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Submitted 2020-06-30 | Accepted 2020-07-25 | Available 2020-12-01 https://doi.org/10.15414/afz.2020.23.mi-fpap.67-73 Milk is considered not only a source of nutrient for the offspring but also a font of immunoregulatory compounds capable to predispose the naive intestinal immune system of the new-born to react to the external environment. In the present study we evaluated the composition of milk microbiome from cows classified according to milk total and differential somatic cell counts. A total of 34, 13 and 13 milk samples of healthy, at risk and subclinical or chronic cows, respectively, were collected during the same milking from a local dairy herd. Through Next Generation Sequencing (NGS) of bacterial 16S rRNA gene, the differences of taxa in terms of relative abundances (RA) and alpha and beta biodiversity were analysed. The RA of several genera were statistically significant in the three groups, such as Arcanobacterium (p=0.001), Rhodococccus (p

Simple Summary Along with the traditional traits, swine breeding programs for Italian dry-cured ham production have recently aimed to include novel phenotypes. The identification of the genomic regions underlying such new traits helps to untangle their genetic architecture and may provide useful information to be integrated in genetic selection. With this aim, we estimated genetic parameters and conducted a single step genome wide association studies (GWAS) on untrimmed and trimmed thigh weight considering two pig crossbred lines approved for Italian Protected Designation of Origin ham production. Quantitative trait loci (QTLs) were characterized based on the variance of 10-SNP sliding windows genomic estimated breeding values. In particular, we identified interesting QTL signals on several chromosomes, notably on chromosome 4, 6, 7 and 15. A high heritability and genetic correlation were observed for the two traits under investigation and although independent studies including other pig populations are required to disentangle the possible effects of specific linkage disequilibrium in our population, our findings suggest that such QTL could be investigated in future pig breeding programs to improve the reliability of genomic estimated breeding values for the dry-cured ham production. Abstract Protected Designation of Origin (PDO) dry-cured ham is the most important product in the Italian pig breeding industry, mainly oriented to produce heavy pig carcasses to obtain hams of the right weight and maturity. Recently, along with the traditional traits swine breeding programs have aimed to include novel carcass traits. The identification at the genome level of quantitative trait loci (QTLs) affecting such new traits helps to reveal their genetic determinism and may provide information to be integrated in prediction models in order to improve prediction accuracy as well as to identify candidate genes underlying such traits. This study aimed to estimate genetic parameters and perform a single step genome wide association studies (ssGWAS) on novel carcass traits such as untrimmed (UTW) and trimmed thigh weight (TTW) in two pig crossbred lines approved for the ham production of the Italian PDO. With this purpose, phenotypes were collected from ~1800 animals and 240 pigs were genotyped with Illumina PorcineSNP60 Beadchip. The single-step genomic BLUP procedure was used for the heritability estimation and to implement the ssGWAS. QTL were characterized based on the variance of 10-SNP sliding window genomic estimated breeding values. Moderate heritabilities were detected and QTL signals were identified on chromosome 1, 4, 6, 7, 11 and 15 for both traits. As expected, the genetic correlation among the two traits was very high (~0.99). The QTL regions encompassed a total of 249 unique candidate genes, some of which were already reported in association with growth, carcass or ham weight traits in pigs. Although independent studies are required to further verify our findings and disentangle the possible effects of specific linkage disequilibrium in our population, our results support the potential use of such new QTL information in future breeding programs to improve the reliability of genomic prediction.

Mastitis is an inflammatory disease of the mammary gland, caused by the invasion of microorganism on this site, associated with an altered immune response. Recent studies in this field hypothesize that the origin of these pathogens can also be from the gastrointestinal tract, through the entero-mammary pathway in relation to an increase in gut permeability. In this study, we wanted to investigate if inflammatory status of the mammary gland is related to an alteration of gut permeability. The microbiome of feces, blood and milk of lactating cows, recruited on the basis of the total somatic cell count and of the percentage of polymorphonuclear neutrophils and lymphocytes, was studied. Cows were divided into healthy (G), at risk of mastitis (Y) and with mastitis (R) classifications. The bacterial DNA was extracted and the V3 and V4 regions of 16S rRNA sequenced. Moreover, the quantification of total bacteria was performed with quantitative real-time PCR. A non-parametric Kruskal–Wallis test was applied at the phylum, family and genera levels and beta biodiversity was evaluated with the unweighted UniFrac distance metric. Significant differences between groups were found for the microbial composition of feces (Clostridiaceae, Turicibacteriaceae for family level and Clostridium, Dorea, SMB53 and Turicibacter for genus level), blood (Tenericutes for phylum level and Mycoplasma for genus level) and milk (OD1 and Proteobacteria for phylum level, Enterobacteriaceae and Moraxallaceae for family level and Olsenella and Rhodococcus for genus level). The beta biodiversity of feces and blood did not change between groups. Significant differences (p &lt, 0.05) were observed between the beta diversity in milk of G group and Y group and between Y group and R group. The number of taxa in common between feces, blood and milk were 8 at a phylum, 19 at a family and 15 at a genus level. From these results, the bacterial crossing from gut to milk in cows was not confirmed but remained hypothetical and deserves further investigation.

Stress in livestock reduces productivity and is a welfare concern. At a physiological level, stress is associated with the activation of inflammatory responses and increased levels of harmful reactive oxygen species. Biomarkers that are indicative of stress could facilitate the identification of more stress-resilient animals. We examined twenty-one metabolic, immune response, and liver function biomarkers that have been associated with stress in 416 Italian Simmental and 436 Italian Holstein cows which were genotyped for 150K SNPs. Single-SNP and haplotype-based genome-wide association studies were carried out to assess whether the variation in the levels in these biomarkers is under genetic control and to identify the genomic loci involved. Significant associations were found for the plasma levels of ceruloplasmin (Bos taurus chromosome 1—BTA1), paraoxonase (BTA4) and γ-glutamyl transferase (BTA17) in the individual breed analysis that coincided with the position of the genes coding for these proteins, suggesting that their expression is under cis-regulation. A meta-analysis of both breeds identified additional significant associations with paraoxonase on BTA 16 and 26. Finding genetic associations with variations in the levels of these biomarkers suggests that the selection for high or low levels of expression could be achieved rapidly. Whether the level of expression of the biomarkers correlates with the response to stressful situations has yet to be determined.

Simple Summary In recent years, the meat industry is looking with increased interest at the implementation of non-invasive new tools for predicting the final quality of dry-cured hams and monitoring ham aging. The selection of raw meat and the control of the salting procedure to predict the quality of dry-cured hams are of primary importance for meat processors. The identification of genetic markers associated with ham traits and related to different aptitudes of the thighs towards salting phases and weight losses is a primary goal for the Italian pig production chain. This paper addresses the need for investigating the associations between genomic markers and ham traits obtained through the application of non-invasive technologies for monitoring hams before and during the salting process. To our knowledge, this is the first study investigating the markers and genes associated with ham traits obtained through the use of Ham InspectorTM apparatus. Abstract Protected Designation of Origin (PDO) dry-cured hams production is greatly dependent on raw meat quality. This study was performed to identify genetic markers associated with the quality of dry-cured ham. Carcass traits of 229 heavy pigs belonging to three commercial genetic lines were registered (weight, EUROP classification). Phenotypic traits (Semimembranosus muscle ultimate pH, ham weight and lean meat content, adsorbed salt) of the corresponding thighs, undergone PDO ham process in three different plants, were measured, using a fast and non-invasive technology. Green ham weight and lean meat percentage influenced the estimated salt content and the weight loss during salting, even if the processing plant greatly affected the variability of the measured ham traits. The genomic data were obtained with the GeneSeek Genomic Profiler (GGP) 70k HD Porcine Array, using the slaughter day and the sex of the animals in the statistical analyses. The phenotypic traits were associated with the genotypes through GenAbel software. The results showed that 18 SNPs located on nine porcine chromosomes were found to be associated with nine phenotypic traits, mainly related to ham weight loss during salting. New associations were found between markers in the genes Neural Precursor Cell Expressed Developmentally Down-Regulated 9 (NEDD9, SSC7), T-Cell Lymphoma Invasion and Metastasis 2 (TIAM2, SSC1), and the ham quality traits. After validation, these SNPs may be useful to improve the quality of thighs for the production of PDO dry-cured hams.

