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Abstract Background Rendena is a dual-purpose cattle breed, which is primarily found in the Italian Alps and the eastern areas of the Po valley, and recognized for its longevity, fertility, disease resistance and adaptability to steep Alpine pastures. It is categorized as 'vulnerable to extinction' with only 6057 registered animals in 2022, yet no comprehensive analyses of its molecular diversity have been performed to date. The aim of this study was to analyse the origin, genetic diversity, and genomic signatures of selection in Rendena cattle using data from samples collected in 2000 and 2018, and shed light on the breed's evolution and conservation needs. Results Genetic analysis revealed that the Rendena breed shares genetic components with various Alpine and Po valley breeds, with a marked genetic proximity to the Original Braunvieh breed, reflecting historical restocking efforts across the region. The breed shows signatures of selection related to both milk and meat production, environmental adaptation and immune response, the latter being possibly the result of multiple rinderpest epidemics that swept across the Alps in the eighteenth century. An analysis of the Rendena cattle population spanning 18 years showed an increase in the mean level of inbreeding over time, which is confirmed by the mean number of runs of homozygosity per individual, which was larger in the 2018 sample. Conclusions The Rendena breed, while sharing a common origin with Brown Swiss, has developed distinct traits that enable it to thrive in the Alpine environment and make it highly valued by local farmers. Preserving these adaptive features is essential, not only for maintaining genetic diversity and enhancing the ability of this traditional animal husbandry to adapt to changing environments, but also for guaranteeing the resilience and sustainability of both this livestock system and the livelihoods within the Rendena valley.

Abstract Background The gut microbiome plays a crucial role in understanding complex biological mechanisms, including host resilience to stressors. Investigating the microbiota-resilience link in animals and plants holds relevance in addressing challenges like adaptation of agricultural species to a warming environment. This study aims to characterize the microbiota-resilience connection in swine. As resilience is not directly observable, we estimated it using four distinct indicators based on daily feed consumption variability, assuming animals with greater intake variation may face challenges in maintaining stable physiological status. These indicators were analyzed both as linear and categorical variables. In our first set of analyses, we explored the microbiota-resilience link using PERMANOVA, α-diversity analysis, and discriminant analysis. Additionally, we quantified the ratio of estimated microbiota variance to total phenotypic variance (microbiability). Finally, we conducted a Partial Least Squares-Discriminant Analysis (PLS-DA) to assess the classification performance of the microbiota with indicators expressed in classes. Results This study offers four key insights. Firstly, among all indicators, two effectively captured resilience. Secondly, our analyses revealed robust relationship between microbial composition and resilience in terms of both composition and richness. We found decreased α-diversity in less-resilient animals, while specific amplicon sequence variants (ASVs) and KEGG pathways associated with inflammatory responses were negatively linked to resilience. Thirdly, considering resilience indicators in classes, we observed significant differences in microbial composition primarily in animals with lower resilience. Lastly, our study indicates that gut microbial composition can serve as a reliable biomarker for distinguishing individuals with lower resilience. Conclusion Our comprehensive analyses have highlighted the host-microbiota and resilience connection, contributing valuable insights to the existing scientific knowledge. The practical implications of PLS-DA and microbiability results are noteworthy. PLS-DA suggests that host-microbiota interactions could be utilized as biomarkers for monitoring resilience. Furthermore, the microbiability findings show that leveraging host-microbiota insights may improve the identification of resilient animals, supporting their adaptive capacity in response to changing environmental conditions. These practical implications offer promising avenues for enhancing animal well-being and adaptation strategies in the context of environmental challenges faced by livestock populations. Video Abstract

Abstract Antisense RNAs (asRNAs) represent an underappreciated yet crucial layer of gene expression regulation. Generally thought to modulate their sense genes in cis through sequence complementarity or their act of transcription, asRNAs can also regulate different molecular targets in trans, in the nucleus or in the cytoplasm. Here, we performed an in-depth molecular characterization of NFYC Antisense 1 (NFYC-AS1), the asRNA transcribed head-to-head to NFYC subunit of the proliferation-associated NF-Y transcription factor. Our results show that NFYC-AS1 is a prevalently nuclear asRNA peaking early in the cell cycle. Comparative genomics suggests a narrow phylogenetic distribution, with a probable origin in the common ancestor of mammalian lineages. NFYC-AS1 is overexpressed pancancer, preferentially in association with RB1 mutations. Knockdown of NFYC-AS1 by antisense oligonucleotides impairs cell growth in lung squamous cell carcinoma and small cell lung cancer cells, a phenotype recapitulated by CRISPR/Cas9-deletion of its transcription start site. Surprisingly, expression of the sense gene is affected only when endogenous transcription of NFYC-AS1 is manipulated. This suggests that regulation of cell proliferation is at least in part independent of the in cis transcription-mediated effect on NFYC and is possibly exerted by RNA-dependent in trans effects converging on the regulation of G2/M cell cycle phase genes. Accordingly, NFYC-AS1-depleted cells are stuck in mitosis, indicating defects in mitotic progression. Overall, NFYC-AS1 emerged as a cell cycle-regulating asRNA with dual action, holding therapeutic potential in different cancer types, including the very aggressive RB1-mutated tumors.

Abstract Background Within the same species, individuals show marked variation in their social dominance. Studies on a handful of populations have indicated heritable genetic variation for this trait, which is determined by both the genetic background of the individual (direct genetic effect) and of its opponent (indirect genetic effect). However, the evolutionary consequences of selection for this trait are largely speculative, as it is not a usual target of selection in livestock populations. Moreover, studying social dominance presents the challenge of working with a phenotype with a mean value that cannot change in the population, as for every winner of an agonistic interaction there will necessarily be a loser. Thus, to investigate what could be the evolutionary response to selection for social dominance, it is necessary to focus on traits that might be correlated with it. This study investigated the genetic correlations of social dominance, both direct and indirect, with several morphology and fitness traits. We used a dataset of agonistic contests involving cattle (Bos taurus): during these contests, pairs of cows compete in ritualized interactions to assess social dominance. The outcomes of 37,996 dominance interactions performed by 8789 cows over 20 years were combined with individual data for fertility, mammary health, milk yield and morphology and analysed using bivariate animal models including indirect genetic effects. Results We found that winning agonistic interactions has a positive genetic correlation with more developed frontal muscle mass, lower fertility, and poorer udder health. We also discovered that the trends of changes in the estimated breeding values of social dominance, udder health and more developed muscle mass were consistent with selection for social dominance in the population. Conclusions We present evidence that social dominance is genetically correlated with fitness traits, as well as empirical evidence of the possible evolutionary trade-offs between these traits. We show that it is feasible to estimate genetic correlations involving dyadic social traits.

We evaluated SARS-CoV-2 antibody response in voluntary blood donors in Italy at different timepoints. Immediately after lockdown easing, 908/25,657 donors (3.5%) had low IgG titers against nucleocapsid. In the next 2 years, titers increased despite few COVID-19 symptoms. On multivariate analysis, allergic rhinitis was associated with reduced risk for symptomatic COVID-19.

