Uncovering genetic parameters and environmental influences on fertility, milk production, and quality in autochthonous Reggiana cattle.

Reggiana is a local cattle breed from northern Italy known for its rusticity and profitability, due to the production of branded Parmigiano Reggiano cheese. To ensure the persistence of such profitability in the long term, an adequate breeding program is required. To this aim, in the present study we estimate the genetic parameters of the main productive and reproductive traits, and we evaluate the effect of genotype by environment interaction (GxE) on these traits using 2 environmental covariates: (1) productivity and (2) temperature-humidity index (THI). Milk, fat, protein, and casein yield were considered as daily production traits, whereas protein, fat, casein percentage, casein index, and somatic cell score were considered as milk quality traits. Finally, reproductive traits such as the number of inseminations, days open, calving interval, and calving-to-first-insemination interval were evaluated. Reggiana cattle produce an average of 19 kg of milk per day with 3.7% fat and 3.4% protein content and have excellent fertility parameters. Compared with other breeds, they have slightly lower heritability for production and quality for production traits (e.g., 0.12 [0.09; 0.15] for milk yield), but similar heritability for fertility traits. Milk, protein, and fat daily yields are highly correlated but negatively correlated with the percentage of protein, fat, and casein, whereas fertility traits have an unfavorable genetic correlation with daily production traits. When considering productivity, a consistent amount of variability due to GxE was observed for all daily production traits, somatic cell count, and casein index. A modest amount of GxE was observed for fertility parameters, while the percentage of solid content showed almost no GxE effect. A similar situation occurred when considering the THI, but no GxE interaction was observed for reproduction traits. In conclusion, this study provides useful information for the implementation of accurate selection plans in this local breed, accounting for environmental plasticity measured through the consistent GxE interaction observed.

NF-YA isoforms with alternative splicing of exon-5 in Aves.

NF-YA, the regulatory subunit of the trimeric CCAAT-binding transcription factor NF-Y, is present in vertebrates in two major alternative spliced isoforms: NF-YAl and NF-YAs, differing for the presence of exon-3. NF-YAx, a third isoform without exon-3/-5, was reported only in human neuronal cells and tumors. These events affect the Trans-Activation Domain. We provide here evidence for the expression of NF-YAx and for the existence of a new isoform, NF-YAg, skipping only exon-5. These isoforms are abundant in Aves, but not in reptiles, and are the prevalent transcripts in the initial phases of embryo development in chicken. Finally, we analyzed NF-YAg and NF-YAx amino acid sequence using AlphaFold: absence of exon-5 denotes a global reduction of ß-stranded elements, while removal of the disordered exon-3 sequence has limited effects on TAD architecture. These data identify an expanded program of NF-YA isoforms within the TAD in Aves, implying a role during early development.

Economic weights for restriction of selection index as optimal strategy for combining multiple traits.

Presently, selection indices combine EBV belonging to many phenotypes accounting for production and functional traits. In this situation, ensuring positive genetic trends becomes difficult, especially when an antagonistic genetic relationship between traits exists. For this reason, guaranteeing zero genetic progress for some traits could be advantageous, as it would allow maximization of the productive characteristics while maintaining steady other antagonistic traits. This approach can also be useful for traits with an intermediate optimum, such as type traits, or for phenotypes for which it is difficult to quantify a specific economic weight. On this point, deriving the economic weight for a restricted selected index can be an optimal strategy. In this article, we provide a brief demonstration of how to derive economic weights by the restriction of some antagonistic traits. This procedure is then validated in a real-life situation using a dual-purpose cattle breed, due to the presence of more than one antagonistic trait with respect to milk yield. The R code is also provided to apply this procedure. The practical application to an actual situation demonstrates that the method produces feasible solutions in situations where more antagonistic traits are considered for selection, and prevents the detriment of some functional traits. Finally, we demonstrated how to obtain the desired restricted genetic progress when heterogeneous sources of information are present and its influence on selection response.

Recovery of n-3 polyunsaturated fatty acids and conjugated linoleic acids in ripened cheese obtained from milk of cows fed different levels of extruded flaxseed.