Simple Summary Bovine milk contains proteins, minerals (e.g., calcium), vitamins (A, B, D, E and K), bioactive compounds and signaling molecules, as short non-coding RNA, either floating or contained in exosomes. Milk compositions and the milk-derived exosomes’ (EXO) cargo varies in relation to the health status of the mammary gland and the lactation stage. Stress, immune-depression and mastitis generate a differential microRNA (miRNA) expression, and consequently a modulation of the local immune cell response. This review aims to discuss the possible application of EXO cargo as a biomarker of animal resilience, and as a phenotype for genomic studies. Abstract Recent advances in ruminants’ milk-derived exosomes (EXO) have indicated a role of microRNAs (miRNAs) in cell-to-cell communication in dairy ruminants. The miRNAs EXO retain peculiar mechanisms of uptake from recipient cells, which enables the selective delivery of cargos, with a specific regulation of target genes. Although many studies have been published on the miRNAs contained in milk, less information is available on the role of miRNAs EXO, which are considered stable over time and resistant to digestion and milk processing. Several miRNAs EXO have been implicated in the cellular signaling pathway, as in the regulation of immune response. Moreover, they exert epigenetic control, as extenuating the expression of DNA methyltransferase 1. However, the study of miRNAs EXO is still challenging due to the difficulty of isolating EXO. In fact, there are not agreed protocols, and different methods, often time-consuming, are used, making it difficult to routinely process a large number of samples. The regulation of cell functions in mammary glands by miRNAs EXO, and their applications as genomic markers in livestock, is presented.

Several studies have underlined the interplay among host-microbiome and pathophysiological conditions of animals. Research has also focused specifically on whether and how changes in the gut microbiome have provoked the occurrence of pathological phenomena affecting cartilage and joints in humans and in laboratory animals. Here, we tried to evaluate the relationship between the gut microbiome and the hip and elbow arthritis in owned dogs. The study included 14 dogs suffering from chronic arthritis (AD) and 13 healthy dogs (HD). After the first visit and during the period of the study, the dogs, under the supervision of the owner, were fed a semi-moist complete diet supplemented with omega 3 fatty acids. Feces and blood samples were collected in the clinic at the first visit (T0) and after days (T45). The plasma C-reactive protein (CRP) was higher, and the serum vitamin B12 and folate concentrations were lower (p < 0.05) in the AD group in comparison to the HD group. Data of the fecal microbiome showed that the relative abundances of the genus Megamonas were higher in AD (p < 0.001), while the relative abundance of the families Paraprevotellaceae, Porphyromonadaceae, and Mogibacteriaceae was significantly lower in comparison to HD. The results of the study identified several bacterial groups that differed significantly in the fecal microbiome between healthy and diseased dogs. If the observed differences in fecal bacterial composition predispose dogs to hip and elbow arthritis or if these differences reflect a correlation with these conditions deserves further investigation.

Dairy cattle health, wellbeing and productivity are deeply affected by stress. Its influence on metabolism and immune response is well known, but the underlying epigenetic mechanisms require further investigation. In this study, we compared DNA methylation and gene expression signatures between two dairy cattle populations falling in the high- and low-variant tails of the distribution of milk cortisol concentration (MC), a neuroendocrine marker of stress in dairy cows. Reduced Representation Bisulfite Sequencing was used to obtain a methylation map from blood samples of these animals. The high and low groups exhibited similar amounts of methylated CpGs, while we found differences among non-CpG sites. Significant methylation changes were detected in 248 genes. We also identified significant fold differences in the expression of 324 genes. KEGG and Gene Ontology (GO) analysis showed that genes of both groups act together in several pathways, such as nervous system activity, immune regulatory functions and glucocorticoid metabolism. These preliminary results suggest that, in livestock, cortisol secretion could act as a trigger for epigenetic regulation and that peripheral changes in methylation can provide an insight into central nervous system functions.

A dietary intervention study was assessed to determine if different sources of starch in homemade diets could significantly modify fecal microbiome of dogs. Twenty-seven adult dogs were enrolled and fed a diet based on a mixture of rice and pasta with fresh raw meat (CD). After 90 d, 8 dogs continued to receive CD diet, 10 dogs received a diet made of a raw meat and a complementary food with rice as the main source of starch (B1), and 9 dogs were fed a diet with the same raw meat and a complementary food with potato as the main source of starch (B2). Samples of feces were collected from each dog in the mornings at the beginning of the study and after 15 d and analyzed for pH, ammonia N (N–NH3) and total N, short chain fatty acids (SCFA) and lactic acid. Relative abundance of fecal microbiota was assessed by sequencing and annotating the V3–V4 regions of the 16S rRNA. Total starch intake was similar between diets but differed in the in vitro rate digestion and in the resistant starch, which was higher in B2 than in B1 and CD diets. Dogs fed B2 diet showed lower (P

Genomic selection (GS) reports on milk fatty acid (FA) profiles have been published quite recently and are still few despite this trait represents the most important aspect of milk nutritional and sensory quality. Reasons for this can be found in the high costs of phenotype recording but also in issues related to its nature of complex trait constituted by multiple genetically correlated variables with low heritabilities. One possible strategy to deal with such constraint is represented by the use of dimension reduction methods. We analysed 40 individual FAs from Italian Brown Swiss, Holstein and Simmental milk through multivariate factor analysis (MFA) to study the genetics of milk FA-related latent variables (factors) and assess their potential use in breeding. A total of nine factors were obtained, and their genetic parameters were inferred under a Bayesian framework using two statistical approaches: the classical pedigree best linear unbiased prediction (ABLUP) and the single-step genomic BLUP (ssGBLUP). The resulting factorial solutions were able to represent groups of FAs with common origin and function and can be considered concise pathway-level phenotypes. The heritability (h2 ) values showed relevant variations across different factors in each breed (0.03 ≤ h2 ≤ 0.38). The accuracies of breeding values predicted were low to high, ranging from 0.13 to 0.72 and from 0.18 to 0.74 considering the pedigree and the genomic model, respectively. The gain in accuracy in genetic prediction due to the addition of genomic information was ~30% and ~5% in validation and training groups respectively, confirming the contribution of genomic information in yielding more accurate predictions compared to the traditional ABLUP methodology. Our results suggest that MFA in combination with GS can be a valuable tool in dairy cattle breeding and deserves to be further investigated for use in future breeding programs to improve cow's milk FA-related traits.

The characterization of the microbial population of many niches of the organism, as the gastrointestinal tract, is now possible thanks to the use of high-throughput DNA sequencing technique. Several studies in the companion animals field already investigated faecal microbiome in healthy or affected subjects, although the methodologies used in the different laboratories and the limited number of animals recruited in each experiment does not allow a straight comparison among published results. In the present study, we report data collected from several in house researches carried out in healthy dogs, with the aim to seek for a variability of microbial taxa in the faeces, caused by factors such as diet and sex. The database contains 340 samples from 132 dogs, collected serially during dietary intervention studies. The procedure of samples collection, storage, DNA extraction and sequencing, bioinformatic and statistical analysis followed a standardized pipeline. Microbial profiles of faecal samples have been analyzed applying dimensional reduction discriminant analysis followed by random forest analysis to the relative abundances of genera in the feces as variables. The results supported the responsiveness of microbiota at a genera taxonomic level to dietary factor and allowed to cluster dogs according this factor with high accuracy. Also sex factor clustered dogs, with castrated males and spayed females forming a separated group in comparison to intact dogs, strengthening the hypothesis of a bidirectional interaction between microbiota and endocrine status of the host. The findings of the present analysis are promising for a better comprehension of the mechanisms that regulate the connection of the microorganisms living the gastrointestinal tract with the diet and the host. This preliminary study deserves further investigation for the identification of the factors affecting faecal microbiome in dogs.