Abstract NF-Y is a trimeric transcription factor whose binding site -the CCAAT box- is enriched in cancer-promoting genes. The regulatory subunit, the sequence-specificity conferring NF-YA, comes in two major isoforms, NF-YA long (NF-YAl) and short (NF-YAs). Extensive expression analysis in epithelial cancers determined two features: widespread overexpression and changes in NF-YAl/NF-YAs ratios (NF-YAr) in tumours with EMT features. We performed wet and in silico experiments to explore the role of the isoforms in breast -BRCA- and gastric -STAD- cancers. We generated clones of two Claudinlow BRCA lines SUM159PT and BT549 ablated of exon-3, thus shifting expression from NF-YAl to NF-YAs. Edited clones show normal growth but reduced migratory capacities in vitro and ability to metastatize in vivo. Using TCGA, including upon deconvolution of scRNA-seq data, we formalize the clinical importance of high NF-YAr, associated to EMT genes and cell populations. We derive a novel, prognostic 158 genes signature common to BRCA and STAD Claudinlow tumours. Finally, we identify splicing factors associated to high NF-YAr, validating RBFOX2 as promoting expression of NF-YAl. These data bring three relevant results: (i) the definition and clinical implications of NF-YAr and the 158 genes signature in Claudinlow tumours; (ii) genetic evidence of 28 amino acids in NF-YAl with EMT-promoting capacity; (iii) the definition of selected splicing factors associated to NF-YA isoforms.

ABSTRACT: Reggiana and Modenese are autochthonous cattle breeds, reared in the North of Italy, that can be mainly distinguished for their standard coat color (Reggiana is red, whereas Modenese is white with some pale gray shades). Almost all milk produced by these breeds is transformed into 2 mono-breed branded Parmigiano-Reggiano cheeses, from which farmers receive the economic incomes needed for the sustainable conservation of these animal genetic resources. After the setting up of their herd books in 1960s, these breeds experienced a strong reduction in the population size that was subsequently reverted starting in the 1990s (Reggiana) or more recently (Modenese) reaching at present a total of about 2,800 and 500 registered cows, respectively. Due to the small population size of these breeds, inbreeding is a very important cause of concern for their conservation programs. Inbreeding is traditionally estimated using pedigree data, which are summarized in an inbreeding coefficient calculated at the individual level (FPED). However, incompleteness of pedigree information and registration errors can affect the effectiveness of conservation strategies. High-throughput SNP genotyping platforms allow investigation of inbreeding using genome information that can overcome the limits of pedigree data. Several approaches have been proposed to estimate genomic inbreeding, with the use of runs of homozygosity (ROH) considered to be the more appropriate. In this study, several pedigree and genomic inbreeding parameters, calculated using the whole herd book populations or considering genotyping information (GeneSeek GGP Bovine 150K) from 1,684 Reggiana cattle and 323 Modenese cattle, were compared. Average inbreeding values per year were used to calculate effective population size. Reggiana breed had generally lower genomic inbreeding values than Modenese breed. The low correlation between pedigree-based and genomic-based parameters (ranging from 0.187 to 0.195 and 0.319 to 0.323 in the Reggiana and Modenese breeds, respectively) reflected the common problems of local populations in which pedigree records are not complete. The high proportion of short ROH over the total number of ROH indicates no major recent inbreeding events in both breeds. ROH islands spread over the genome of the 2 breeds (15 in Reggiana and 14 in Modenese) identified several signatures of selection. Some of these included genes affecting milk production traits, stature, body conformation traits (with a main ROH island in both breeds on BTA6 containing the ABCG2, NCAPG, and LCORL genes) and coat color (on BTA13 in Modenese containing the ASIP gene). In conclusion, this work provides an extensive comparative analysis of pedigree and genomic inbreeding parameters and relevant genomic information that will be useful in the conservation strategies of these 2 iconic local cattle breeds.

Introduction: The Italian peninsula is in the center of the Mediterranean area, and historically it has been a hub for numerous human populations, cultures, and also animal species that enriched the hosted biodiversity. Horses are no exception to this phenomenon, with the peculiarity that the gene pool has been impacted by warfare and subsequent “colonization”. In this study, using a comprehensive dataset for almost the entire Italian equine population, in addition to the most influential cosmopolitan breeds, we describe the current status of the modern Italian gene pool.Materials and Methods: The Italian dataset comprised 1,308 individuals and 22 breeds genotyped at a 70 k density that was merged with publicly available data to facilitate comparison with the global equine diversity. After quality control and supervised subsampling to ensure consistency among breeds, the merged dataset with the global equine diversity contained data for 1,333 individuals from 54 populations. Multidimensional scaling, admixture, gene flow, and effective population size were analyzed.Results and Discussion: The results show that some of the native Italian breeds preserve distinct gene pools, potentially because of adaptation to the different geographical contexts of the peninsula. Nevertheless, the comparison with international breeds highlights the presence of strong gene flow from renowned breeds into several Italian breeds, probably due to historical introgression. Coldblood breeds with stronger genetic identity were indeed well differentiated from warmblood breeds, which are highly admixed. Other breeds showed further peculiarities due to their breeding history. Finally, we observed some breeds that exist more on cultural, traditional, and geographical point of view than due to actual genetic distinctiveness.

Abstract NF-Y is a pioneer transcription factor—TF—formed by the Histone-like NF-YB/NF-YC subunits and the regulatory NF-YA. It binds to the CCAAT box, an element enriched in promoters of genes overexpressed in many types of cancer. NF-YA is present in two major isoforms—NF-YAs and NF-YAl—due to alternative splicing, overexpressed in epithelial tumors. Here we analyzed NF-Y expression in stomach adenocarcinomas (STAD). We completed the partitioning of all TCGA tumor samples (450) according to molecular subtypes proposed by TCGA and ACRG, using the deep learning tool DeepCC. We analyzed differentially expressed genes—DEG—for enriched pathways and TFs binding sites in promoters. CCAAT is the predominant element only in the core group of genes upregulated in all subtypes, with cell-cycle gene signatures. NF-Y subunits are overexpressed, particularly NF-YA. NF-YAs is predominant in CIN, MSI and EBV TCGA subtypes, NF-YAl is higher in GS and in the ACRG EMT subtypes. Moreover, NF-YAlhigh tumors correlate with a discrete Claudinlow cohort. Elevated NF-YB levels are protective in MSS;TP53+ patients, whereas high NF-YAl/NF-YAs ratios correlate with worse prognosis. We conclude that NF-Y isoforms are associated to clinically relevant features of gastric cancer.

Satellite cells represent myogenic stem cells that allow the homeostasis and repair of adult skeletal muscle. Here the authors report that the transcription factor NF-Y is expressed in satellite cells and is important for their maintenance and proper myogenic differentiation.

Complex genetic and phenotypic relationships are theorized to link different fitness components but revealing the correlations occurring among disparate traits requires large datasets of pedigreed populations. In particular, the association between traits beneficial to social dominance with health and fitness could be antagonistic, because of trade-offs, or positive, because of greater resource acquisition by dominant individuals. Studies investigating these relationships found some empirical evidence in support of both theories, mainly using multiple trait models (MTM). However, if a trait giving a social advantage is suspected to affect the expression of other traits, MTM could provide some bias, that structural equation models (SEM) could highlight. We used Aosta Chestnut-Black Pied cattle to investigate whether the fighting ability of cows (the capability of winning social dominance interactions) is genetically correlated with health and fitness traits. We ran both MTM and SEM using a Gibbs sampling algorithm to disentangle the possible causal effects of fighting ability from the genetic correlations that this trait shares with other traits: individual milk yield, somatic cells (representing mammary health), fertility, and longevity. We found antagonistic genetic correlations, similar under both approaches, for fighting ability vs. milk, somatic cells, and fertility, Accordingly, we found only a slight causal effects of fighting ability on these traits (–0.012 to 0.059 in standardized value). However, we found genetic correlations opposite in sign between fighting ability and longevity under MTM (0.237) and SEM (–0.183), suggesting a strong causal effect (0.386 standardized) of fighting ability on longevity. In other words, MTM found a positive correlation between longevity and fighting ability, while SEM found a negative correlation. The explanation could be that for economic reasons dominant cows are kept in this population for longer, thus attaining greater longevity: using MTM, the economic importance of competitions probably covers the true genetic correlation among traits. This artificially simulates a natural situation where an antagonistic genetic correlation between longevity and fighting ability appears positive under MTM due to a non-genetic advantage obtained by the best fighters. The use of SEM to properly assess the relationships among traits is suggested in both evolutionary studies and animal breeding.

