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ABSTRACT: Costs of production have deeply increased each year in the last decades, breeders are continuously looking for more cost effective and more efficient ways to produce milk. Despite the major signs of progress in productivity, it is fundamental to optimize rather than maximize the performances of the dairy cows. Mastitis is still a highly prevalent disease in the dairy sector which causes several economic losses and environmental effect. Its accurate and early diagnosis is crucial to improve profitability of dairy cows and contribute to a more sustainable dairy industry. Among mastitis reduction strategies, there is the urgent need to implement breeding objectives to select cows displaying mastitis resistance by investigating the genetic mechanisms at the base of the inflammatory response. Therefore, in this study we aimed to further understand the genetic background of the differential somatic cell count (DSCC), which provides thorough insights on the actual inflammatory status of the mammary glands. The objectives of this study were to estimate on a cohort of 20,215 Italian Simmental cows over a 3-yr period: (1) the heritability and repeatability values of somatic cell score (SCS) and DSCC, (2) the genetic and phenotypic correlations between these 2 traits and milk production and milk composition traits, (3) the heritability and repeatability values of SCS and DSCC within class of udder health status. Heritability was low both for SCS (0.06) and DSCC (0.08), whereas the repeatability values for these traits were 0.43 and 0.36, suggesting that the magnitude of cow permanent environmental effect for these traits is remarkable. The genetic and phenotypic correlation of SCS with DSCC was 0.612 and 0.605, respectively. Because both significantly differed from the unit, we must consider those traits as different ones. This latter aspect corroborates the need to consider the DSCC as a further indicator of inflammatory status which might be implemented in the Simmental breed genetic evaluation. It is worthy to mention that heritability estimates for SCS and DSCC were the highest in healthy cows compared with the other udder health classes. This implies that when the udder health status changes, it is most likely due to environmental factors rather than aspects related to the animal's genetics. In contrast, the highest additive genetic variance and heritability found for SCS and DSCC in the healthy group might reveal the potential to further implement breeding strategies to select for healthier animals.

This study aimed to determine the impact of a novel formulation of a supplement composed of the natural ingredients, bromelain, quercetin, and Lentinula edodes, on the gut microbiota of healthy adult dogs. Adult healthy female dogs were administered either a placebo (CTR, n = 15) or the supplement (TRT, n = 15) over 28 days. Stool samples were collected for 16S rRNA sequencing before supplement administration (T0), at completion of supplement administration (T28), and one week after the end of supplement administration (T35) to characterize changes in the gut microbial communities. QIIME was used to determine both alpha- and beta-diversity, and ANCOM-BC was used to identify differences in taxonomic abundances before and after supplementation. We found a significant decrease in overall diversity in the CTR group but no significant differences in overall diversity in the TRT group over time. Furthermore, we found differences in the abundance of several taxa in both the CTR and TRT groups, but differences in the abundance of beneficial bacteria were more pronounced in the TRT group. Specifically, we found increases in the abundance of sequences belonging to the genera Bifidobacterium, Lactobacillus, and Pediococcus at T28 in the TRT group with significant increases in Bifidobacterium and Lactobacillus persisting at T35 when compared to T0. Importantly, members of these genera are considered important for their anti-inflammatory properties, vital for fostering a balanced and robust gut microbiota in dogs. The results of our study show the potential of our supplement to selectively enhance specific beneficial bacterial taxa, offering a targeted approach to modulating the gut microbiome without causing disruptions to the overall equilibrium.