Milk fatty acid (FA) profile is a clear example of complex and multiple correlated traits whose genetic basis is difficult to assess. Although genome-wide association (GWA) studies have been successful in the identification of significant genetic variants for complex traits, when correlated phenotypes are analysed separately, the outcomes are difficult to compare and interpret in a metabolic context. Here, we performed a multivariate factor analysis (MFA) on Italian Simmental and Italian Holstein milk fat profiles to extract latent unobserved factors able to explain correlation structure and common metabolic function among different FAs. Individual factor scores obtained by MFA were used to perform a single-SNP based GWA. In both breeds, MFA was able to extract ten latent factors with specific biological meaning, notably: de novo synthesis, desaturation activity and biohydrogenation. The GWA result confirmed the increased power of joint association analysis on multiple correlated traits and allowed us to identify major candidate genes with well-documented function consistent with the metabolic classification of factors obtained, such as DGAT1, FASN and SCD.

Several studies on the interaction between gut microbiota and diets, including prebiotics, have been reported in dogs, but no data are available about the effects of dietary administration of grape proanthocyanidins. In the study, 24 healthy adult dogs of different breeds were recruited and divided in 3 groups of 8 subjects each. A group was fed with a control diet (D0), whilst the others were supplemented with 1 (D1) or 3 (D3) mg/kg live weight of grape proanthocyanidins. Samples of feces were collected at the beginning and after 14 and 28 days for microbiota, short chain fatty acid, and lactic acid analysis. Serotonin and cortisol were measured in saliva, collected at the beginning of the study and after 28 days. A significantly higher abundance (p &lt, 0.01) of Enterococcus and Adlercreutzia were observed in D0, whilst Escherichia and Eubacterium were higher in D1. Fusobacterium and Phascolarctobacterium were higher (p &lt, 0.01) in D3. Salivary serotonin increased (p &lt, 0.01) at T28 for D1 and D3 groups but cortisol did not vary. Proanthocyanidins administration influenced the fecal microbiota and neuroendocrine response of dogs, but a high variability of taxa was observed, suggesting a uniqueness and stability of fecal microbiota related to the individual.

The characterization of the microbial population in different compartments of the organism, such as the gastrointestinal tract, is now possible thanks to the use of high-throughput DNA sequencing technique. Several studies in the companion animals field have already investigated the fecal microbiome in healthy or sick subjects; however, the methodologies used in the different laboratories and the limited number of animals recruited in each experiment do not allow a straight comparison among published results. Previously, our research focused on the characterization of the microbial taxa variability in 340 fecal samples from 132 healthy dogs, collected serially from several in-house experiments. The results supported the responsiveness of microbiota to dietary and sex factors and allowed us to cluster dogs with high accuracy. For the present study, intestinal and blood microbiota of healthy dogs from different breeds, genders, ages and food habits were collected, with three principal aims: firstly, to confirm the results of our previous study regarding the fecal microbiome affected by the different type of diet; secondly, to investigate the existence of a blood microbial population, even in heathy subjects; and thirdly, to seek for a possible connection between the fecal and the blood microbiota. Limited researches have been published on blood microbiota in humans, and this is the first evidence of the presence of a bacterial population in the blood of dogs. Moreover, gut and blood microbiota can discriminate the animals by factors such as diet, suggesting some relationship between them. These preliminary results make us believe in the use of the blood microbiome for diagnostic purposes, such as researching and preventing gut inflammatory diseases.

The study aimed at investigating the rumen microbiome composition and functional activity in mid lactating cows of Italian Holstein (IH) and Italian Simmental (IS) breeds. Eight IH and eight IS pluriparous cows with days in milking (DIM) ranging from 90 to 180 were selected and rumen contents were sampled with oesophageal tube. Rumen metagenome was analysed using a whole shotgun sequencing. Data were analysed for taxonomic classification and microbial genes. The relative abundance of Archaea, the Archaea to Bacteria ratio and the Archaea to Eukarya ratio were higher (p

Ruminant production has been an essential part of human activities worldwide since ancient times. The expected increase in world population and per capita income, with an increase in the amount and prevalence of animal products in human diet, urbanisation, with a concentration of population in urban areas and an increase in losses in the supply chain, and the growing concern over the environmental impact of animal farming require a long-term global strategy for a more intensive and sustainable ruminant production. Therefore, solutions to increase the supply of high-quality products of ruminant origin, without harming human health, animal welfare, and environment, should consider the following interconnected issues discussed in this review: (a) effects of meat, milk and dairy products consumption on human health, focussing on the imbalance caused by their insufficient consumption, and the alleged increased incidence of certain diseases due to their consumption; (b) importance of the sustainable intensification of ruminant production systems (e.g. better feed conversion and higher production output per unit of input introduced into the farming system); (c) environmental impact of ruminant production; (d) improvement of animal performance by improving animal welfare; (e) adaptation of ruminants to climate change; (f) sustainable ruminant feeding (e.g. precision feeding techniques, optimisation of grazing systems, and use of unconventional feeds); (g) challenges posed by production intensification to animal breeding and conservation of animal biodiversity; and (h) strategies to increase ruminant production in developing countries, thus achieving food security in vast areas of the planet affected by fast growth of human population.

The aim of the study was to investigate element concentrations in the hair of growing and adult dogs. Overall, 39 dogs were recruited in a kennel, split in 4 groups: G1, 10 puppies of small size breeds; G2, 6 puppies of medium size breed; G3, 11 adult dogs of medium size breeds; G4, 12 adult dogs of toy breeds. The dogs of each group were fed 4 different complete diets, according to the requirements. Hairs were sampled at the beginning of the study (D0) and after 60 days (D60). Elements were analysed in triplicate using ICP-OES. In the hair of the adult dogs of the G3 group the concentration of Al (p

The aim of the study was to investigate if the inclusion of chickpeas or peas in the diet can modify faecal microbiome in dogs. Eight healthy adult Border collie, fed a commercial extruded diet as reference diet (RD), were divided in two groups of four individuals. At the beginning of the trial, one group received a diet based mainly of raw meat, rice and chickpeas (CP) and in the other group this pulse was substituted with peas (PE). After 14 days, the dogs with CP diet shifted to the PE and those with PE shifted to the CP diet, for another 14 days. Faeces were collected at the beginning (T0), after 14 days (T14) and at the end of the study (T28). Faeces were analysed for 16S rRNA, short chain fatty acids (SCFA), lactate, pH and faecal score was also evaluated. The SCFA and lactate in the faeces were not affected by the inclusion of pulses, with the only exception of isovalerate, which was higher in CP and PE diets in comparison with RD diet (p < .05). The abundances of Erysipelotrichaceae incertae sedis, Eubacterium, Anaerobacter and Sarcina significantly differed in CP and PE in comparison with RD. Moreover, the genera Prevotella, Lactobacillus, Alloprevotella, Suttarella varied significantly between CP and PE diets. The observed modifications of faecal microbioma were related not only to the change from RD to CP or PE, but also to the type of pulse, chickpeas or peas. However, long-term studies are required to investigate the implications that pulses can have for gut health.Highlights Faecal quality and end products of fermentation were similar between diets. Faecal microbiota is affected by the type of pulse and raw meat inclusion. Thermally treated Chickpeas or Peas in the diet of dogs were safe, but further study are required.