Abstract Transcription factors (TFs) are core players in the control of gene expression, evolutionarily selected to recognise a subset of specific DNA sequences and nucleate the recruitment of the transcriptional machinery. How TFs assemble and move in the nucleus to locate and bind their DNA targets and cause a transcriptional response, remains mostly unclear. NF-Y is a highly conserved, heterotrimeric TF with important roles in both housekeeping and lineage-specific gene expression, functioning as a promoter organiser. Despite a large number of biochemical, structural and genomic studies of NF-Y, there is a lack of experiments in single living cells; therefore, basic assumptions of NF-Y biology remain unproven in vivo. Here we employ a series of dynamic fluorescence microscopy methods (FLIM-FRET, NB, RICS and FRAP) to study NF-Y dynamics and complex formation in live cells. Specifically, we provide quantitative measurement of NF-Y subunit association and diffusion kinetics in the nucleus that collectively suggest NF-Y to move and bind chromatin as a trimeric complex in vivo.

Genomic selection has been increasingly implemented in the animal breeding industry, and it is becoming a routine method in many livestock breeding contexts. However, its use is still limited in several small-population local breeds, which are, nonetheless, an important source of genetic variability of great economic value. A major roadblock for their genomic selection is accuracy when population size is limited: to improve breeding value accuracy, variable selection models that assume heterogenous variance have been proposed over the last few years. However, while these models might outperform traditional and genomic predictions in terms of accuracy, they also carry a proportional increase of breeding value bias and dispersion. These mutual increases are especially striking when genomic selection is performed with a low number of phenotypes and high shrinkage value—which is precisely the situation that happens with small local breeds. In our study, we tested several alternative methods to improve the accuracy of genomic selection in a small population. First, we investigated the impact of using only a subset of informative markers regarding prediction accuracy, bias, and dispersion. We used different algorithms to select them, such as recursive feature eliminations, penalized regression, and XGBoost. We compared our results with the predictions of pedigree-based BLUP, single-step genomic BLUP, and weighted single-step genomic BLUP in different simulated populations obtained by combining various parameters in terms of number of QTLs and effective population size. We also investigated these approaches on a real data set belonging to the small local Rendena breed. Our results show that the accuracy of GBLUP in small-sized populations increased when performed with SNPs selected via variable selection methods both in simulated and real data sets. In addition, the use of variable selection models—especially those using XGBoost—in our real data set did not impact bias and the dispersion of estimated breeding values. We have discussed possible explanations for our results and how our study can help estimate breeding values for future genomic selection in small breeds.

Abstract Background Assessment of genetic diversity and population structure provides important control metrics to avoid genetic erosion, inbreeding depression and crossbreeding between exotic and locally-adapted cattle breeds since these events can have deleterious consequences and eventually lead to extinction. Historically, the Alpine Arc represents an important pocket of cattle biodiversity with a large number of autochthonous breeds that provide a fundamental source of income for the entire regional economy. By using genotype data from medium-density single nucleotide polymorphism (SNP) arrays, we performed a genome-wide comparative study of 23 cattle populations from the Alpine Arc and three cosmopolitan breeds. Results After filtering, we obtained a final genotyping dataset consisting of 30,176 SNPs for 711 individuals. The local breeds showed high or intermediate values of genetic diversity compared to the highly selected cosmopolitan breeds. Patterns of genetic differentiation, multidimensional scaling, admixture analysis and the constructed phylogenetic tree showed convergence, which indicates the presence of gene flow among the breeds according to both geographic origin and historical background. Among the most differentiated breeds, we identified the modern Brown cattle. In spite of admixture events, several local breeds have preserved distinctive characteristics, which is probably due to differences in genetic origin and geographic location. Conclusions This study represents one of the most comprehensive genome-wide analysis of the Alpine cattle breeds to date. Using such a large dataset that includes the majority of the local breeds found in this region, allowed us to expand knowledge on the evaluation and status of Alpine cattle biodiversity. Our results indicate that although many of the analyzed local breeds are listed as endangered, they still harbor a large amount of genetic diversity, even when compared to some cosmopolitan breeds. This finding, together with the reconstruction of the phylogeny and the relationships between these Alpine Arc cattle breeds, provide crucial insights not only into the improvement of genetic stocks but also into the implementation of future conservation strategies.

Knowledge of the genetic architecture of key growth and beef traits in livestock species has greatly improved worldwide thanks to genome-wide association studies (GWAS), which allow to link target phenotypes to Single Nucleotide Polymorphisms (SNPs) across the genome. Local dual-purpose breeds have rarely been the focus of such studies; recently, however, their value as a possible alternative to intensively farmed breeds has become clear, especially for their greater adaptability to environmental change and potential for survival in less productive areas. We performed single-step GWAS and post-GWAS analysis for body weight (BW), average daily gain (ADG), carcass fleshiness (CF) and dressing percentage (DP) in 1,690 individuals of local alpine cattle breed, Rendena. This breed is typical of alpine pastures, with a marked dual-purpose attitude and good genetic diversity. Moreover, we considered two of the target phenotypes (BW and ADG) at different times in the individuals’ life, a potentially important aspect in the study of the traits’ genetic architecture. We identified 8 significant and 47 suggestively associated SNPs, located in 14 autosomal chromosomes (BTA). Among the strongest signals, 3 significant and 16 suggestive SNPs were associated with ADG and were located on BTA10 (50–60 Mb), while the hotspot associated with CF and DP was on BTA18 (55–62 MB). Among the significant SNPs some were mapped within genes, such as SLC12A1, CGNL1, PRTG (ADG), LOC513941 (CF), NLRP2 (CF and DP), CDC155 (DP). Pathway analysis showed great diversity in the biological pathways linked to the different traits; several were associated with neurogenesis and synaptic transmission, but actin-related and transmembrane transport pathways were also represented. Time-stratification highlighted how the genetic architectures of the same traits were markedly different between different ages. The results from our GWAS of beef traits in Rendena led to the detection of a variety of genes both well-known and novel. We argue that our results show that expanding genomic research to local breeds can reveal hitherto undetected genetic architectures in livestock worldwide. This could greatly help efforts to map genomic complexity of the traits of interest and to make appropriate breeding decisions.

Preliminary the figure of Abbot Feliciano Scarpellini and the scientific environment in Rome in the first half of the XIX century are introduced. It is then analysed the life and work of the physicist and philosopher Giambattista Pianciani (1784-1862) of the Society of Jesus. Pianciani was in Rome among the most peculiar representatives of that cultural program aimed at countering the theses of Enlightenment materialism through a process of conciliation and justification of scientific discoveries with Catholic doctrine. This apologetic and concordist program started under Pius VII, also continued during the pontificate of Leo XVI. In the wake of this peculiar apologetic program, the controversy that, around the mid-nineteenth century, arose in the Roman College between neotomists and some scientists of the order, for example, Angelo Secchi, is also discussed. Finally, the specific concordist program developed by Pianciani is discussed. This program aimed to reconcile the biblical account of the world's creation with the scientific results that came from the studies of natural cosmogony.