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A growing number of studies in the last decade described the microbial populations in different niches of the organism thank to High-throughput DNA sequencing techniques, that are easily accessible to researchers. Furthermore, network analysis allows characterisation of bacteria that are indirectly linked to outcomes such as disease, diet and host sex through their association with other taxa. The present work on the gut microbiome follows previous research in which data were collected from 8 in-house dietary intervention studies in healthy dogs. Animals were divided according to diet and sex. The process of sample collection, storage, DNA extraction and sequencing, bioinformatic and statistical analysis followed a tailored, internally defined pipeline. The extracted DNA was prepared for sequencing of the V3 and V4 regions of the 16 rRNA gene. Correlation network analysis was performed by calculating pairwise relationships between taxa using the Sparse Correlations for Compositional Data (SparCC) algorithm. First, we identified candidate bacteria that were highly abundant in the microbial community, and second, we examined taxa that were directly related to diet and sex factors. In summary, this study underpins the network structure of the gut microbiome of dogs categorised by diet and sex and provides a better explanation for the interactions between bacteria that led to the grouping of dogs based on environmental or genetic factors. Specifically, this study has provided the basis for understanding how the bacterial community in the gut is interconnected and functions as a function of diet composition and sex.Highlights Network analysis could allow indirect association between bacteria composing the gut microbiome and outcomes, such as disease, diet and sex of the host. This research put the basis on the definition of enterotypes also in dogs, a concept already applied in humans. Positively correlated bacteria, belonging to modules could help to modulate diets and therapies for the treatment of enteropathies or gut-related pathologies linked to a dysbiosis status of the host.

Overweight and obese dogs can develop metabolic dysfunction, characterized by an inflammatory response and involvement of liver functions. If a modulation of the gut microbiome and its interaction with the gut–liver axis is implicated in the development of metabolic dysfunction, exploration becomes necessary. Over the past decade, diverse therapeutic approaches have emerged to target pathogenic factors involved in metabolic dysfunction. This study investigated the impact of a supplement with hepatoprotective activity, containing extracts of Silybum marianum, prebiotics, probiotics, n-3 polyunsaturated fatty acids, vitamins, and minerals on hematological markers of liver functions and inflammation, as well as on the intestinal microbiota of 10 overweight adult dogs over a 35-day time span. Animals underwent clinical and laboratory evaluations every 7 days, both before the administration of the supplement (T0) and after 7, 14, 21, 28, and 35 days (T1, T2, T3, T4, and T5). In comparison to T0, a significant (p < 0.05) decrease in ALP, glucose, direct bilirubin, and CRP was observed from T3 to T5. The alpha diversity of the fecal microbiota significantly decreased (p < 0.05) only at T1, with high variability observed between dogs. Total short-chain fatty acid and lactic acid were also lower at T1 (p < 0.05) compared to the other times of sampling. The beta diversity of the fecal microbiota failed to show a clear pattern in relation to the sampling times. These results of blood parameters in overweight dogs show a reduction of the inflammation and an improvement of metabolic status during the study period, but the effective contribution of the supplement in this clinical outcome deserves further investigation. Furthermore, the considerable individual variability observed in the microbiome hinders the confident detection of supplement effects.

Oxidative stress causes several pathological conditions in humans and animals, including gastrointestinal disorders. The aim of this study was to analyze the antioxidant capacity of three natural powdered raw materials containing quercetin, bromelain, and Lentinula edodes and develop a new feed supplement for dogs using a combination of them. The total phenolic content (TPC), antioxidant activity, DPPH (2,2-diphenyl-1-picrylhydrazyl), and ABTS (2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) of the extracts, either individually or in combination, were evaluated colorimetrically. The effects of this supplement on healthy adult dogs’ nutritional, inflammatory, and stress status were evaluated. American Staffordshire Terrier adult female dogs (n = 30) were randomly assigned to a control (n = 15) or a treated (n = 15) group. The supplement was added as powder to the food of the treated dogs once daily for 28 days. There was no significant difference in the body weight and body condition scores between the initial and final phases of the experiment. At the end of our study, a significant decrease in fecal calprotectin, cortisol, indole/skatole, and N-methylhistamine and a significant increase in short-chain fatty acids were observed as compared to the control group. In conclusion, this natural feed supplement can be used to improve gastrointestinal health and psycho-physical conditions in dogs.

Recent studies have found bacterial DNA in the blood of healthy individuals. To date, most studies on the blood microbiome have focused on human health, but this topic is an expanding research area in animal health as well. This study aims to characterize the blood microbiome of both healthy dogs and those with chronic gastro-enteropathies. For this study, blood and fecal samples were collected from 18 healthy and 19 sick subjects, DNA was extracted through commercial kits, and the V3-V4 regions of the 16S rRNA gene were sequenced on the Illumina platform. The sequences were analyzed for taxonomic annotation and statistical analysis. Alpha and beta diversities of fecal microbiome were significantly different between the two groups of dogs. Principal coordinates analysis revealed that healthy and sick subjects were significantly clustered for both blood and fecal microbiome samples. Moreover, bacterial translocation from the gut to the bloodstream has been suggested because of found shared taxa. Further studies are needed to determine the origin of the blood microbiome and the bacteria viability. The characterization of a blood core microbiome in healthy dogs has potential for use as a diagnostic tool to monitor for the development of gastro-intestinal disease.