This study examined microRNA (miRNAs) expression and regulatory patterns from milk exosomes of cows experienced group relocation, a common husbandry practice during the lactation period and used as a spontaneous model of stress. Total RNA from milk exosome samples was collected from 3 cows that showed an increased milk cortisol (HRC) after relocation (T2 vs T1) and 3 cows that did not show cortisol increase (LRC). A total of 69 known miRNAs were identified. Thirteen miRNAs were consistently down-regulated at T2 in comparison to T1, such as miR-2904-1, miR-142, miR-2284x and miR-30b-3p. Only two miRNAs, miR-2284z and miR-146a were significantly different between LRC and HRC group. Functional enrichment analyses highlighted that glucocorticoid receptor signaling and neurotrophic factor mediated TRK receptor signaling were among the biological pathways affected by differentially expressed miRNA target genes. Mir-135a-5p and miR-320a were involved in both biological pathways. miR-142-5p, miR-142-3p, miR-30b-5p and miR-320a shared the same target genes belonging to the RAS superfamily of small GTP-binding proteins (RAC1, RAP1A and RASA1), involved in neurotrophin-mediated cell survival. MiR-142, miR-135 and miR-320a in milk exosomes were the most responsive to group relocation of cows.

Bovine milk is important for human nutrition, but its fat content is often criticized as a risk factor in cardiovascular disease. Selective breeding programs could be used to alter the fatty acid (FA) composition of bovine milk to improve the healthiness of dairy products for human consumption. Here, we performed a genome-wide association study (GWAS) on bovine milk to identify genomic regions or specific genes associated with FA profile and to investigate genetic differences between the Italian Simmental (IS) and Italian Holstein (IH) breeds. To achieve this, we first characterized milk samples from 416 IS cows and 436 IH cows for their fat profile by gas chromatography. Subjects were genotyped with single nucleotide polymorphism array and a single-marker regression model for GWAS was performed. Our findings confirm previously reported quantitative trait loci strongly associated with bovine milk fat composition. More specifically, our GWAS results revealed significant signals on chromosomes Bos taurus autosome 19 and 26 for milk FA. Further analysis using a gene-centric approach and pathway meta-analysis identified not only some well-known genes underlying quantitative trait loci for milk FA components, such as FASN, SCD, and DGAT1, but also other significant candidate genes, including some with functional roles in pathways related to "Lipid metabolism." Highlighted genes related to FA profile include ECI2, PCYT2, DCXR, G6PC3, PYCR1, and ALG12 in IS, and CYP17A1, ACO2, PI4K2A, GOT1, GPT, NT5C2, PDE6G, POLR3H, and COX15 in IH. Overall, the breed-specific association outcomes reflect differences in the genetic backgrounds of the IS and IH breeds and their selective breeding histories.

The aim of the research was to investigate the variation of gene expression in the blood of lactating cows in relation to the genetic merit for productive traits. For the study, 24 Italian Holstein (IH) cows from a single farm and 24 Italian Simmental (IS) cows from a second farm were selected. Cows were in mid-lactation, and farms had similar management and feeding conditions. For each breed, cows were separated into 3 classes of estimated breeding value (EBV) for milk protein yield (EBVp), namely, 8 cows with low EBVp (low group, LG), 8 cows with high EBV (high group, HG), and the 8 cows closest to the median (medium group, MG). Gene expression was measured on blood with a whole-transcriptome bovine microarray, and data of LG and HG were expressed relative to MG. The number of differentially expressed genes between the low and high EBVp groups were 443 for IS and 281 for IH cows. The IS cows had a greater number of genes (398 vs. 241 in IH cows) with opposite expression in the high and low EBV groups, compared with the MG. In IS cows, the largest number of pathways affected were within the KEGG subcategories of "energy metabolism," "lipid metabolism," and "metabolism of cofactors and vitamins." Furthermore, the "glutathione metabolism" pathway was markedly affected, with GSTM1 (from glutathione-S-transferase family), GSS (glutathione synthetase), and G6PD (glucose-6-phosphate dehydrogenase) upregulated in the IS HG but downregulated in the IS LG group. The "intestinal immune network for IgA production" pathway had 2 bovine leukocytes antigen ( BoLA-DQ ) genes, and the B-cell activating factor of the TNF superfamily ( TNFSF13B ) upregulated in the IS LG group and downregulated in the IS HG group. The IH cows had the highest number of affected pathways in the "metabolism" and "organismal systems" categories, the latter with 5 out of 10 subcategories relative to the immune system. Within the "T cell receptor signaling pathway," the IH HG cows had upregulation of the CD1d , CD4 , and CD3 antigens and downregulation of the NFkB subunit ReLa gene. This study revealed that specific metabolic and immunological pathways are directly related to genetic merit, and could be considered a signature of quantitative selection. Starting from these preliminary observations, the comparison of differentially expressed genes among animals with different EBVp seems a promising approach to unravel molecular response at a blood level associated with productive traits.

The prevalence of obesity is increasing all over the world. Although it has been shown that natural substances influence fat metabolism, little is known about the effect on cellular and molecular mechanisms in human. In this in vitro study, the activity of Rosmarinus officinalis (RO) standardized extract in modulating human primary visceral preadipocytes differentiation, lipolysis, and apoptosis was investigated. Moreover, gene expression of key adipogenesis modulators and microRNAs-seq were evaluated. Preadipocytes treated with RO extract significantly reduced triglyceride incorporation during maturation in a dose-dependent manner without affecting cell viability. In addition, RO extract stimulated lipolytic activity in differentiating preadipocytes and mature adipocytes in treated cells compared to controls. Differentiating preadipocytes incubated in the presence of RO extract showed a decreased expression of cell cycle genes such as cyclin D1, cyclin-dependent kinase 4, cyclin-dependent kinase inhibitor 1A (p21, Cip1) and an increased expression of GATA binding protein 3, wingless-type MMTV integration site family, member 3A mRNA levels. Recent studies have demonstrated that some phytochemicals alter the expression of specific genes and microRNAs that play a fundamental role in the pathogenesis of obesity and related diseases. Interestingly, genes modulated in RO-treated cells were found to be validated miRNAs targets, such as let-7f-1, miR-17, and miR-143. The results indicated that RO extract modulates human adipocyte differentiation and significantly interferes with adipogenesis and lipid metabolism, supporting its interest as dietary supplement.

Background Dietary intervention studies are required to deeper understand the variability of gut microbial ecosystem in healthy dogs under different feeding conditions and to improve diet formulations. The aim of the study was to investigate in dogs the influence of a raw based diet supplemented with vegetable foods on faecal microbiome in comparison with extruded food. Methods Eight healthy adult Boxer dogs were recruited and randomly divided in two experimental blocks of 4 individuals. Dogs were regularly fed a commercial extruded diet (RD) and starting from the beginning of the trial, one group received the raw based diet (MD) and the other group continued to be fed with the RD diet (CD) for a fortnight. After 14 days, the two groups were inverted, the CD group shifted to the MD and the MD shifted to the CD, for the next 14 days. Faeces were collected at the beginning of the study (T0), after 14 days (T14) before the change of diet and at the end of experimental period (T28) for DNA extraction and analysis of metagenome by sequencing 16SrRNA V3 and V4 regions, short chain fatty acids (SCFA), lactate and faecal score. Results A decreased proportion of Lactobacillus, Paralactobacillus (P

The aim of this study was to investigate the adipocyte size and fate in subcutaneous fat (scAT) of cows diverging for genetic merit at mid lactation stage, when anabolic activity increases and animals are in a state of positive energy balance. Twenty mid lactation cows (180±20days in milk) grouped according to the Estimated Breeding Values (EBV) for milk yield in plus (EBVp) and minus (EBVm) variants were selected. Average of adipocytes area, proliferation and apoptotic labelling index as well as DLK-1 expression, a marker of pre-adipocytes, were immunohistochemically evaluated in scAT biopsies. In EBVp cows, the BCS was lower (P