The aim of the study was to investigate whether the addition of extruded flaxseed (EF) in dairy cow diets had an effect on milk fat and individual fatty acids (FA) recovery in cheese after 90 d of ripening. Eighteen Holstein-Friesian cows, divided into 3 experimental groups (6 cows/group), were fed 3 isonitrogenous and isoenergetic diets with 0 (CTR), 500 (EF500), or 1,000 g/d (EF1000) of EF in 3 subsequent periods (2 wk/each), following a 3 × 3 Latin square design. Dry matter intake (DMI) and milk yield were recorded daily. Individual milk samples were collected on d 7 and 13 of each period to determine proximate and FA composition. Eighteen cheese-making sessions (2 for each group and period) were carried out, using a representative pooled milk sample obtained from the 6 cows of each group (10L). At 90 d of ripening, cheeses were analyzed for proximate and FA composition. Cheese yield was computed as the ratio between the weights of ripened cheese and processed milk. Recoveries of fat, individual FA, and grouped FA were computed as the ratio between the corresponding weights in cheese and in milk. Inclusion of EF did not affect DMI, milk yield, or milk composition. Compared with CTR, the 2 diets containing EF increased the proportion of C18:3n-3 and total n-3 FA, in both milk and cheese. Cheese yield and cheese fat percentage did not differ among diets. Likewise, milk fat recovery in cheese was comparable in the 3 treatments and averaged 0.85. The recoveries of individual FA were, for the most part, not dissimilar from fat recovery, except for short-chain saturated FA (from 0.38 for C4:0 to 0.80 for C13:0), some long-chain saturated FA (0.56 and 0.62 for C20:0 and C21:0, respectively), and for C18:3n-6 (1.65). The recovery of saturated FA was lower than that of monounsaturated FA, whereas recovery of polyunsaturated FA was intermediate. Compared with medium- and long-chain FA, short-chain FA were recovered to a smaller extent in cheese. No differences in recovery were found between n-6 and n-3 FA. In conclusion, FA have different recoveries during cheese-making, with lower values for the short-chain compared with long-chain FA, and for saturated FA compared with unsaturated FA. The addition of EF in dairy cow diets did not influence cheese yield or fat recovery in cheese, irrespective of the inclusion level. The experiment confirmed that feeding cows with EF represents a successful strategy for improving the FA profile of dairy products, through an increase of n-3 FA.

Genetic parameters for linear type traits in the Rendena dual-purpose breed.

The aim of the study was to estimate the genetic parameters for five composite traits and 20 individual type traits on 10,735 first-parity Rendena dual-purpose cows. Fixed effects included in the linear animal mixed models were herd-year-classifier, days in milk and age at first calving; the additive genetic effect of the animal was included as a random effect. Heritability estimates varied from 0.12 (feet) to 0.52 (stature). Genetic correlations between the individual body size traits were all ≥0.69; similar strong genetic correlations existed between traits describing similar morphological characteristics (e.g. mammary system, fleshiness). Many of the body size traits were negatively genetically correlated with animal fleshiness. Genetic trends showed that genetic merit for body size increased consistently over the last 10 years, while genetic merit for fleshiness declined. These results suggest that the characteristics of the dual-purpose Rendena cattle are becoming more like specialized milk-producing animals. Nonetheless, sufficient genetic variation exists to halt or reverse the deterioration in fleshiness.

Unraveling inbreeding patterns and selection signals in Alpine Grey cattle.

Inbreeding plays a crucial role in livestock breeding, influencing genetic diversity and phenotypic traits. Genomic data have helped address limitations posed by incomplete pedigrees, providing deeper insights into breed genetic diversity. This study assesses inbreeding levels via pedigree and genomic approaches and analyzes old and recent inbreeding using runs of homozygosity (ROH), and selection signals in Alpine Grey cattle. Pedigree data from 165 575 individuals, analyzed with INBUPGF90 software, computed inbreeding coefficients. Genomic-based coefficients derived from PLINK v1.9. or DetectRUNS R package analyses of 1 180 individuals' genotypes. Common single nucleotide polymorphisms within ROH pinpointed genomic regions, aggregating into "ROH islands" indicative of selection pressure. Overlaps with USCS Genome Browser unveiled gene presence. Moderate correlations (0.20-0.54) existed between pedigree and genomic coefficients, with most genomic estimators having higher (>0.8) correlation values. Inbreeding averaged 0.04 in < 8 Mb ROH segments, and 0.03 in > 16 Mb segments; > 90% of ROHs were < 8 Mb, indicating ancient inbreeding prevalence. Recent inbreeding proved less detrimental than in cosmopolitan breeds. Two major ROH islands on chromosomes 6 and 7 harbored genes linked to immune response, disease resistance (PYURF, HERC3), and fertility (EIF4EBP3, SRA1). This study underscores the need for detailed inbreeding analyses to understand genetic characteristics and historical changes in local breeds like Alpine Grey cattle. Genomic insights, especially from ROH, facilitated overcoming pedigree limitations, illuminating breed genetic diversity. Our findings reveal ancient inbreeding's enduring genetic impact and ROH islands potential for selective sweeps, elucidating traits in Alpine Grey cattle.