Metabolomics provides a description of the phenotype of an organism and complementary biochemical information to genomics and proteomics. The purpose of this research was to depict the metabolite profile of faecal samples from dogs fed three different diets through NMR spectroscopy analysis. Samples were collected from 14 dogs fed a commercial extruded diet, 18 dogs fed a homemade diet and 16 dogs fed a raw meat-based diet. The average BCS for all dogs was 4–5 and the average Faecal Score was 2–3. Only healthy animals were considered, as assessed from the clinical evaluation of the veterinarians. Faecal samples were prepared using phosphate buffer (pH 7.1) combined with deuterated water and analysed with NMR spectroscopy using a Bruker Avance III HD 400 MHz spectrometer. Principal component analysis of the spectra signals demonstrated clustering of dogs according to diet, with 57.8% of the variance explained by the first three components. Targeted metabolome analysis was also performed on 56 metabolites of interest, selected from a database of 558 metabolites. Our data suggest that metabolome analysis using NMR is a promising approach to describe the phenotypic variation that occurs among dogs fed different diets.The metabolomic approach is becoming always more important in the evaluation of the functional responses of animals to dietary intervention. NMR-metabolomics has emerged as a non-destructive technique that allows a simpler data interpretation compared to other techniques. NMR metabolome analysis resulted to be a promising approach to describe the phenotypic variation that occurs among dogs fed different diets.

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

The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of polymorphonuclear cells, 10 cows were assigned to group H, 11 to group ARM, and 11 to group SCM. After isolating exosomes in milk by isoelectric precipitation and ultracentrifugation, the extracted RNA was sequenced to 50 bp long single reads, and these were mapped against Btau_5.0.1. The resulting 225 miRNAs were uploaded to the miRNet suite, and target genes for Bos taurus were identified based on the miRTarBase and miRanda databases. The list of differentially expressed target genes resulting from the comparisons of the three groups was enriched using the Function Explorer of the Kyoto Encyclopedia of Genes and Genomes. A total of 38, 18, and 12 miRNAs were differentially expressed (DE, p < 0.05) in the comparisons of H vs. ARM, ARM vs. SCM, and H vs. SCM, respectively. Only 1 DE miRNA was shared among the three groups (bta-mir-221), 1 DE miRNA in the H vs. SCM comparison, 9 DE miRNAs in the ARM vs. SCM comparison, and 21 DE miRNAs in the H vs. ARM comparison. A comparison of the enriched pathways of target genes from the H, SCM, and ARM samples showed that 19 pathways were differentially expressed in the three groups, while 56 were expressed in the H vs. SCM comparison and 57 in the H vs. ARM comparison. Analyzing milk exosome miRNA cargos can be considered as a promising approach to study the complex molecular machinery set in motion in response to mastitis in dairy cows.

The review described the most important factors affecting the development of the intestinal microbiota in puppies from birth to weaning. The health and well-being of the microbiome in puppies is influenced by the type of parturition, the maternal microbiota, and the diet of the mother, directly or indirectly. The isolation of bacteria in dogs from the placenta, fetal fluids, and fetuses suggests that colonization could occur before birth, although this is still a matter of debate. Accordingly, newborn puppies could harbor bacteria that could be of maternal origin and that could influence microbial colonization later in life. However, the long-term impacts on health and the clinical significance of this transfer is not yet clear and needs to be investigated. The same maternal bacteria were found in puppies that were born vaginally and in those delivered via cesarean section. Potentially, the relationship between the type of parturition and the colonization of the microbiome will influence the occurrence of diseases, since it can modulate the gut microbiome during early life. In addition, puppies’ gut microbiota becomes progressively more similar to adult dogs at weaning, as a consequence of the transition from milk to solid food that works together with behavioral factors. A number of researches have investigated the effects of diet on the gut microbiota of dogs, revealing that dietary interference may affect the microbial composition and activity through the production of short-chain fatty acids and vitamins. These compounds play a fundamental role during the development of the fetus and the initial growth of the puppy. The composition of the diet fed during pregnancy to the bitches is also an important factor to consider for the health of newborns. As far as it is known, the effects of the type of parturition, the maternal microbiota, and the diet on the microbial colonization and the long-term health of the dogs deserve further studies. Definitely, longitudinal studies with a larger number of dogs will be required to assess a causal link between microbiome composition in puppies and diseases in adult dogs.