In its 800-year history, the University of Padova (Padua, Italy) has come to play an important role in the development of animal science in Italy and Europe. Having founded the oldest university botanical garden (1545; UNESCO World Heritage Site) and anatomical theatre (1595), and awarded the first university degree to a woman (Elena-Lucrezia Cornaro-Piscopia, 1678), the University instituted the Public School of Agriculture (Orto Agrario) in 1765 and the Collegium Zooiatricum in 1773. Between 1986 and 1995, under the leadership of Professor Mario Bonsembiante, animal scientist and rector, the scientific-technological center of Agripolis was established, including the Faculties of Agriculture and Veterinary Medicine (with the experimental farm and veterinary hospital), the Istituto Zooprofilattico delle Venezie, and Veneto Agricoltura. Agripolis rapidly became the most innovative center in Italy for teaching, research, and extension services in the fields of agricultural, animal, veterinary, food and environmental sciences. In the last 35 years, the number of Italian animal scientists has remained almost constant, but they have moved on to more innovative research topics closer to society’s demand and concerns, and significantly enhanced their international standing, reaching the first position in Europe and the fourth in the world in terms of citations.

Population structure or genetic relatedness should be considered in genome association studies to avoid spurious association. The most used methods for genome-wide association studies (GWAS) account for population structure but are limited to genotyped individuals with phenotypes. Single-step GWAS (ssGWAS) can use phenotypes from non-genotyped relatives; however, its ability to account for population structure has not been explored. Here we investigate the equivalence among ssGWAS, efficient mixed-model association expedited (EMMAX), and genomic best linear unbiased prediction GWAS (GBLUP-GWAS), and how they differ from the single-SNP analysis without correction for population structure (SSA-NoCor). We used simulated, structured populations that mimicked fish, beef cattle, and dairy cattle populations with 1040, 5525, and 1,400 genotyped individuals, respectively. Larger populations were also simulated that had up to 10-fold more genotyped animals. The genomes were composed by 29 chromosomes, each harboring one QTN, and the number of simulated SNPs was 35,000 for the fish and 65,000 for the beef and dairy cattle populations. Males and females were genotyped in the fish and beef cattle populations, whereas only males had genotypes in the dairy population. Phenotypes for a trait with heritability varying from 0.25 to 0.35 were available in both sexes for the fish population, but only for females in the beef and dairy cattle populations. In the latter, phenotypes of daughters were projected into genotyped sires (i.e., deregressed proofs) before applying EMMAX and SSA-NoCor. Although SSA-NoCor had the largest number of true positive SNPs among the four methods, the number of false negatives was two–fivefold that of true positives. GBLUP-GWAS and EMMAX had a similar number of true positives, which was slightly smaller than in ssGWAS, although the difference was not significant. Additionally, no significant differences were observed when deregressed proofs were used as pseudo-phenotypes in EMMAX compared to daughter phenotypes in ssGWAS for the dairy cattle population. Single-step GWAS accounts for population structure and is a straightforward method for association analysis when only a fraction of the population is genotyped and/or when phenotypes are available on non-genotyped relatives.

NF-Y is a trimeric Transcription Factor -TF- which binds with high selectivity to the conserved CCAAT element. Individual ChIP-seq analysis as well as ENCODE have progressively identified locations shared by other TFs. Here, we have analyzed data introduced by ENCODE over the last five years in K562, HeLa-S3 and GM12878, including several chromatin features, as well RNA-seq profiling of HeLa cells after NF-Y inactivation. We double the number of sequence-specific TFs and co-factors reported. We catalogue them in 4 classes based on co-association criteria, infer target genes categorizations, identify positional bias of binding sites and gene expression changes. Larger and novel co-associations emerge, specifically concerning subunits of repressive complexes as well as RNA-binding proteins. On the one hand, these data better define NF-Y association with single members of major classes of TFs, on the other, they suggest that it might have a wider role in the control of mRNA production.

Local breeds are often reared in various environmental conditions (EC), suggesting that genotype by environment interaction (GxE) could influence genetic progress. This study aimed at investigating GxE and response to selection (R) in Rendena cattle under diverse EC. Traits included milk, fat, and protein yields, fat and protein percentage, and somatic cell score, three-factor scores and 24 linear type traits. The traits belonged to 11,085 cows (615 sires). Variance components were estimated in a two-step reaction norm model (RNM). A single trait animal model was run to obtain the solutions of herd-EC effect, then included in a random regression sire model. A multivariate response to selection (R) in different EC was computed for traits under selection including beef traits from a performance test. GxE accounted on average for 10% of phenotypic variance, and an average rank correlation of over 0.97 was found between bull estimated breeding values (EBVs) by either including or not including GxE, with changing top ranks. For various traits, significantly greater genetic components and R were observed in plain farms, loose housing rearing system, feeding total mixed ration, and without summer pasture. Conversely, for beef traits, a greater R was found for mountain farms, loose housing, hay-based feeding and summer pasture.

Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-Seq) has opened new avenues of research in the genome-wide characterization of regulatory DNA-protein interactions at the genetic and epigenetic level. As a consequence, it has become the de facto standard for studies on the regulation of transcription, and literally thousands of data sets for transcription factors and cofactors in different conditions and species are now available to the scientific community. However, while pipelines and best practices have been established for the analysis of a single experiment, there is still no consensus on the best way to perform an integrated analysis of multiple datasets in the same condition, in order to identify the most relevant and widespread regulatory modules composed by different transcription factors and cofactors. We present here a computational pipeline for this task, that integrates peak summit colocalization, a novel statistical framework for the evaluation of its significance, and motif enrichment analysis. We show examples of its application to ENCODE data, that led to the identification of relevant regulatory modules composed of different factors, as well as the organization on DNA of the binding motifs responsible for their recruitment.

Abstract Background In the last 50 years, the diversity of cattle breeds has experienced a severe contraction. However, in spite of the growing diffusion of cosmopolite specialized breeds, several local cattle breeds are still farmed in Italy. Genetic characterization of breeds represents an essential step to guide decisions in the management of farm animal genetic resources. The aim of this work was to provide a high-resolution representation of the genome-wide diversity and population structure of Italian local cattle breeds using a medium-density single nucleotide polymorphism (SNP) array. Results After quality control filtering, the dataset included 31,013 SNPs for 800 samples from 32 breeds. Our results on the genetic diversity of these breeds agree largely with their recorded history. We observed a low level of genetic diversity, which together with the small size of the effective populations, confirmed that several breeds are threatened with extinction. According to the analysis of runs of homozygosity, evidence of recent inbreeding was strong in some local breeds, such as Garfagnina, Mucca Pisana and Pontremolese. Patterns of genetic differentiation, shared ancestry, admixture events, and the phylogenetic tree, all suggest the presence of gene flow, in particular among breeds that originate from the same geographical area, such as the Sicilian breeds. In spite of the complex admixture events that most Italian cattle breeds have experienced, they have preserved distinctive characteristics and can be clearly discriminated, which is probably due to differences in genetic origin, environment, genetic isolation and inbreeding. Conclusions This study is the first exhaustive genome-wide analysis of the diversity of Italian cattle breeds. The results are of significant importance because they will help design and implement conservation strategies. Indeed, efforts to maintain genetic diversity in these breeds are needed. Improvement of systems to record and monitor inbreeding in these breeds may contribute to their in situ conservation and, in view of this, the availability of genomic data is a fundamental resource.