The study was conducted to investigate the effects of dietary administrations of four nutraceuticals in dogs. Seventy four dogs were enrolled in the trials, 24 healthy dogs were fed with a control diet (CT) and the experimental groups received for 60days the same diet supplemented with nutraceuticals, namely Echinacea angustifolia (EA, 0.10mg/kg live weight as echinacoside; 14 dogs), Vaccinium myrtillus (VM, 0.20mg/kg live weight as anthocyanidin, 13 dogs), Curcuma longa (CL, 6.60mg/kg live weight as curcumin, 18 dogs with arthrosis), and Sylibum marianum (SM, 1.5mg/kg live weight as sylibin, 8 dogs with hepatopathy). Dogs were weighted at the beginning of study and blood samples were collected at the beginning (T0) and at the end (T60) of the study. VM significantly down regulated TNF, CXCL8, NFKB1 and PTGS2 and decreased plasma ceruloplasmin (CuCp). The activity of EA was evidenced by the significant decrease of TNF and NFKB1 expression and CuCp levels and by the increase of plasma Zn. Administration of CL caused a significant decrease of CuCp and increase of Zn and a down regulation of TNF, CXCL8, NFKB1 and PTGS2, corroborating the anti-inflammatory action of curcuminoids. After 60days of treatment with SM, plasma ALT/GPT activity was reduced and paraoxonase was increased, supporting the antioxidant activity of silymarin, also confirmed by the significant up regulation of SOD2. Results indicated that nutraceutical administrations in dogs can be an interesting approach to modulate immune response in order to improve health condition of animals.

Hydroxytyrosol has various pharmacological properties, including anti-oxidative stress and anti-inflammatory activities, preventing hyperglycemia, insulin resistance, and the metabolic syndrome. The present study is focused on the anti-adipogenic and lipolytic activity of hydroxytyrosol on primary human visceral adipocytes. Pre-adipocytes were analyzed after 10 (P10) and 20 (P20) days of treatment during differentiation and after 7 (A7) days of treatment when they reached mature shape. The treatment with hydroxytyrosol extract significantly ( P

Summary Using an array consisting of 10 665 70-mer oligonucleotide probes, the longissimus dorsi muscle tissue expression during growth in nine pigs belonging to Casertana (CT), an autochthonous breed characterized by slow growth and a massive accumulation of backfat, was compared with that of two cosmopolitan breeds, Large White (LW) and a crossbreed (CB; Duroc × Landrace × Large White). The results were validated by real-time PCR. All animals were of the same age and were raised under the same environmental conditions. Muscle tissues were collected at 3, 6, 9 and 11 months of age, and a total of 173 genes showed significant differential expression between CT and the cosmopolitan genetic types at 3 months of age. Time series cluster analysis indicated that the CT breed had a different pattern of gene expression compared with that of the LW and the CB. Four of the eight clusters highlighted the gene differences between CT and the other two breeds, which were further supported by statistical analyses: clusters 4 and 5 contained a total of 71 genes that were underexpressed at 3 months of age, and cluster 3 and cluster 7 included 28 and 42 genes respectively that were overexpressed at 3 months of age. As expected, differentially expressed genes belonged to the category of genes coding for contractile fibres and transcription factors involved in muscle development and differentiation. These findings highlight muscle expression genes during pig growth and are useful to understand the genetic meaning of the different developmental rates.

This review highlights fundamental mechanisms of the stress response and important findings as to how the immune system is affected and affects, in turn, such a response. The crucial link between stress response and energy metabolism is dealt with as well. The effector mechanisms in the stress response are remarkably similar for both infectious and non–infectious stimuli, albeit differently modulated. “Psychosensitive stimuli/behavioural response” and “Antigenic stimuli/immune response” are indeed two subsystems of a unitary, integrated complex aimed at providing optimal conditions for the host’s survival and adaptation. The interaction between the immune system and the stress/inflammation complex has led to the development of a diversified network of cytokines and chemokines in vertebrate animals. The cytokine response can be mounted in different forms and extent by the host after exposure to both infectious and non-infectious stimuli. In this conceptual framework, microbial infections are just one category of stressing agents, which modulate the cytokine response for a better performance of the innate and adaptive immune responses. The response to infectious and non–infectious stress leads to a metabolic shift that enhances energy, amino acids and micronutrients consumption. The influence of each nutrient on different aspects of immune function is not easy to define, but it is becoming clear that many nutrients have defined roles in the immune response and, accordingly, their requirements are changed to support optimal immune function. Therefore, impairment of immune functions may arise from intakes of nutrients below or above these modified ranges of requirements.

Neutrophils play an essential role in host defense and inflammation. Plants have long been used to improve the immune function, but for most of them specific investigations on animal health are lacking. In the present study, water and hydroethanolic extracts from 11 plant wastes have been screened on immune responses of ovine neutrophils. Eight sheep clinically healthy, not lactating, non-pregnant were selected and used for the experiment. Freshly isolated neutrophils were incubated with the extracts of the residues at increasing doses, and then they were tested for adhesion and superoxide production induced with PMA. The residues of Larix decidua, Thymus vulgaris, Salix alba, Sinupret, Helianthus annuus, Mangifera indica modulated the neutrophil immune functions, moreover, Larix decidua, Thymus vulgaris and Salix alba presented the highest anti-inflammatory activity.

The discovery of new protein-coding DNA variants related to carcass traits is very important for the Italian pig industry, which requires heavy pigs with higher thickness of subcutaneous fat for Protected Designation of Origin (PDO) productions. Exome capture techniques offer the opportunity to focus on the regions of DNA potentially related to the gene and protein expression. In this research a human commercial target enrichment kit was used to evaluate its performances for pig exome capture and for the identification of DNA variants suitable for comparative analysis. Two pools of 30 pigs each, crosses of Italian Duroc X Large White (DU) and Commercial hybrid X Large White (HY), were used and NGS libraries were prepared with the SureSelectXT Target Enrichment System for Illumina Paired-End Sequencing Library (Agilent). A total of 140.2 M and 162.5 M of raw reads were generated for DU and HY, respectively. Average coverage of all the exonic regions for Sus scrofa (ENSEMBL Sus_scrofa.Sscrofa10.2.73.gtf) was 89.33X for DU and 97.56X for HY; and 35% of aligned bases uniquely mapped to off-target regions. Comparison of sequencing data with the Sscrofa10.2 reference genome, after applying hard filtering criteria, revealed a total of 232,530 single nucleotide variants (SNVs) of which 20.6% mapped in exonic regions and 49.5% within intronic regions. The comparison of allele frequencies of 213 randomly selected SNVs from exome sequencing and the same SNVs analyzed with a Sequenom MassARRAY® system confirms that this "human-on-pig" approach offers new potentiality for the identification of DNA variants in protein-coding genes.

Genome-wide association studies (GWAS) have been widely applied to disentangle the genetic basis of complex traits. In cattle breeds, classical GWAS approaches with medium-density marker panels are far from conclusive, especially for complex traits. This is due to the intrinsic limitations of GWAS and the assumptions that are made to step from the association signals to the functional variations. Here, we applied a gene-based strategy to prioritize genotype-phenotype associations found for milk production and quality traits with classical approaches in three Italian dairy cattle breeds with different sample sizes (Italian Brown n = 745; Italian Holstein n = 2058; Italian Simmental n = 477). Although classical regression on single markers revealed only a single genome-wide significant genotype-phenotype association, for Italian Holstein, the gene-based approach identified specific genes in each breed that are associated with milk physiology and mammary gland development. As no standard method has yet been established to step from variation to functional units (i.e., genes), the strategy proposed here may contribute to revealing new genes that play significant roles in complex traits, such as those investigated here, amplifying low association signals using a gene-centric approach.

