatively with κ-CN% (r=-0.44). The genetic relationships between CN and whey protein composition were trivial. Low milk pH correlated with favorable MCP. Genetically, contents and proportions of α(S1)- and α(S2)-CN in CN were positively correlated with RCT. The relative proportion of β-CN in CN exhibited a genetic correlation with RCT of -0.26. Both the content and the relative proportion of κ-CN in CN did not correlate with RCT. Weak curds were genetically associated with increased proportions in CN of α(S1)- and α(S2)-CN, decreased contents of β-CN and κ-CN, and decreased proportion of κ-CN in CN. Negligible effects on the estimated correlations between a(30) and κ-CN contents or proportion in CN were observed when the model accounted for milk protein gene effects. Increasing β-CN and κ-CN contents and relative proportions in CN and decreasing the content and proportions of α(S1)-CN and α(S2)-CN and milk pH through selective breeding exert favorable effects on MCP.

Livestock systems and farming styles in Eastern Italian Alps: an on-farm survey

Abstract This research aimed to study the relationships between livestock systems, landscape maintenance and farming styles in the Belluno Province, a mountainous area of the Eastern Italian Alps. A total of 65 farms were sampled on the basis of livestock category farmed and herd size. Farms were visited to collect information on technical and productive aspects, on landscape features of land managed, which was identified by aerial photographs and digitised in a GIS environment, and on the farmers’ background, attitudes and approach to farming. Six different livestock systems were identified: intensive beef cattle (2 farms); extensive beef cattle (12 farms); large sheep/goat farms (9 farms); small sheep/goat farms (6 farms); intensive dairy cattle (14 farms) and extensive dairy cattle (22 farms). The intensive systems had larger herds, modern structures and equipment, and were strongly production oriented, whereas the extensive systems had smaller herds and productivity, with often traditional or obsolete structures and equipment, but showed a tendency to diversify production by means of on-farm cheese making and/or mixed farming of different livestock categories. The ability to maintain meadows and pastures was greater for the extensive systems, especially in steep areas, while the annual nitrogen output, estimated as kg N/ha, was lower. Data on the farmers’ background and attitudes were analysed with a non-hierarchical cluster procedure that clustered the farmers into 4 farming styles widely different in motivations to farming, innovative capability, and ability to diversify income sources and ensure farm economic viability. The farming styles were distributed across all livestock systems, indicating the lack of a linkage between the assignment of a farm to a livestock system and the way the farm is managed. This study demonstrates that in mountain areas variability of livestock systems may be high, and that they differ not only in production practices but also in the ability to maintain landscape, which is generally higher in the extensive or even marginal systems. Within a given livestock system, farms may be managed with different styles, which implies that informative knowledge to address policy decisions needs to integrate the definition of livestock systems with the assessment of farming styles.

Genetic parameters of carcass and meat quality traits of double muscled Piemontese cattle

Genetic parameters of meat quality (MQ) were estimated on Longissimus thoracis muscle of 1208 Piemontese young bulls, progeny of 109 AI sires. Carcass weight (CW), conformation (EUS) and pH (pH24h) were recorded at 24h and lightness (L*), redness (a*), yellowness (b*), pH (pH8d), drip loss (DL), cooking loss (CL) and shear force (SF) were assessed. The heritability (h(2)) of pH24h was very low (0.06), but h2 of pH8d was markedly higher (0.42). Heritability was 0.32, 0.33, and, 0.14, for L*, a* and b*, respectively, whereas was 0.24, 0.07 and 0.14, for DL, CL, and SF, respectively. The two pH measures showed opposite genetic relationships with color measures. Genetic correlations of DL and CL were positive with L* and b* and negative with a*. Genetic correlations between carcass traits and MQ suggest that animals with superior growth potential tend to exhibit reduced EUS scores and pale meat with lower tenderness and water holding capacity. Conversely, improvement of EUS score through selection would lead to light, bright, and tender meat with enhanced water holding capacity.

Survival analysis of preweaning piglet survival in a dry-cured ham-producing crossbred line

The aim of this study was to investigate piglet preweaning survival and its relationship with a total merit index (TMI) used for selection of Large White terminal boars for dry-cured ham production. Data on 13,924 crossbred piglets (1,347 litters), originated by 189 Large White boars and 328 Large White-derived crossbred sows, were analyzed under a frailty proportional hazards model, assuming different baseline hazard functions and including sire and nursed litter as random effects. Estimated hazard ratios (HR) indicated that sex, cross-fostering, year-month of birth, parity of the nurse sow, size of the nursed litter, and class of TMI were significant effects for piglet preweaning survival. Female piglets had less risk of dying than males (HR = 0.81), as well as cross-fostered piglets (HR = 0.60). Survival increased when piglets were nursed by sows of third (HR = 0.85), fourth (HR = 0.76), and fifth (HR = 0.79) parity in comparison with first and second parity sows. Piglets of small (HR = 3.90) or very large litters (HR >1.60) had less chance of surviving in comparison with litters of intermediate size. Class of TMI exhibited an unfavorable relationship with survival (HR = 1.20 for the TMI top class). The modal estimates of sire variance under different baseline hazard functions were 0.06, whereas the variance for the nursed litter was close to 0.7. The estimate of the nursed litter effect variance was greater than that of the sire, which shows the importance of the common environment generated by the nurse sow. Relationships between sire rankings obtained from different survival models were high. The heritability estimate in equivalent scale was low and reached a value of 0.03. Nevertheless, the exploitable genetic variation for this trait justifies the inclusion of piglet preweaning survival in the current breeding program for selection of Large White terminal boars for dry-cured ham production.