The maintenance of local cattle breeds is key to selecting for efficient food production, landscape protection, and conservation of biodiversity and local cultural heritage. Rendena is an indigenous cattle breed from the alpine North-East of Italy, selected for dual purpose, but with lesser emphasis given to beef traits. In this situation, increasing accuracy for beef traits could prevent detrimental effects due to the antagonism with milk production. Our study assessed the impact of genomic information on estimated breeding values (EBVs) in Rendena performance-tested bulls. Traits considered were average daily gain, in vivo EUROP score, and in vivo estimate of dressing percentage. The final dataset contained 1691 individuals with phenotypes and 8372 animals in pedigree, 1743 of which were genotyped. Using the cross-validation method, three models were compared: (i) Pedigree-BLUP (PBLUP); (ii) single-step GBLUP (ssGBLUP), and (iii) weighted single-step GBLUP (WssGBLUP). Models including genomic information presented higher accuracy, especially WssGBLUP. However, the model with the best overall properties was the ssGBLUP, showing higher accuracy than PBLUP and optimal values of bias and dispersion parameters. Our study demonstrated that integrating phenotypes for beef traits with genomic data can be helpful to estimate EBVs, even in a small local breed.

Selection in local dual-purpose breeds requires great carefulness because of the need to preserve peculiar traits and also guarantee the positive genetic progress for milk and beef production to maintain economic competitiveness. A specific breeding plan accounting for milk, beef, and functional traits is required by breeders of the Alpine Grey cattle (AG), a local dual-purpose breed of the Italian Alps. Hereditability and genetic correlations among all traits have been analyzed for this purpose. After that, different selection indexes were proposed to identify the most suitable for this breed. Firstly, a genetic parameters analysis was carried out with different datasets. The milk dataset contained 406,918 test day records of milk, protein, and fat yields and somatic cells (expressed as SCS). The beef dataset included performance test data conducted on 749 young bulls. Average daily gain, in vivo estimated carcass yields, and carcass conformation (SEUROP) were the phenotypes obtained from the performance tests. The morphological dataset included 21 linear type evaluations of 11,320 first party cows. Linear type traits were aggregated through factor analysis and three factors were retained, while head typicality (HT) and rear muscularity (RM) were analyzed as single traits. Heritability estimates (h2) for milk traits ranged from 0.125 to 0.219. Analysis of beef traits showed h2 greater than milk traits, ranging from 0.282 to 0.501. Type traits showed a medium value of h2 ranging from 0.238 to 0.374. Regarding genetic correlation, SCS and milk traits were strongly positively correlated. Milk traits had a negative genetic correlation with the factor accounting for udder conformations (−0.40) and with all performance test traits and RM. These latter traits showed also a negative genetic correlation with udder volume (−0.28). The HT and the factor accounting for rear legs traits were not correlated with milk traits, but negatively correlated with beef traits (−0.32 with RM). We argue that the consequence of these results is that the use of the current selection index, which is mainly focused on milk attitude, will lead to a deterioration of all other traits. In this study, we propose more appropriate selection indexes that account for genetic relationships among traits, including functional traits.

NF-Y is a pioneer trimeric transcription factor formed by the Histone Fold Domain (HFD) NF-YB/NF-YC subunits and NF-YA. Three subunits are required for DNA binding. CCAAT-specificity resides in NF-YA and transactivation resides in Q-rich domains of NF-YA and NF-YC. They are involved in alternative splicing (AS). We recently showed that NF-YA is overexpressed in breast and lung carcinomas. We report here on the overexpression of all subunits in the liver hepatocellular carcinoma (HCC) TCGA database, specifically the short NF-YAs and NF-YC2 (37 kDa) isoforms. This is observed at all tumor stages, in viral-infected samples and independently from the inflammatory status. Up-regulation of NF-YAs and NF-YC, but not NF-YB, is associated to tumors with mutant p53. We used a deep-learning-based method (DeepCC) to extend the partitioning of the three molecular clusters to all HCC TCGA tumors. In iCluster3, CCAAT is a primary matrix found in promoters of up-regulated genes, and cell-cycle pathways are enriched. Finally, clinical data indicate that, globally, only NF-YAs, but not HFD subunits, correlate with the worst prognosis; in iCluster1 patients, however, all subunits correlate. The data show a difference with other epithelial cancers, in that global overexpression of the three subunits is reported and clinically relevant in a subset of patients; yet, they further reinstate the regulatory role of the sequence-specific subunit.

The Aosta Red Pied (Valdostana Pezzata Rossa (VRP)), the Aosta Black Pied (Valdostana Pezzata Nera (VBP)) and the Aosta Chestnut (Valdostana Castana (CAS)) are dual-purpose cattle breeds (meat and milk), very well adapted to the harsh environmental conditions of alpine territories: their farming is in fact characterized by summer pasture at very high altitude. A total of 728 individuals were genotyped with the GeenSeek Genomic Profiler® (GGP) Bovine 150K Illumina SNP chip as a part of the DUALBREEDING-PSRN Italian-funded research project. The genetic diversity among populations showed that the three breeds are distinct populations based on the FST values, ADMIXTURE and Principal Component Analysis (PCA) results. Runs of Homozygosity (ROH) were obtained for the three populations to disclose recent autozygosity. The genomic inbreeding based on the ROH was calculated and coupled with information derived from the F (inbreeding coefficient) and FST parameters. The mean FROH values were low: CAS = 0.06, VBP = 0.05 and VRP = 0.07, while the average F values were −0.003, −0.01 and −0.003, respectively. The annotation and enrichment analysis, performed in the identified most frequent ROH (TOP_ROH), showed genes that can be linked to the resilience capacity of these populations to harsh environmental farming conditions, and to the peculiar characteristics searched for by farmers in each breed.

Various supplementations in animal feeding have been investigate in order to enrich food of animal origin with n-3 fatty acids. Although the effects of flaxseeds inclusion in diets for lactating dairy have already been assessed, few studies have focused on this n-3 source supplementation during the transition period. The aim of this work was to evaluate the effects of flacked flaxseed (200 g/head/day; 2.13% DM) dietary treatment during the dry period on milk yield and quality in the 30 days after calving. In addition, the enterolactone content in plasma (before and after calving) and in milk of cows fed diets supplemented or not with flaxseed was considered. The study demonstrated that the carry-over effect on the milk profile of C18:2, C18:3 n-3, and C20:5 n-3 was significantly higher in flaxseed diet than in the control one at 4th day of lactation. A significant increase of enterolactone on milk from flaxseed fed cows was observed only at 15 sampling day. The quick modification in fatty acid (FA) profile of the milk in the first few days of lactation suggests that the carry over effect from pre-calving flaxseed feeding at this concentration was very short lasting.

Several studies reveal that coenzyme Q (CoQ) and vitamin E (Vit. E) act against oxidative deterioration, and that CoQ restores the active and antioxidant form of Vit. E. These two antioxidants, acting against lipid peroxidation, seem to be able to improve motility parameters of spermatozoa. The objective of this study is to evaluate the addition of CoQ and Vit. E to semen extender for equine spermatozoa in order to evaluate possible effects on semen motility. First, immediately after collection, semen samples were diluted with 1mM of CoQ and 1mM of CoQ plus 1mM of Vit. E and prepared for frozen storage in liquid nitrogen. After thawing (37 °C/30 s), samples were maintained at 37 °C and subjected to analysis after 0, 2 and 4 h for motility parameters with CASA (Computer-Assisted Sperm Analysis) method. In a second experiment, after the collection, semen samples were diluted with 1mM of CoQ, in presence or absence of seminal plasma where Vit. E is normally present, and prepared for cooled storage at 4 °C. The effects on motility parameters were determined with CASA at 0, 24, 31 and 48 h after collection. During the analysis, samples were kept at 4 °C. The CASA variables were examined with a mixed linear model. No improvement (p > .05) in motility parameters results from the addition of CoQ and Vit. E in frozen or cooled-stored equine semen when compared to control group.