Background Whether the measurement of cortisol in dairy cows can be used as a biomarker of adverse environmental or pathophysiological conditions is still under of scientific debate. In these situations, several systems mainly the hypothalamic-pituitary-adrenal axis, the autonomic nervous system, and the immune system are recruited to reestablish homeostasis. A first aim of the present study was to compare milk and blood cortisol concentrations and to consider its variability in milk in relation to farm, milk yield and days in milk. A second study investigates the effects of breed, class of somatic cell count (SCC) and farm on milk cortisol levels in a larger number of cows and farms, with the aim to validate the results obtained in the pilot study. Methods For study 1, 135 cows were sampled from 2 Italian Simmental and 2 Italian Holstein commercial farms, whilst in the second study, 542 cows were sampled from 6 commercial farms of Italian Simmental and 499 cows from 4 commercial farms of Italian Holstein. Results In study 1, the values of cortisol content in milk were significantly higher in Holstein than Simmental cows. Significant differences between farms were observed for milk and plasma cortisol concentrations. Cortisol content in milk was not correlated to plasma content in study 1 and the mean milk to plasma cortisol ratio was about 1:30. In study 2, for Holstein cows, significantly higher values of milk cortisol in comparison to Simmental cows was reported. A significant effect of class of SCC was observed, cows belonging to class 3 (SCC higher than 400.000/ml) showed the highest mean values of milk cortisol. The farm effect was significant also in the study 2, confirming the results obtained in the first study. Conclusions Milk can be considered a preferential site of sampling in dairy cows to point out short term stimulation of the hypothalamic-pituitary-adrenal axis. Further studies are needed to investigate the physiological basis of the relationship between milk cortisol content and breed, milk yield and SCC to ascertain the relevance of milk cortisol to monitor the healthy status of mammary gland.

Currently, there are no reports of neurotrophins in adipose tissue of cows. The distribution of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and their high-affinity tyrosine kinase receptors TrkA and TrkB, was investigated by immunohistochemical method in the subcutaneous adipose tissue of cow at mid-lactation. Results revealed the localization of NGF and BDNF along the plasma membrane and cytoplasm of adipocytes. Neurotrophin receptors TrkA and TrkB showed moderate and strong positive staining in adipocytes, respectively. The expression of NGF, BDNF, TRKB--but not of TRKA--was also confirmed at transcriptional level by RT-PCR analyses. Considering the involvement of BDNF on fat metabolism and of NGF on activation of the sympathetic response in human and rodents, these neurotrophins could be related to lipogenesis and lipolysis occurring during lactation in cows. The local production of these neurotrophins supports their potential paracrine function for the regulation of adipocyte activity and deserve further investigations.

Rhodiola rosea (Rro) has been reported to have various pharmacological properties, including anti-fatigue, anti-stress and anti-inflammatory activity. It is also known to improve glucose and lipid metabolism, but the effects of Rhodiola rosea on adipocyte differentiation and metabolism are not still elucidated. In this study the anti-adipogenic and lipolytic activity of two extracts of Rhodiola rosea, containing 3% salidroside (RS) or 1% salidroside and 3% rosavines (RR) on primary human visceral adipocytes was investigated. Pre-adipocytes were analyzed after 10 and 20 days of treatment during differentiation and after 7 days of treatment when they reached mature shape. The RS extract significantly induced higher apoptosis and lipolysis in comparison to control cells and to RR extract. In contrast, RR extract significantly reduced triglyceride incorporation during maturation. Differentiation of pre-adipocytes in the presence of RS and RR extracts showed a significant decrease in expression of genes involved in adipocyte function such as SLC2A4 and the adipogenic factor FGF2 and significant increase in expression of genes involved in inhibition of adipogenesis, such as GATA3, WNT3A, WNT10B. Furthermore RR extract, in contrast to RS, significantly down-regulates PPARG, the master regulator of adipogenesis and FABP4. These data support the lipolytic and anti-adipogenetic activity of two different commercial extracts of Rhodiola rosea in primary human visceral pre-adipocytes during differentiation.

A total of 113 DNA polymorphisms in 32 candidate genes were pre-screened to assess their variability in 32 Italian Simmental bulls. A total of 64 variants successfully genotyped and having a minor allele frequency (MAF) higher than 0.03 were then analyzed in 477 Italian Simmental bulls. Among these 37 had a MAF ≥0.05 and were investigated for association with milk production and functional traits. Daughter yield deviations (DYD) were available for milk, protein and fat yield (MY, PY and FY, respectively), somatic cell count (SCC), udder score (US), feet and legs score (FLS), while for protein percent (PP), calving ease direct effect (CED), fertility (FE) and milkability (MA) associations were computed using (EBVs). Twelve SNPs were significantly associated with at least one trait ( P ≤0.05). The strongest association ( P =0.0009) was found between rs385640152 in the growth hormone receptor gene ( GHR ) and PP. Also marker rs42714483 in the thyroid hormone responsive gene ( THRSP ) was associated with PP ( P =0.004). Pleiotropic effects were detected only for β-lactoglobulin ( LGB or PAEP progestagen-associated endometrial protein) polymorphisms on. SNP rs29004488 in the leptin gene ( LEP ) and rs41974998 in milk fat globule-epidermal growth factor 8 protein gene ( MFGE8 ) were associated with MY ( P =0.043 and P =0.033, respectively). Suggestive associations with functional traits were also detected: rs43347906 in the leptin receptor gene ( LEPR ) with US ( P =0.029), rs29004485 in LEP with FLS ( P =0.049), rs41703837 in the v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog ( SRC ) with CED ( P =0.021) and rs109019599 in the oxidized low density lipoprotein receptor 1 gene ( OLR1 ) with MA ( P =0.024). Our results both provide loci newly associated with functional traits and confirm associations already detected in Italian Holstein and Italian Brown breeds.

During the peripartum period, high-yielding dairy cows experience metabolic stress, which alters their homeostasis and exposes the cows to illness. The aim of this study was to quantify the expression levels of genes involved in antioxidant defences during the transition period in the blood of dairy cows and to evaluate the regulative activity on these genes of natural antioxidants in the diet. Three groups of 7 heifers each, at the 7th month of pregnancy, were used. Starting from 3 weeks before the expected calving date (–22 days), the three groups were allotted to the following experimental treatments: control (CTR, basal diet); lycopene (LYC, basal diet + lycopene 540 mg/day) and grape polyphenols (POL, basal diet + grape polyphenols 10 g/day). Blood was sampled at 22 and 8 days before and 8, 15 and 22 after calving and analysed for the expression level of glutathione peroxidase (GPx) and superoxide dismutase (Cu/ZnSOD) using the real-time PCR technique with LUX (Light Upon eXtension) fluorogenic primers. During the peripartum period (–22 days until + 22 days from calving), Cu/ ZnSOD mRNA expression decreased (p

This study investigated the morphometrical changes in mammary glands of 25 Sarda dairy ewes (5 per period) at 10 days before parturition (LateP); 30, 60, 150 days in milking (30, 60 and 150 DIM) and 8 days after the dry off (8 IN). Total number of alveoli and mammary epithelial cells (MEC) decreased during lactation, being the highest in LateP (2.54 × 106 and 60.6 × 106, respectively), intermediate at 30 DIM (1.36 × 106 and 27.7 × 106, respectively) and the lowest afterwards (0.56 × 106 and 12.3 × 106, respectively, at 8 IN). Number of MEC per alveolus (20.8 ± 1.58, mean ± SE) and estimated milk yield per MEC (3.8 × 10−5 ± 6.21 × 10−6) did not differ between periods. Mammary gland volume and internal morphology (number of MEC per alveolus, number of alveoli per udder half and volume of the alveolar lumen) explained, respectively, 62 and 47% of the variability of daily milk yield. The volume or weight of the mammary gland, which represent the black box in deterministic models, was more efficient in forecasting milk yield than the internal morphology of the udder, normally used in biomechanistic models.