Maternal genetic effects throughout the life of the dam in Pirenaica beef cattle. a random regression model approach.

In beef cattle, dams play a crucial role in shaping the pre- and postnatal environment for the growth of their offspring. Acknowledging the substantial impact of maternal influence on the early development of calves, researchers utilize maternal animal models. These models take into account both maternal genetic and permanent environmental effects, operating under the assumption that these influences remain constant throughout the productive life of the cow. Nevertheless, it cannot be ruled out that these genetic and environmental effects may evolve throughout the lifespan of the cows. Therefore, this study aims to describe the changes in genetic and environmental maternal effects over the productive lifespan of cows. To accomplish this goal, we utilized random regression models, incorporating the age of the dam effect, maternal genetic effects, and environmental permanent effects using Legendre orthogonal polynomials. Additionally, the analytical model incorporated a covariate to adjust for the calf's age at recording, a two-level sex effect, a random herd-year-season effect, and an additive direct genetic effect associated linked to the calf. The dataset comprised information from dams aged between 2 and 16 years, resulting in a final database that comprised weight records of 58 332 calves from 21 673 dams. The average weight at 90 days was 135.0 ± 39.3 kg, and the mean age of the dam at calving was 7.03 ± 3.41 years. We evaluated models incorporating 2, 3, 4, 5, and 6 orthogonal polynomials alongside the standard maternal animal model. Afterward, we selected the model with five orthogonal polynomials based on the Akaike Information Criteria. The Restricted Maximum Likelihood estimates within this model indicated a direct heritability of around 0.50, and a maternal heritability ranging between 0.15 and 0.25, exhibiting a consistent increase between 4, 5 to 13 years. The genetic correlation estimates between direct and maternal genetic effects remained stable at approximately -0.55 across the lifespan of the cows. Furthermore, maternal genetic correlations between different ages of the dam decreased to around 0.7 for more distant age points. The maternal permanent correlations were notably lower, occasionally even reaching negative values, suggesting variability in environmental influence on maternal effects over the productive lifespan of the cow. Finally, the model enables the prediction of breeding values for the maternal genetic effects of the cow across its lifespan, providing opportunities for innovative selection strategies on the maternal side.

Indirect genetic effects and the genetic bases of social dominance: evidence from cattle.

Genetic studies of social behaviour have currently received new impetus from models including indirect genetic effects (IGEs) of social partners. This study aimed at investigating the contribution of conspecifics in social dominance, considered as response of dyadic interaction that is, winning (dominant individual) or losing (subordinate). A genetic correlation of -1 is expected between the attitude to win and the attitude to loose, and because a population always accounts for half winners and half losers, the heritability of the dominant status should be close to zero. Specifically, social dominance was studied in Aosta Chestnut and Aosta Black Pied (Bos taurus) breeds, alpine rustic cattle famous for traditional tournaments where pairs of cows assess dominant status in bloodless fights. The outcomes of 25,590 dyadic interactions performed by 8159 individuals in 11 years were analysed by applying a classical quantitative model and models including indirect effects. Data were analysed via Bayesian approach on a threshold trait. The assessment of variances revealed a genetic correlation of -0.976 between direct and indirect genetic components. The heritability measured on a liability scale was 0.122 for direct phenotype, but decreased to 0.014 when the total heritable variance (TBV) was considered. The trend of estimated breeding values showed that the total TBV was constant over the years, even though its direct component increased and the indirect part decreased. This result confirms the relevance of IGEs on social behaviour and the assumption that the mean individual social dominance cannot evolve within a population, due to the evolutionary constraints imposed by the 'social environment'.

Characterization of a new trabectedin-resistant myxoid liposarcoma cell line that shows collateral sensitivity to methylating agents.