The recovery of spermatozoa from the cauda epididymis may be the last chance to obtain genetic material from stallions undergone to castration. The aim of the current study was to evaluate the efficiency of cryopreservation and the use of two different extenders for equine epididymal spermatozoa. Testicles obtained from castration were divided into two groups: cauda epididymis processed immediately after orchiectomy and cauda epididymis processed after 24 h storage in saline solution at 4 °C of the testis. The epididymal spermatozoa were collected through manual slicing of the cauda epididymis of each testicle. In addition, spermatozoa obtained from different processed testes were diluted alternatively with either modified Palmer or EGG TECH® extenders to produce frozen straws. Motility parameters in fresh and frozen-thawed material were analysed by means of the computer-aided sperm analysis (CASA). The recorded CASA data were analysed with a mixed linear model. Motility parameters in fresh semen yielded better results than in frozen semen (p = 0.008), but no difference (p > 0.05) was observed between spermatozoa collected immediately after castration or after 24 h of storage; in frozen-thawed samples, EGG TECH® tended to improve the percentage of progressively motile spermatozoa in epididymal frozen-thawed semen (p = 0.08) compared with modified Palmer. We conclude that the processing of epididymal spermatozoa can occur up to 24 h after stallion castration and both common extenders used are suitable for preserving this material.

NF-Y is a transcription factor (TF) comprising three subunits (NF-YA, NF-YB, NF-YC) that binds with high specificity to the CCAAT sequence, a widespread regulatory element in gene promoters of prosurvival, cell-cycle-promoting, and metabolic genes. Tumor cells undergo “metabolic rewiring” through overexpression of genes involved in such pathways, many of which are under NF-Y control. In addition, NF-YA appears to be overexpressed in many tumor types. Thus, limiting NF-Y activity may represent a desirable anti-cancer strategy, which is an ongoing field of research. With virtual-screening docking simulations on a library of pharmacologically active compounds, we identified suramin as a potential NF-Y inhibitor. We focused on suramin given its high water-solubility that is an important factor for in vitro testing, since NF-Y is sensitive to DMSO. By electrophoretic mobility shift assays (EMSA), isothermal titration calorimetry (ITC), STD NMR, X-ray crystallography, and molecular dynamics (MD) simulations, we showed that suramin binds to the histone fold domains (HFDs) of NF-Y, preventing DNA-binding. Our analyses, provide atomic-level detail on the interaction between suramin and NF-Y and reveal a region of the protein, nearby the suramin-binding site and poorly conserved in other HFD-containing TFs, that may represent a promising starting point for rational design of more specific and potent inhibitors with potential therapeutic applications.

This study aimed to investigate the genetic diversity in the Italian Heavy Horse Breed from pedigree and genomic data. Pedigree information for 64,917 individuals were used to assess inbreeding level, effective population size (Ne), and effective numbers of founders and ancestors (fa/fe). Genotypic information from SNP markers were available for 267 individuals of both sexes, and it allowed estimating genomic inbreeding in two methods (observed versus expected homozygosity and from ROH) to study the breed genomic structure and possible selection signatures. Pedigree and genomic inbreeding were greatly correlated (0.65 on average). The inbreeding trend increased over time, apart from periods in which the base population enlarged, when Ne increased also. Recent bottlenecks did not occur in the genome, as fa/fe have shown. The observed homozygosity results were on average lower than expected, which was probably due to the use of French Breton stallions to support the breed genetic variability. High homozygous regions suggested that inbreeding increased in different periods. Two subpopulations were distinguished, which was probably due to the different inclusion of French animals by breeders. Few selection signatures were found at the population level, with possible associations to disease resistance. The almost low inbreeding rate suggested that despite the small breed size, conservation actions are not yet required.

The Italian Heavy Draught Horse (IHDH) breed is selected based on linear type traits (LTT) evaluated at young age on six-month-old foals. However, animals retained for reproduction are scored also at adults age (about 30 months), and the evaluation is mandatory for the final official admission to the stud book of candidate mares and stallions. This study aimed to estimate genetic parameters of LTT scored at 30 months to consider if they are feasible for selection instead of using foal data and to reduce costs of selection plan. Data included 19 years of evaluation for 14 LTT and an overall score. Analyses were performed on 5835 females and 856 males via animal model. The heritability ranged from 0.03 (upper line length) to 0.40 (frame size). Traits of selection interest (head size and expression; temperament/movement; fleshiness; fore diameter; rear diameter) reported heritability between 0.21 and 0.31. High genetic correlations were obtained among traits related to muscular development, 0.73 on average. Positive genetic trends were found in traits of selection interest, already selected from foal type trait data. Accounting for genetic parameters estimated in adult animals instead in foals is feasible in IHDH selection.

Our aims were to find a phenotypic variable to express mares’ lifetime reproductive performance after 6 breeding seasons (BS) in Italian Heavy Draught Horse breed (IHDH), and to estimate its heritability. At first, 1487 mares in a training dataset were used to implement and validate a set of predictive coefficients (LFR-C) or equations (LFR-E) to estimate a lifetime foaling rate (LFR) after 6 BS, i.e., the number of foals generated divided by the opportunities to do so. Then, 3033 mares in a dataset with at least 3 registered BS, was used to estimate LFR for mares with 3, 4, or 5 registered RS. This dataset contained actual (n = 1950) and estimated (n = 1443) LFR, obtained by LFR-C, and LFR-E; Arcsine transformation of LFR-C and LFR-E were also analyzed in single trait animal models to estimate heritability. Overall, the LFR showed a moderate but significant genetic variation, and the heritability of the trait was high (0.24) considering it is a fitness trait. The arcsine transformation of LFR did not show any improvement of heritability. The present study indicates the possible use of a linear LFR variable for breeding purposes in IHDH breed considering both complete and incomplete reproductive careers.

NF-YA, the regulatory subunit of the trimeric transcription factor (TF) NF-Y, is regulated by alternative splicing (AS) generating two major isoforms, “long” (NF-YAl) and “short” (NF-YAs). Muscle cells express NF-YAl. We ablated exon 3 in mouse C2C12 cells by a four-guide CRISPR/Cas9n strategy, obtaining clones expressing exclusively NF-YAs (C2-YAl-KO). C2-YAl-KO cells grow normally, but are unable to differentiate. Myogenin and—to a lesser extent, MyoD— levels are substantially lower in C2-YAl-KO, before and after differentiation. Expression of the fusogenic Myomaker and Myomixer genes, crucial for the early phases of the process, is not induced. Myomaker and Myomixer promoters are bound by MyoD and Myogenin, and Myogenin overexpression induces their expression in C2-YAl-KO. NF-Y inactivation reduces MyoD and Myogenin, but not directly: the Myogenin promoter is CCAAT-less, and the canonical CCAAT of the MyoD promoter is not bound by NF-Y in vivo. We propose that NF-YAl, but not NF-YAs, maintains muscle commitment by indirectly regulating Myogenin and MyoD expression in C2C12 cells. These experiments are the first genetic evidence that the two NF-YA isoforms have functionally distinct roles.