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.

The aim of this study was to evaluate the effect of the administration of Saccharomyces boulardii on the nutritional, immunological, inflammatory, and stress status and on the composition of the gut microbiota and mycobiota in healthy adult dogs. A total of 25 American Staffordshire Terrier dogs were selected and randomly assigned to two groups: control (CTR, n = 12) and treated (TRT, n = 13) groups. No significant differences were found between the two groups regarding body weight, body condition score, and fecal score. No significant differences in microbiota/mycobiota, short chain fatty acids, indole/skatole, histamine, zonulin, or lactoferrin were detected. Indeed, supplementation with S. boulardii significantly decreased fecal calprotectin Immunoglobulin A, indicating an improvement in the gut well-being. Interestingly, fecal cortisol significantly decreased in dogs belonging to the TRT group compared to the CTR, suggesting both an improvement of the intestinal status and a reduction of stress, a common condition affecting animals managed in a breeding environment.

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.

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

Working dogs are gaining popularity for their ability to learn and perform tasks entertaining their human companions. For this reason, dogs are often subjected to various stimuli due to inter- and intra-specific interactions, environmental variations and effort required by different activities. In the present study, salivary cortisol was measured to monitor physiological response to different conditions. The first study was performed to assess the variability of salivary cortisol in dogs in usual environmental conditions. For this, salivary cortisol was measured in 10 dogs at home during three not consecutive days at three different times of the day and not significant variations between days and time of sampling were observed. In the second study, salivary cortisol was measured in dogs before and after Pointing Hunting (No. 5), Tracking for Ungulate Hunting (No. 6), Blood Tracking (No. 4), Agility Training (No. 6) and Animal Assisted Activities (AAA, No. 6). Salivary cortisol concentration significantly increased after the Pointing Hunting activity (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.

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 < 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.

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.

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.

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.

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.

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.

Differential leukocyte count (DSCC) in milk is considered important to improve knowledge of udder immune response. The investigations on milk DSCC were limited by the techniques available until recently, when a high-throughput tool to perform DSCC opened the way to explore these factors in rapid and economically sustainable ways. We hypothesized that DSCC alone does not fully describe the pattern of these cells, since the total amount is also influenced by milk yield and SCC. Therefore, this study was designed to describe DSCC and total amount of different leukocytes in milk during the course of lactation in cows differing in parity and in levels of SCC. This study considered 17,939 individual milk tests from 12 dairy herds in Lombardy Region, where DCC testing was applied in the period of February 2018–December 2019 (23 months). The samples were divided into two subsets—“healthy” (HS) with SCC ≤200,000 cells/mL and “inflamed” (IS) with SCC >200,000 cells/mL. Cow in HS have a P + LT average between 5.0 × 108 and 3.0 × 109 cells. In IS cows, the values were 1.6 × 1010 and 2.5 × 1010. Therefore, the presence of a well-defined inflammatory process increased the overall amount of polymorphonuclear neutrophils (PMN) and lymphocytes (LYM) of 1 log, from 1 × 109 to 1 × 1010. The assessment of the total amount of PMN and LYM, to our knowledge, have never been reported in scientific literature; the values observed may be proposed as benchmarks for studies on udder immune response. When data were analyzed by days in milk (DIM), they showed that cows in first and second lactation have a significantly lower amount of PMN + LYM, when compared to cows in third and higher lactation. However, these differences are numerically not very large (7%), and suggest that, in healthy animals, the number of immune cells is kept as constant as possible. In IS, the analysis of trends based on DIM showed that both DSCC and P + LT have a significant negative trend. These data suggest that only in this group, the presence of high SCC as lactation proceeds is associated with a progressive increase in the number of macrophages. To the best of our knowledge, this is the first study describing the pattern of DSCC and the total amount of PMN + LYM in relation to parity, days in milk, and SCC, and it may be considered as the first contribution in the investigation on mammary gland immune response by the means of differential cell counts in milk.