Myxoid Liposarcomas (MLS), characterized by the expression of FUS-CHOP fusion gene are clinically very sensitive to the DNA binding antitumor agent, trabectedin. However, resistance eventually occurs, preventing disease eradication. To investigate the mechanisms of resistance, a trabectedin resistant cell line, 402-91/ET, was developed. The resistance to trabectedin was not related to the expression of MDR related proteins, uptake/efflux of trabectedin or GSH levels that were similar in parental and resistant cells. The 402-91/ET cells were hypersensitive to UV light because of a nucleotide excision repair defect: XPG complementation decreased sensitivity to UV rays, but only partially to trabectedin. 402-91/ET cells showed collateral sensitivity to temozolomide due to the lack of O(6) -methylguanine-DNA-methyltransferase (MGMT) activity, related to the hypermethylation of MGMT promoter. In 402-91 cells chromatin immunoprecipitation (ChIP) assays showed that FUS-CHOP was bound to the PTX3 and FN1 gene promoters, as previously described, and trabectedin caused FUS-CHOP detachment from DNA. Here we report that, in contrast, in 402-91/ET cells, FUS-CHOP was not bound to these promoters. Differences in the modulation of transcription of genes involved in different pathways including signal transduction, apoptosis and stress response between the two cell lines were found. Trabectedin activates the transcription of genes involved in the adipogenic-program such as c/EBPα and ß, in 402-91 but not in 402-91/ET cell lines. The collateral sensitivity of 402-91/ET to temozolomide provides the rationale to investigate the potential use of methylating agents in MLS patients resistant to trabectedin.

Genetic correlations of fighting ability with somatic cells and longevity in cattle.

The success in competitions may be stressful for animals and costly in terms of immune functions and longevity. Focusing on Aosta Chestnut and Aosta Black Pied cattle, selected for their fighting ability in traditional competitions, this study investigated the genetic relationships of fighting ability with udder health traits (somatic cell score and two threshold traits for somatic cells), longevity (length of productive life and number of calvings) and test-day milk, fat and protein yield. Herdbook information and phenotypic records that have been routinely collected for breeding programs in 16 years were used for the abovementioned traits. Data belonged to 9328 cows and 19 283 animals in pedigree. Single-trait animal model analyses were run using a Gibbs sampling algorithm to estimate the variance components of traits, and bivariate analyses were then performed to estimate the genetic correlations. Moderate positive genetic correlations (ra) were found for fighting ability with somatic cell score (ra=0.255), suggesting that greater fighting ability is genetically related to a detriment in udder health, in agreement with the theory. The high positive genetic correlation between fighting ability and longevity (average ra=0.669) suggests that the economic importance of fighting ability (the winning cows get an higher price at selling) had probably masked the true genetic covariances. The genetic correlation between milk yield traits and fighting ability showed large intervals, but the negative values (average ra=-0.121) agreed with previous research. This study is one of the few empirical studies on genetic correlations for the competitive success v. immune functions and longevity traits. The knowledge of the genetic correlations among productive and functional traits of interest, including fighting ability, is important in animal breeding for a sustainable genetic improvement.

Analysis of genetic correlations between beef traits in young bulls and primiparous cows belonging to the dual-purpose Rendena breed.

Selection for beef traits in Italian dual-purpose breeds is often carried out using growth and in vivo conformation recorded on young, performance tested bulls and muscularity traits scored during routinely linear type evaluation on primiparous cows. In this context, the knowledge of the genetic structure of traits obtained in different sexes and at different times is necessary for a proper selection plan. This study aimed to estimate, in the local dual-purpose Rendena breed, the genetic relationships between muscularity linear type traits from primiparous cows, the same traits scored on candidate young bulls, and the performance test traits recorded in candidate young bulls. Type traits included: front (chest and shoulder), back (loins and rump); thigh, buttocks side and rear views (two traits). Performance test traits were: average daily gain; EUROP fleshiness evaluation; and dressing percentage. Muscularity linear type traits were recorded on 11 992 first parity cows, and the muscularity type traits were scored on 957 candidate young bulls. Heritability estimates obtained for muscularity traits were moderate in young bulls (on average 0.326), about 16% higher than in primiparous cows. The average heritability for performance test traits in young bulls resulted 0.342. Moderate to strong genetic correlations were found between performance test and muscularity type traits collected in young bulls (from 0.500 between front (chest and shoulder) and average daily gain to 0.955 between thigh, buttocks side view and in vivo dressing percentage). The genetic relationships obtained between muscularity linear type traits of primiparous cows and performance traits of young bulls were variable (from a null correlation between front (chest and shoulder) and average daily gain to 0.822 between thigh, buttocks rear view and dressing percentage), with an average genetic correlation of 0.532. Generally, the traits measured during performance testing in young bulls were favourably correlated with muscularity traits evaluated on primiparous cows, indicating a common selection pathway.