Sox3/SOX3 is one of the earliest neural markers in vertebrates. Together with the Sox1/SOX1 and Sox2/SOX2 genes it is implicated in the regulation of stem cell identity. In the present study, we performed the first analysis of epigenetic mechanisms (DNA methylation and histone marks) involved in the regulation of the human SOX3 gene expression during RA-induced neural differentiation of NT2/D1 cells. We show that the promoter of the human SOX3 gene is extremely hypomethylated both in undifferentiated NT2/D1 cells and during the early phases of RA-induced neural differentiation. By employing chromatin immunoprecipitation, we analyze several histone modifications across different regions of the SOX3 gene and their dynamics following initiation of differentiation. In the same timeframe we investigate profiles of selected histone marks on the promoters of human SOX1 and SOX2 genes. We demonstrate differences in histone signatures of SOX1, SOX2 and SOX3 genes. Considering the importance of SOXB1 genes in the process of neural differentiation, the present study contributes to a better understanding of epigenetic mechanisms implicated in the regulation of pluripotency maintenance and commitment towards the neural lineage.

Rice flowering is controlled by changes in the photoperiod that promote the transition to the reproductive phase as days become shorter. Natural genetic variation for flowering time has been largely documented and has been instrumental to define the genetics of the photoperiodic pathway, as well as providing valuable material for artificial selection of varieties better adapted to local environments. We mined genetic variation in a collection of rice varieties highly adapted to European regions and isolated distinct variants of the long day repressor HEADING DATE 1 (Hd1) that perturb its expression or protein function. Specific variants allowed us to define novel features of the photoperiodic flowering pathway. We demonstrate that a histone fold domain scaffold formed by GRAIN YIELD, PLANT HEIGHT AND HEADING DATE 8 (Ghd8) and several NF-YC subunits can accommodate distinct proteins, including Hd1 and PSEUDO RESPONSE REGULATOR 37 (PRR37), and that the resulting OsNF-Y complex containing Hd1 can bind a specific sequence in the promoter of HEADING DATE 3A (Hd3a). Artificial selection has locally favored an Hd1 variant unable to assemble in such heterotrimeric complex. The causal polymorphism was defined as a single conserved lysine in the CCT domain of the Hd1 protein. Our results indicate how genetic variation can be stratified and explored at multiple levels, and how its description can contribute to the molecular understanding of basic developmental processes.

Photoperiod dependent flowering is one of several mechanisms used by plants to initiate the developmental transition from vegetative growth to reproductive growth. The NUCLEAR FACTOR Y (NF-Y) transcription factors are heterotrimeric complexes composed of NF-YA and histone-fold domain (HFD) containing NF-YB/NF-YC, that initiate photoperiod-dependent flowering by cooperatively interacting with CONSTANS (CO) to drive the expression of FLOWERING LOCUS T (FT). This involves NF-Y and CO binding at distal CCAAT and proximal "CORE" elements, respectively, in the FT promoter. While this is well established for the HFD subunits, there remains some question over the potential role of NF-YA as either positive or negative regulators of this process. Here we provide strong support, in the form of genetic and biochemical analyses, that NF-YA, in complex with NF-YB/NF-YC proteins, can directly bind the distal CCAAT box in the FT promoter and are positive regulators of flowering in an FT-dependent manner.

Nuclear Factor Y (NF-Y) is an evolutionarily conserved trimer formed by a Histone-Fold Domain (HFD) heterodimeric module shared by core histones, and the sequence-specific NF-YA subunit. In plants, the genes encoding each of the three subunits have expanded in number, giving rise to hundreds of potential trimers. While in mammals NF-Y binds a well-characterized motif, with a defined matrix centered on the CCAAT box, the specificity of the plant trimers has yet to be determined. Here we report that Arabidopsis thaliana NF-Y trimeric complexes, containing two different NF-YA subunits, bind DNA in vitro with similar affinities. We assayed precisely sequence-specificity by saturation mutagenesis, and analyzed genomic DNA sites bound in vivo by selected HFDs. The plant NF-Y CCAAT matrix is different in nucleotides flanking CCAAT with respect to the mammalian matrix, in vitro and in vivo. Our data point to flexible DNA-binding rules by plant NF-Ys, serving the scope of adapting to a diverse audience of genomic motifs.

Animal DNA barcoding allows researchers to identify different species by analyzing a short nucleotide sequence, typically the mitochondrial gene cox1. In this paper, we use DNA barcoding to genetically identify seafood samples that were purchased from various locations throughout Italy. We adopted a multi-locus approach to analyze the cob, 16S-rDNA and cox1 genes, and compared our sequences to reference sequences in the BOLD and GenBank online databases. Our method is a rapid and robust technique that can be used to genetically identify crustaceans, mollusks and fishes. This approach could be applied in the future for conservation, particularly for monitoring illegal trade of protected and endangered species. Additionally, this method could be used for authentication in order to detect mislabeling of commercially processed seafood.

The objective of this experiment is to evaluate the changes of particle size distribution and chemical composition of the total mixed ration (TMR) based on hay as the main forage component (“dry” TMR) and distributed once (7.00 am) or twice (7.00 am and 1.00 pm) daily to 32 lactating cows. The trial was divided in two periods of 14 days each. Diet (DM=53.7%) was formulated in order to assure the nutritional requirements of cows producing 24 kg/d of milk (crude protein=14.4% DM; NDF=40.9% DM; milk FU=0.88/kg DM) and additional amounts of concentrates were distributed using automatic feeders. Four TMR samples were collected daily (7.00 am, 10.00 am, 1.00 pm, 4.00 pm) for six days during each experiment period for a total number of 48 feed samples. Each feed sample was subjected to the estimation of the particle size distribution using the separator of Pennsylvania State University composed of two sieves (diameters of 19 and 8 mm) and a collector on the bottom, and to the determination of the chemical composition. Changes of all three particle size fractions for TMRs were observed during the day with distributions of the TMR both once and twice daily. With the once daily distribution, the large particles fraction increased linearly (P

The aim of this study was to evaluate the effects of the replacement of about 70% of soybean meal (SBM) with extruded(ES) or toasted (TS) full-fat soybean seeds in diets for lactating cows on milk quality, fatty acid profile, and conjugatedlinoleic acid (CLA) content. Eighteen lactating cows were assigned to 3 groups which received a basal diet, supplementedwith 1.8, 2.1 and 2.1 kg/head, respectively, of SBM, ES and TS. There was no significant effect on milk yield,calculated as the difference between daily yield during the experimental period and the mean of the last 5 days of adaptation(-1.65, -1.29 and -0.20 kg/d, respectively, for SBM, ES and TS; P>0.10) and milk quality parameters (fat, protein,urea and cheese making parameters) among treatments. In the ES group there was a decrease in the short chainFA content (from C4 to C13) in milk fat (9.2 vs 11.0 and 10.8 g/100 g lipids, respectively, for ES, SBM and TS; PMedium chain FA (from C14 to C17) content in milk fat was lower for ES and TS groups compared with SBM (46.8 and48.0 vs 54.8 g/100 g lipids respectively; PSBM group compared to the others (34.3 vs 44.2 and 41.2 g/100 g lipids, respectively, for SBM, ES and TS; PThe replacement of SBM with ES enhanced oleic and linoleic acid and, particularly, CLA content. Intermediate values wereobserved for the TS group. CLA content (0.91, 0.62 and 0.56 g/100 g lipids, respectively, for ES, TS and SBM; Pincreased throughout the trial in all groups. ES also reduced the proportion of SFA with respect to SBM (65.2, 68.2 and70.9 g/100 g lipids, respectively, for ES, TS and SBM; Pin the same order; Pimproving the health-quality of milk. The various soybean products did not affect either metabolic profile (protein, urea,glucose, cholesterol, NEFA, triglycerides, liver parameters and mineral serum content) or rumen parameters (pH, ammoniaand VFAs). The replacement of SBM with ES and TS permitted an improvement in the nutritional properties of milkwithout negatively affecting animal performances.