The obese gene encodes leptin, a 16-kDa protein involved in the regulation of fat deposition and energy consumption.Backfat is one of the peculiar characteristics of Italian ham, and represents a fundamental quality factor. Therefore, theobese gene can be considered as a candidate marker for determining economically important production traits such asbackfat thickness, feed intake, and growth rate in swine. The aim was to investigate the relationship between obese genepolymorphisms and carcass and meat characteristics of pigs reared for ham production. In the present research, the analysesof three new RFLPs are reported. An MwoI polymorphism occurs at nucleotide 1792, within the intron. Pigs heterozygousat this position have heavier thighs with a thinner layer of fat. Two SmaI polymorphisms occur at nucleotides5018 and 5410 within the 3’ UTR of the obese gene. Animals heterozygous at position 5410 have characteristics suitablefor the production of San Daniele ham: lower backfat thickness and heavier thighs with a thinner fat layer, relative toother genotypes.

Randomly amplified polymorphic DNA (RAPD)-PCR was applied to the identification and typing of 218 Streptococcus thermophilus isolates from four Italian protected designation of origin type of cheeses. The pronounced differences among the RAPD-PCR profiles obtained with the primers M13 and D11344 revealed the presence of a significant genetic heterogeneity inside the species S. thermophilus. Randomly amplified polymorphic DNA-PCR also allowed to discriminate part of the S. thermophilus isolates according to the cheese origin. Finally, isolates showing RAPD-PCR profiles very similar to the profiles of two Streptococcus macedonicus reference strains were detected, suggesting the presence of these streptococci also in Italian cheeses.

Elena Pomari, Bruno Stefanon, Monica Colitti Department of Agricultural and Environmental Sciences, University of Udine, Udine, Italy Background: Arctium lappa (AL), Camellia sinensis (CS), Echinacea angustifolia, Eleutherococcus senticosus, Panax ginseng (PG), and Vaccinium myrtillus (VM) are plants traditionally used in many herbal formulations for the treatment of various conditions. Although they are well known and already studied for their anti-inflammatory properties, their effects on H2O2-stimulated macrophages are a novel area of study. Materials and methods: Cell viability was tested after treatment with increasing doses of H2O2 and/or plant extracts at different times of incubation to identify the optimal experimental conditions. The messenger (m)RNA expression of TNFα, COX2, IL1β, NFκB1, NFκB2, NOS2, NFE2L2, and PPARγ was analyzed in macrophages under H2O2 stimulation. The same genes were also quantified after plant extract treatment on cells pre-stimulated with H2O2. Results: A noncytotoxic dose (200 µM) of H2O2 induced active mRNA expression of COX2, IL1β, NFE2L2, NFκB1, NFκB2, NOS2, and TNFα, while PPARγ was depressed. The expression of all genes tested was significantly (P

Host and dietary interactions with the rumen microbiome can affect the efficacy of supplements, and their effect on the composition of the bacterial population is still unknown. A 16S rRNA metagenomic approach and Next-Generation Sequencing (NGS) technology were used to investigate the bacterial microbiome composition in the liquid fraction of the rumen content collected via stomach tubing. To investigate biodiversity, samples were taken from three groups of four lactating dairy cows given a supplement of either 50 g of potato protein (Ctrl group), or 50 g of lyophilized Saccharomyces cerevisiae (LY group) or 50 g of dried S. cerevisiae (DY group) in a potato protein support. Rumen samples were collected after 15 days of dietary treatments and milk production was similar between the three groups. Taxonomic distribution analysis revealed a prevalence of the Firmicutes phylum in all cows (79.76%) and a significantly (P

Neutrophils play an essential role in host defense and inflammation. In recent years, several studies in men and dairy animals have shown that administration of natural compounds can influence the immune system. In the present study the extracts of six patented protected plants, named C1, C2, C3, C4, C5, C6, have been screened for their effect on ovine neutrophils. Three, clinically healthy, non lactating, non pregnant sheep were used. Freshly isolated neutrophils were incubated with extracts at increasing doses and tested for viability, adhesion and superoxide production induced by PMA. The residues of C1, C2, C3 strongly inhibited adhesion and superoxide production, demonstrating anti-inflammatory activity. Further in vivo experiments are required to validate their use for productive animals.

Thirty five Duroc x Large White (DUxLW) and 43 Commercial Hybrid x Large White (CHxLW) were fed two diets differing for the content of crude protein (CP), with the aim to evaluate the effects of these diets on growth, carcass and thigh traits. Pigs were allotted to two groups and fed high protein (HP) and low protein (LP) diet. Within each dietary group, 3 diets were formulated, for the initial phase of growth (from 79 to 112 days, 17.3% and 15.4% CP as fed basis for the HP and LP diet, respectively), for the intermediated phase of growth (from 113 to 196 days, 15.1% and 13.7% CP as fed basis for the HP and LP diet, respectively) and for the finishing period (from 197 to 272 days, 13.4% and 11.4% CP as fed basis for the HP and LP diet, respectively). HP diets were supplemented with Lysine, LP were supplemented also with Methionine and Threonine. Pigs were slaughtered at nine months of age. Body weight (BW), average daily gain, total feed intake and feed conversion ratio were not affected by diet. Carcass weight, thigh weight, backfat and lean thickness and lean percentage (Fat-O-Meater, FOM) were not affected by dietary treatments, whilst backfat thickness and FOM were significantly higher (P

Pro-inflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor- alpha (TNF-α) play a key role in the pathogenesis of osteoarthritis (OA). Once released, these cytokines are potent stimulators for the de novo production of catabolic enzymes such as matrix metalloproteinases (MMPs) and cyclo-oxygenase-2 (COX-2). Anti-inflammatory agents capable of suppressing the production and catabolic actions of these cytokines may have therapeutic potential in the treatment of OA and a range of other osteoarticular disorders. The purpose of this study was to examine the therapeutic effect of Curcumin (diferuloylmethane), a pharmacologically safe phytochemical agent, on males and foals affected by degenerative joint diseases. Curcumin, in the form phytosome (CURCUVET ®, Indena Spa, Milan, Italy) was administered to animals for fifteen days and gene expression was monitored before the treatment and after four, eight, and fifteen days. In mares, Curcumin inhibited the expression of COX-2, TNF-α, IL-1β, IL1RN, and IL6, even if only the downregulation of IL-1β and IL1RN were significant. In foals, Curcumin significantly inhibited the expression of COX-2, TNF-α, IL1RN and significantly increased that of IL6. These results indicate that Curcumin has nutritional potential as a natural anti-inflammatory agent for treating osteoarticular disorders through suppression of pro-inflammatory cytokines and catabolic enzymes.

Permanent grassland of a typical Alpine region in Austria (Styria, 700 m above sea level, 1100 mm precipitation) was cut either 2, 3 or 4 times per year, conserved as barn-dried hay and fed to dairy cows at three concentrate levels [zero, concentrate according to requirements or 25% of dry matter (DM) intake concentrate]. Fertilisation levels were 32 m 3 slurry with or without 100 kg N ha −1 mineral N. Because of lower DM yield (8.65, 8.05, 6.51 t ha −1 ) and higher forage intake (10.4, 13.2 and 15.3 kg DM) the potential stocking rate decreased with increasing cutting frequency. Milk yield and N excretion per cow increased with increasing cutting frequency. When milk yield and N excretion were related to the forage area there was only a small influence of cutting frequency at the low level of concentrate. When the forage was supplemented with concentrate according to requirements, both milk yield and N excretion per ha decreased with increasing cutting frequency. Additional fertilisation of 100 kg mineral N increased the amount of slurry from 37.5 to 41.4 m 3 ha −1 forage and excretion of N from 149 to 160 kg ha −1 forage.