Effect of extraction temperature on rheological behavior and antioxidant capacity of flaxseed gum.

Soluble flaxseed gum (SFG) extracted at different temperatures (25, 40, and 60 °C) was analyzed in relation to the yield, polysaccharides and phenolics composition, surface charge, color, and rheological properties. The yield of SFG extract increased as the extraction temperature increased. The SFG xylan was the main component regardless the extraction temperature, but a reduction of substituents on the xylose chain was observed when increasing the extraction temperature. The phenolic compounds were also affected by the extraction temperature, influencing the antioxidant capacity of the gum. For all the extraction temperatures, SFG aqueous solutions showed a shear time-independent and shear-thinning behavior. Furthermore, oscillatory measurements showed a prevailing viscous character, but the decrease of the extraction temperature resulted in an increase of both G' and G". Therefore, SFG extracted at low extraction temperatures showed higher viscous and elastic properties, while high extraction temperatures increased the antioxidant activity.

Genetic correlations among milk yield, morphology, performance test traits and somatic cells in dual-purpose Rendena breed.

Selection in native local breeds needs great carefulness due to the small population size and the risk of inbreeding. Furthermore, most breeds are dual-purpose, and milk and beef attitudes are antagonistic. For preservation purposes functional traits need to be considered. Focusing on the small local Rendena cattle, this study aimed to analyse the genetic correlations among milk, beef and udder health traits and the response to selection predicted under different scenarios. The study considered milk, fat and protein yields (MY), factor scores for udder volume (UV), conformation (UC) and muscularity obtained from type traits scored on primiparous cows, and performance test traits (PT) measured on young bulls at test station: average daily gain, in vivo SEUROP fleshiness, in vivo dressing percentage. Somatic cell score (SCS) was considered as a functional trait, with a possibility of restricting its genetic gain to zero. The study considered 281 497 MY test-day data collected on 16 974 cows, and data from linear type evaluation on 11 992 primiparous cows for factor scores. The PT data were recorded on 1428 young bulls, and SCS obtained from cell counts at milk recording. Bi-trait restricted maximum likelihood animal model analyses were performed to assess genetic parameters. Heritability varied from 0.157 (fat) to 0.442 (dressing percentage). Udder volume and MY resulted positively genetically correlated (average correlation 0.427), whereas the low-negative genetic correlation between MY and UC (-0.141) suggested a negative impact of milk gain on udder form. Beef traits of factor muscularity and PT showed medium-high favourable genetic correlations (from 0.357 to 0.984), excluding a null correlation between daily gain and muscularity. The genetic correlation MY v. muscularity was unfavourable (-0.328 on average), whereas null correlations were found in MY v. PT, apart from fat v. dressing percentage (-0.151). Somatic cell score showed low unfavourable correlations with protein (0.111) and UV (0.092), and favourable correlations with UC (-0.193). Response to selection in different scenarios indicated a good balanced gain for milk and beef when standardized economic weights of 0.66 and 0.34 are given to the two attitudes, and SCS genetic gain is restricted. Current genetic trends (MY and PT increasing, but muscularity lessening) reflect a stronger selection for milk, suggesting a slight progressive change towards a milk conformation. Aiming to preserve the dual-purpose characteristics of a breed, proper breeding policies taking into account the genetic relationships among traits and including functional traits should be applied in local dual-purpose populations.

NF-Y associates with H3-H4 tetramers and octamers by multiple mechanisms.