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 < 0.01) of Enterococcus and Adlercreutzia were observed in D0, whilst Escherichia and Eubacterium were higher in D1. Fusobacterium and Phascolarctobacterium were higher (p < 0.01) in D3. Salivary serotonin increased (p < 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.

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.

In the veterinary sector, many papers deal with the relationships between inflammation and oxidative stress. However, few studies investigate the mechanisms of action of oxidised molecules in the regulation of immune cells. Thus, authors often assume that these events, sometime leading to oxidative stress, are conserved among species. The aim of this review is to draw the state-of-the-art of the current knowledge about the role of oxidised molecules and dietary antioxidant compounds in the regulation of the immune cell functions and suggest some perspectives for future investigations in animals of veterinary interest.

Skeletal muscle growth represents one of the main economic traits in pig production. To gain a better understanding of expressions profile in pig muscle, serial analysis of gene expression (SAGE) was performed in Longissimus dorsi of two pigs at 3 and 9 months of age. A total of 53,120 long tags were obtained and sequenced from the four muscle SAGE libraries, representing 17,902 different tags, or putative transcripts, 0.64% (+0.09) of which had a relative expression level higher than 1‰. Overall, a total of 218 tags were highly expressed and 31 had a frequency higher than 3‰. Functional characterisation of the expression profiles was performed using Kyoto Encyclopedia of Genes and Genomes metabolic maps and 139 pathways were identified for swine skeletal muscle. Focal adhesion, Mitogen-Activated Protein Kinase signalling, oxidative phosphorylation, ribosomal proteins, regulation of actin cytoskeleton and insulin signalling pathways showed an abundance of genes greater than 1.5% of all the expressed transcripts. A comparison with human SAGE library indicated no statistical differences for the frequency of genes involved in these metabolic pathways.

The aim of this research was to evaluate the effect of the modification of diet starch content on milk composition and onits nitrogen and mineral fractions. Ten Italian Friesian primiparous cows were randomly assigned to two groups and feda basal total mixed ration, (BSD, basal starch diet, 24.9% starch/DM) until 42 days in milking (DIM). At 43 DIM, 5 animals(control group, CTR) continued to receive the same ration and the remaining 5 cows (experimental group, EXP)were fed a low starch diet (LSD, 21.0% starch/DM) until 65 DIM, followed by a high starch diet (HSD, 28.3% starch/DM)66 to 85 DIM. From 86 DIM until 94 DIM, cows of the EXP group returned to the BSD. Milk samples were collected at 37,50, 60, 70, 80, 94 DIM. Starch intake was lower for EXP at DIM 50, 60 and higher at 70 and 80 DIM (P < 0.01). Milkyield and fat corrected milk (FCM) did not vary between groups and times of sampling, but HSD caused a significant (P< 0.05) reduction of milk fat in the EXP animals. NPN and MUN contents were affected by dietary treatments at 80 DIM(P < 0.05), being higher in CTR and lower in EXP group in comparison to their basal values (37 DIM). The αs1-caseinpercentages at 80 and 94 DIM increased in the EXP but not in the CTR group, and a marked decrease of αs2-casein percentagefor the EXP group at 94 DIM was observed (P < 0.05). Milk sodium content decreased at 80 and 94 DIM and thesodium to potassium ratio was reduced after the return to the basal diet in the EXP group (94 DIM), indicating that dietarystarch variations can be involved in the control of epithelium integrity of mammary gland in early lactation.

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.