Cattle establish firm dominance relationships through ritualised fights. This study aimed at investigating behaviours involved in dominance relationships and effect of factors such as weight, age and repeated fighting experience in fighting dynamics. Subject of the study was the Valdostana breed, whose cows assess dominance relationships in traditional competitions. Tournaments consist in rounds in which cows interact in pairs to assess dominance. Only winners participate in subsequent rounds. An amount of 120 fights involving 145 cows was retained, and winners (51 cows) were considered as focal individuals. An ethogram of agonistic interactions was established, including behaviours of different agonistic intensity as physical interactions (pushes, clashes), displays (threats, vocalisations), and non agonistic approaches. A transition diagram of behaviours showed a tendency to express firstly non agonistic approaches and lastly more aggressive clashes. A mixed linear model analysis on traits like competition intensity, duration, and type of behaviours expressed showed a significant effect of age difference on behaviours. However, the most important factor was the number of rounds performed: from the first to subsequent fights agonistic intensity and physical contacts increased, and displays reduced. This may be due either to the fact that more aggressive individuals were likely to be the winners, or that in higher rounds the opponents were more similar regarding fighting ability or aggressiveness and thus more intense fights occurred. The increased aggressiveness after repeated situations of competition suggests suggests that careful attention should be paid to welfare when animals are exposed to situations of high competition, like regrouping.

This study aimed to analyze the genetic variability of the Italian Heavy Draught Horse (IHDH) breed using a panel of 23 microsatellite markers. We also compared the population structure of the IHDH to other two unrelated breeds (Italian Haflinger, IH and Quarter Horse, QH). The IHDH showed a genetic variability comparable with other European heavy draught horse breeds and with the IH and QH breeds analyzed. Clustering analyses using a posterior Bayesian approach clearly differentiated the three breeds; it also showed a fragmentation of the IHDH in three subpopulations that need to be further investigated. These findings are an indicator of the present situation of the IHDH and will contribute to the conservation and implementation of the selection programme for this breed.

Performance testing is the main method of selection used for Marchigiana, Chianina and Romagnola breed since 1985. The young bulls are tested in a common central station for 6 months period, starting from the age of about 6 months. The bull selection is carried out accounting for genetic indexes of average daily gain (ADG) measured till weaning (PAGD), ADG realized during the test (TADG) and fleshiness (FLESH) measured at the end of test. Data of 2422 young bulls (735 Marchigiana, 863 Chianina and 824 Romagnola) were used to update estimates of heritability for performance test traits of aforementioned Italian beef cattle breeds. For traits measured till weaning (PADG), heritability differed among breeds from very low values for Marchigiana (0.02) to moderate for Romagnola (0.20), while for TADG, h2 estimates were more similar among breeds, ranging from 0.28 to 0.34. Similarly, h2 values obtained for FLESH trait were higher than values obtained till to weaning, but more variable among breed than TADG, i.e. from 0.19 to 0.55.

Performance test is the main selection tool for beef and dual purpose cattle, but it is used also in dairy cattle, despite the different aim of testing in this case. This study has aimed to obtain all available information about organisation and characteristic of performance test in Italy carried out in year 2006 by Italian Cattle Breeders Associations. The results of the survey here reported have the final aim of exploiting actual organization of selection processes carried out in Italy for beef, dual purpose and dairy cattle breeds through performance test. The survey has indicated different intensity of selection among Italian breeds, ranging from 2.45 to 3.13.

From 2001 to 2003 health data of 588 groups of fattening beefs (38723 animals) of 29 farms and 5 genetic types (Charolais, Female Charolais, French Crosses, Limousine and Polish Friesian) were collected and statistically analysed considering farm, breed, initial body weight, housing system, season, year, diet, average daily gain and density. The main causes of culling were problems of the locomotor and respiratory systems. As regards the genetic type and the initial body weight, the Polish Friesian breed and the lightest weights increased the probability of dying or getting injured respectively of 70 and 34%, while this probability decreased of 8.5% for every 100 g of daily growth higher than the mean value. As regards the other factors, bedding reduced the risk of culling by 33% in comparison with the slatted floor, while this risk increased of 10% for every m2 available/animal over the mean density.

This study has aimed to compare Repeatability (RP-TDm) and Random-Regression Test Day models (RR-TDm) in genetic evaluations of milk (M), fat (F) and protein (P) yields in Rendena breed. Variance estimations for Milk (M), Fat (F) and Protein (P) were obtained on a sample of 43,842 TD belonging to 2,692 animals controlled over 15 years (1990-2005). RP-TDm estimates of h2 were of 0.21 for M and 0.17 for both F and P, whereas RR-TDM provided a trend of h2 ranging from 0.15-0.34 for M, 0.15-0.31 for F and 0.10-0.24 for P. Both RP-TDm and RR-TDm results agreed with literature, even though RR-TDm provided a pattern of h2 along the lactation different from other studies, with the lowest h2 at the beginning and at the end of lactation. PSB, MAD and -2Log L parameters revealed lower power of RP-TDm as compare with the RR-TDm.

In this study, two different methods of scoring were implemented and compared in reason to assign a correct phenotypic value to cow fighting ability, and the variance components of the trait were estimated. Moreover, it was investigated the correlation of the estimated breeding values (EBVs) for fighting with the EBVs for the linear type traits muscularity. A scoring method mainly based on position (PS) and a method considering opponents’ performances (CPS) were compared and used in genetic analyses. A correlation of 0.75 between EBVs associated to PS and CPS was obtained, and ANOVAs considering alternatively PS and CPS indicated almost the same magnitude of the different sources of variation included (R2=0.50 and 0.52 for PS and CPS, respectively). Heritability estimates with the REML method were 0.099 (S.E.=0.005) for PS and 0.034 (S.E.=0.007) for CPS. Pearson and Spearman correlations between both indexes and ranks were no significant. Finally, an evaluation of the fitness of the two models revealed that PS is the better system for scoring, although CPS was designed to account also for the opponents.

The current studies define the role of three distinct cis-elements in the proximal promoter of the rat farnesyl diphosphate (FPP) synthase gene. The three cis-elements, a sterol regulatory element 3 (SRE-3) flanked by an ATTGG motif (inverted CCAAT box), and a CCAAT box, form a sterol regulatory unit that is necessary and sufficient for sterol-regulated expression of FPP synthase promoter-reporter genes. FPP synthase promoter-reporter genes, that contain promoters with either wild-type nucleotide sequences or mutations in one or more of the three cis-elements, were transiently transfected into CV-1 cells. The activity of the wild-type promoter-reporter gene increased when the cells were incubated in sterol-depleted media or when the cells were co-transfected with a plasmid encoding the mature form of SRE binding protein (SREBP-1a). The results with the mutant promoter-reporter genes demonstrated that all three cis-elements were necessary for normal expression/regulation of the reporter gene by either sterols or by co-expressed SREBP-1a. Gel mobility shift assays demonstrated that the synergistic binding of SREBP-1a to SRE-3 was dependent on the binding of recombinant nuclear factor Y (NF-Y) to the DNA, consistent with the in vivo regulation studies.—Jackson, S. M., J. Ericsson, R. Mantovani, and P. A. Edwards. Synergistic activation of transcription by nuclear factor Y and sterol regulatory element binding protein. J. Lipid Res. 1998. 39: 767–776.