NF-Y is a CCAAT-binding trimer with two histonic subunits, NF-YB and NF-YC, resembling H2A-H2B. We previously showed that the short conserved domains of NF-Y efficiently bind to the major histocompatibility complex class II Ea Y box in DNA nucleosomized with purified chicken histones. Using wild-type NF-Y and recombinant histones, we find that NF-Y associates with H3-H4 early during nucleosome assembly, under conditions in which binding to naked DNA is not observed. In such assays, the NF-YB-NF-YC dimer forms complexes with H3-H4, for whose formation the CCAAT box is not required. We investigated whether they represent octamer-like structures, using DNase I, micrococcal nuclease, and exonuclease III, and found a highly positioned nucleosome on Ea, whose boundaries were mapped; addition of NF-YB-NF-YC does not lead to the formation of octameric structures, but changes in the digestion patterns are observed. NF-YA can bind to such preformed DNA complexes in a CCAAT-dependent way. In the absence of DNA, NF-YB-NF-YC subunits bind to H3-H4, but not to H2A-H2B, through the NF-YB histone fold. These results indicate that (i) the NF-Y histone fold dimer can efficiently associate DNA during nucleosome formation; (ii) it has an intrinsic affinity for H3-H4 but does not form octamers; and (iii) the interactions between NF-YA, NF-YB-NF-YC, and H3-H4 or nucleosomes are not mutually exclusive. Thus, NF-Y can intervene at different steps during nucleosome formation, and this scenario might be paradigmatic for other histone fold proteins involved in gene regulation.

The major histocompatibility complex class II Ea promoter requires TFIID binding to an initiator sequence.

The major histocompatibility complex (MHC) class II Ea promoter is dependent on the presence of conserved upstream X and Y boxes and of initiator (Inr) sequences. In vitro transcription analysis of the Inr region with linker-scanning mutants pinpoints a functionally essential element that shows homology to the terminal deoxynucleotidyltransferase (TdT) Inr; contrary to the TdT Inr and other Inrs identified so far, the key sequence, between positions +5 and +12, is located within a transcribed area. Swapping the TdT sequence into the corresponding Ea position leads to a fivefold increase in transcription rate, without altering start site selection. Inr-binding proteins LBP-1/CP2 and TIP--a TdT Inr-binding protein unrelated to YY1--recognize the Ea Inr; they interact with overlapping yet distinct sequences around the Cap site, but their binding does not coincide with Ea Inr activity. A good correlation is, rather, found with binding of immunopurified holo-TFIID to this element. TFIID interacts both with Ea TATA-like and Inr sequences, but only the latter is functionally relevant. Unlike TBP, TFIID binds in the absence of TFIIA, indicating a stabilizing role for TBP-associated factors in Ea promoter recognition. Sequence comparison with other mouse and human MHC class II promoters suggests a common mechanism of start site(s) selection for the MHC class II gene family.

A functionally essential domain of RFX5 mediates activation of major histocompatibility complex class II promoters by promoting cooperative binding between RFX and NF-Y.

Major histocompatibility complex class II (MHC-II) molecules occupy a pivotal position in the adaptive immune system, and correct regulation of their expression is therefore of critical importance for the control of the immune response. Several regulatory factors essential for the transcription of MHC-II genes have been identified by elucidation of the molecular defects responsible for MHC-II deficiency, a hereditary immunodeficiency disease characterized by regulatory defects abrogating MHC-II expression. Three of these factors, RFX5, RFXAP, and RFXANK, combine to form the RFX complex, a regulatory protein that binds to the X box DNA sequence present in all MHC-II promoters. In this study we have undertaken a dissection of the structure and function of RFX5, the largest subunit of the RFX complex. The results define two distinct domains serving two different essential functions. A highly conserved N-terminal region of RFX5 is required for its association with RFXANK and RFXAP, for assembly of the RFX complex in vivo and in vitro, and for binding of this complex to its X box target site in the MHC-II promoter. This N-terminal region is, however, not sufficient for activation of MHC-II expression. This requires an additional domain within the C-terminal region of RFX5. This C-terminal domain mediates cooperative binding between the RFX complex and NF-Y, a transcription factor binding to the Y box sequence of MHC-II promoters. This provides direct evidence that RFX5-mediated cooperative binding between RFX and NF-Y plays an essential role in the transcriptional activation of MHC-II genes.

Effects of total replacement of corn silage with sorghum silage on milk yield, composition, and quality.