An investigation of the kinetics and synchronicity of rumen crude protein and starch degradability was performed for maize, flaked maize, ensiled maize cob, barley, flaked barley, wheat, oat, sorghum and triticale grain, using the in situ polyester bag technique. Kinetics of rumen degradability were corrected for particle losses from the bag before degradation. Washing losses were measured by shaking feed samples in polyester bags in tap water at 20°C for 1 h and recovering the particle losses in fibre glass filter. Mean washing losses of dry matter were 442 g/kg DM (soluble fraction 17%); mean washing losses of nitrogen were 446 g/kg nitrogen, with 52% being water soluble. Starch escaped to a higher extent from the bags in the washing machine (average washing losses were 581 g/kg starch, with a soluble fraction of 7%). Degradability data for dry matter, nitrogen and starch measured at each time of incubation were corrected for the respective particle losses and fitted with both first order and Gompertz (sigmoidal) models. The difference between the estimated parameters obtained with the two models was negligible, although differences occurred for the immediately soluble fraction (a) of dry matter and starch and for total degradable fractions of dry matter, nitrogen and starch. No differences were observed between effective degradabilities, independent of rumen outflow rate (0.04 and 0.08/h), apart for the mean effective degradability of nitrogen at 0.08/h, which was higher for the first order model (577 versus 564 g/kg, P

To compare growth, skeletal development, carcass traits and meat quality of different genotypes, 10 Casertana (CT), 10 Italian Large White (LW) and 10 Duroc x (Landrace x Italian Large White) (DU) crosses, barrows of 90-day-old, were allotted to the same outdoor rearing and feeding conditions. Live weight was recorded and average daily gain (ADG) was calculated. At slaughter (330-day-old) dressing and lean percentages were determined; backfat thickness and loin eye depth were measured. Carcasses were dissected into commercial cuts. Water holding capacity, pH and colour (45 min and 24 h post-mortem) were measured. Longissimus lumborum muscle samples were collected for cholesterol, α-tochopherol and intramuscular collagen (IMC) analyses. CT compared to DU and LW had the lowest growth rate and skeletal development. CT showed higher backfat thickness, lower lean cut/fatty cut ratio and less lean meat (P<0.05). Loin eye depth differed among genotypes with LW > DU > CT (P<0.05). CT showed higher red colour of the meat than DU and LW (P<0.05). CT compared to LW had the highest hydroxylysylpiridinoline (HLP) crosslink concentration and HLP/IMC ratio, and a lower IMC amount (P<0.05). CT pigs produced meat that could be tougher than that from the improved breed, but more acceptable from the technological point of view. At eleven months of age bone weight, length and diameter were clearly genetic type-related; differently, the bone maturity was similar among the genotypes studied.

The purpose of the study was to evaluate associations between average daily gain (ADG) and DNA variability in the regulatory region of candidate genes in Italian Large White, Italian Duroc and Italian Landrace breeds for heavy pig production. The ADG of pigs was available as Estimated Breeding Values (EBVs) and Random Residuals (RRs). Within each breed, 200 individuals were sampled, 100 in the higher and 100 in the lower 5% tail of the normal distribution curve according to the EBVs for ADG of the populations (600 pigs overall). Six SNPs in promoter regions of 5 candidate genes (MYF6, MYOD1, OPN, SCD and two for LEP) were analysed. Allele frequencies were calculated and the SNPs for LEP, MYF6 and MYOD1 segregated in the 3 breeds, while polymorphisms in the OPN and SCD genes did not segregate in Italian Large White and Italian Landrace, and Italian Landrace respectively. Fisher’s exact test (two-tailed) for the distribution of allele frequencies in minus and plus variant groups was significant for LEP_1 and MYOD1 in Italian Large White, for LEP_1, LEP_2, MYOD1, OPN and SCD in Italian Duroc and for LEP_1, LEP_2 and MYOD1 in Italian Landrace. Several putative binding sites were detected in the promoter regions using MATINSPECTOR software. Analysed mutations may affect putative core sequences of binding sites for transcription factors involved in muscle and fat tissues deposition. However, variations in binding affinities of the sequences related to the SNPs and the expected differential gene expression require to be evaluated.

Three experimental diets were formulated to contain the same amount of fescue hay (560 g kg−1 dry matter (DM) Festuca arundinacea) and differing condensed rennet whey:maize meal ratios (0:100, 50:50 and 100:0 on a DM basis, diets M, MW and W, respectively). The diets were fed during two experimental periods to six rumen fistulated, non lactating Simmental cows (average level of intake: 77 g DM kg−1 LW0.75, four cows per treatment). Whey used alone caused a lower rumen pH at 1.5 and 3.0 h after the morning meal (P < 0.01) in comparison with the other two diets, while after 4.5 h rumen acidity values were similar between diets. After feeding, the inclusion of whey led to regular increases (P < 0.05) in the concentrations of propionic acid at 1.5 h (12.4 mmol, 18.3 mmol and 23.8 mmol l−1, respectively for M, MW and W diets), while at 3.0 h the W diet remained significantly higher than the other two (22.1 mmol vs. 14–14.2 mmol l−1, P < 0.01). This was also the case for butyric acid where higher (P < 0.01, 1.5 h) levels were observed for diets containing whey (11.6 and 13.3 mmol vs. 7.9 mmol l−1). The dry and organic matter in the W and MW diets were 3 percentage points (P < 0.05) more digestible than the M diet. The soluble cell content fraction digestibility regularly and significantly increased (P < 0.05) as whey substituted maize (from 749 to 814 g kg−1). The inclusion of whey caused an increase in rumen liquid turnover rate (from 7.1 to 13.4–12.3% h−1, P = 0.058) and enhanced levels of urinary volume which almost doubled in diets containing whey (from 78 to 132–138 mg kg−1 LW0.75, P < 0.05). Urinary allantoin output did not differ between dietary treatments while higher urinary outputs of uric acid were found for cows fed diets containing whey (2.39–2.58 g vs. 1.36 g day−1, P < 0.05).

The aim of the present research was to investigate the regulation of gene expression in ovine blood leukocytes during ACTH-induced cortisol release and the effect of dietary administration of botanicals to counteract the evoked response in polymorphonucleate cells (PMNCs). Thirty-six homogeneous Sarda sheep (age, 18 ± 4.1 mo; BW, 38.7 ± 1.3 kg) were allotted to six groups of six sheep each. One group was used as a negative control (Saline) and five groups were treated, every 12 h for 48 h, with 0.5 mL of ACTH agonist (250 μg/mL of tetracosactrin). Before ACTH treatment, four of the five ACTH-treated groups were separated and fed for 22 d with a basal diet supplemented with extracts from Echinacea angustifolia roots (PO + ACTH), Echinacea angustifolia flowers (EA + ACTH), Andrographis paniculata (AP + ACTH), and the bark of Larix decidua milled (LB + ACTH). Control groups (Saline and ACTH) were fed with the same basal diet without botanicals. Total RNA was extracted from blood samples collected before (T0) and after 3 h (T3) and 51 h (T51) from the first ACTH injection, and transcriptome analysis was performed using a custom oligoarray, designed from 12,194 Ovis aries UniGenes on a CombiMatrix platform. At T3, treatment with ACTH caused down-regulation of transcripts (P < 0.001) involved in "response to stress" (GADD45A, GADD45B, WRNIP1, and XRCC6) and in "innate immune response" (MAPK3 and NFkBIB). At T51, treatment with ACTH caused down-regulation (P < 0.001) of genes involved in "immune response" (IFNG and IL2) and up-regulation (P < 0.001) of NF-κB1 and TP53. Each botanical produced a different (P < 0.001) molecular signature for these genes at T3 and T51. The most active botanical in modulating transcriptome modifications in PMNCs after ACTH-induced cortisol release was Larix decidua Mill bark followed by Polinacea roots. These botanicals can be viewed as promising feed supplements in ruminants to cope with conditions associated with increased concentrations of plasma cortisol