BACKGROUND: In the last years, difficulties occurring in corn cultivation (i.e., groundwater shortages, mycotoxin contamination) have been forcing dairy farmers to consider alternative silages. Some experiments conducted on lactating cows have proven that the total replacement of corn silage with sorghum silage did not reduce milk yield. However, this kind of substitution involves supplementing sorghum-based diets with grains, to compensate for the lower starch content of sorghum silage compared to corn silage. Change of silage type and inclusion of starch sources in the diet would influence rumen fermentations, with possible effects on milk composition (i.e., fatty acid profile) and coagulation properties. A worsening of milk coagulation properties would have a negative economic impact in Italy, where most of the milk produced is processed into cheese. This study was designed to compare milk composition and quality, with emphasis on fatty acid profile and coagulation properties, in dairy cows fed two diets based on corn or sorghum silage. RESULTS: The sorghum diet reduced milk yield (P = 0.043) but not 4% fat corrected milk (P = 0.85). Feeding sorghum silage did not influence milk contents of protein (P = 0.07) and lactose (P = 0.65), and increased fat content (P = 0.024). No differences emerged for milk concentrations of saturated (P = 0.61) and monounsaturated fatty acids (P = 0.50), whereas polyunsaturated fatty acids were lower (P < 0.001) for the sorghum diet. Concentrations of n-6 (P < 0.001) and n-3 fatty acids (P = 0.017) were lower in milk of cows fed the sorghum diet. Milk coagulation properties did not differ between the two diets, except the "a30" (the curd firmness, expressed in mm, 30 min after rennet addition), that was lower (P = 0.042) for the sorghum diet. CONCLUSIONS: Feeding a forage sorghum silage, properly supplemented with corn meal, as total replacement of corn silage maintained milk composition and did not influence negatively milk coagulation properties, which have a great economic relevance for the Italian dairy industry. Thus, silages obtained from forage sorghums could have a potential as substitute of corn silages in dairy cow diets.

Evaluation of behaviour in stabled draught horse foals fed diets with two protein levels.

The present work is aimed at evaluating the behaviour of Italian Heavy Draught Horse (IHDH) foals reared in semi-covered stables and fed two isoenergetic total mixed rations with different dietary protein levels (13.2% and 10.6% of CP on dry matter). The study was prompted by the restrictions for nitrate emissions in farms of the European Nitrate Directive. One suggested solution is to reduce dietary protein while maintaining normal performance and welfare, but there is a lack of literature in studies of horses. The behaviours of 20 foals of 437±60 kg of BW, aged 379±37 days and stabled in four pens by sex (S) and diet (D) were video recorded and analysed to build a suitable ethogram including 18 behaviours in six categories: ingestion, resting, maintenance, movement, social activities, other. The percentage of the daily time spent in each behavioural category and single behaviours was analysed via a single traits GLM including S, D and their interaction. Daily activity was consistent with existing literature: foals spent about 33% of the day in ingestion activities and 41% in resting, whereas social interactions constituted 8% of the time and individual maintenance <2%. Concerning diet, foals fed high protein spent more time in movement (19.62±0.73% of day v. 10.45±0.73% in low-protein (LP) foals; P⩽0.001), whereas the LP group increased resting (43.42±1.12% v. 38.02±1.12%; P⩽0.001). No stereotypies were found, and daily activity followed the typical values for draught breeds for foals in both dietary groups, a result that suggests the maintenance of well-being after dietary protein reduction. This result, together with the findings of a companion study showing no changes in growth performances of foals, showed that a reduction of CP in foal diet is reconcilable with the maintenance of performance and welfare.

Cloning and characterization of the histone-fold proteins YBL1 and YCL1.

Histones are among the most conserved proteins in evolution, sharing a histone fold motif. A number of additional histonic proteins exist and are involved in the process of transcriptional regulation. We describe here the identification, cloning and characterization of two small members of the H2A-H2B sub-family (YBL1 and YCL1) related to the NF-YB and NF-YC subunits of the CCAAT-binding activator NF-Y and to the TATA-binding protein (TBP) binding repressor NC2. Unlike the latters, YBL1 and YCL1 have no intrinsic CCAAT or TATA-binding capacity. In nucleosome reconstitution assays, they can form complexes with histones in solution and on DNA and they are part of relatively large complexes, as determined by glycerol gradient experiments. Our data support the idea that YBL1 and YCL1 are divergent with respect to NF-YB and NF-YC for specific functions, but have coevolved the capacity to interact with nucleosomal structures.