The aim of the research was to compare gene transcription profiles of Musculus longissimus dorsi (MLD) between pigs fed diets with high (HED) or low (LED) energy contents. Two groups of 4 Casertana pigs were reared from 3 to 12 months of age in the same environmental conditions and fed HED or LED. In the HED, the ave rage daily gain and back fat thickness were significantly (P

The volatile fractions of milk and their evolution in ripened cheese were studied using purge and trap extraction and gas chromatography-mass spectrometry analysis (GC-MS). Eighteen samples of raw milk and cheese ripened for 70 days were collected in 2 consecutive years from dairy farms located in the border area between Italy and Slovenia. Twenty one volatile compounds not detected in the fresh milk used for cheese manufacturing, belonging mainly to the class of esters and alcohols, were found, while 8 compounds detected in milk were not found in cheese after 70 days of ripening. In cheese the majority of compounds increased significantly, even though the variations differed between the classes of volatile compounds. Concentration of alcohols, in particular ethanol, 1-propanol, 2-propanol, 3-methyl-1-butanol and 2-pentanol, increased considerably, while the content of ketones and terpenes was characterized by a significant, although small, rise that was largely related to a quantitative variation of only few compounds. Also esters, despite the high number of volatile compounds of new formation, showed a lower increase of concentration than that observed for the alcohols. Concentration of aldehydes, hydrocarbons and sulphur compounds remained unchanged, despite relevant variations involving the individual chemical compounds. Significant differences were also observed between the dairy product of the two countries, mainly for the concentration of volatile compounds belonging to the classes of alcohols, ketones and esters.

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.

Six batches of four commercial hybrids of heavy pigs, reared for the production of Italian dry-cured hams, were identifiedfor having homogeneous feeding and farm conditions. For a total of 235 pigs, slaughtered in the same slaughterhouse,carcass traits and muscle composition were measured. The pigs were genotyped for single nucleotide polymorphisms(SNPs) of Na+, K+-ATPase subunit alpha 2 (ATPase, Na+/K+ transporting, alpha 2 (+) polypeptide; ATP1A2), cystatinB (CSTB), mitochondrial 2,4-dienoyl-CoA reductase 1 (DECR1), leptin (LEP; 3 SNPs), melanocortin receptor 4(MC4R), melanocortin receptor 5 (MC5R), sarcolipin (SLN) and titin (TTN) genes. All genes showed biallelic polymorphismsand the alleles were differently distributed between the six batches. Pigs were subsequentely clustered in “lean”and “fat” using either carcass traits (lean percentage, backfat thickness, loin muscle thickness, ham weight and hamcover fat thickness: 100 lean and 135 fat) or meat composition data (dry matter, protein, fat and ash of Biceps femorisand Vastus lateralis and pH after 24 hours: 126 lean and 109 fat). The association of gene polymorphisms with leanessand fatness of pigs was thus investigated using a logistic regression. ATP1A2, LEP (HinfI polymorphism) and MC4R,together with sex and ham weight were, included in the model to screen lean and fat pigs classified according to carcasstraits data, yielding a correct classification of 71%. For the lean and fat pigs classified according to muscle composition,sex, CSTB, DECR1, MC5R and LEP (AciI/TaqI polymorphisms) were included in the regression analysis, that yielded a66% of pigs correctly classified.These preliminary results may indicate that some of the selected candidate genes could be associated to production traitsand are worth of further investigations.

The study investigated the relationship of markers of welfare and metabolism in milk, urine and blood with the genetic merit of Holstein and Simmental cows after the peak of lactation. Cows were selected from 3 Simmental (IS) and 2 Holstein (IH) commercial dairy farms. Within each farm, cows were ranked according to the estimated breeding value for milk protein yield (EBVp) from minus to positive and selected every 5 EBVps from minus to positive values (about 20% lactating cows for each farm). Milk was sampled and analyzed for protein, fat, lactose, cortisol contents and somatic cell count (SCC). Blood and urines were analyzed for biomarkers of metabolism and welfare. Significantly lower body condition score (BCS) was observed for IH in comparison to IS. Plasma creatinine was higher in IS, whilst Zn, total antioxidant status and glutathione peroxidase was higher in IH. The creatinine N to N ratio in urine was significantly higher for IS, while the purine derivatives (PD) N to creatinine N ratio was higher for IH. The EBVp was negatively related to BCS and glucose for IS and to plasma b-hydroxybutyrate in both breeds. EBVp was negatively related to urinary PD N to total N ratios for IS and to PD N to creatinine N ratio for IH. These preliminary results would indicate that the selection of cows for milk protein yield had minor effect on plasma and milk biomarkers of welfare. Instead, biomarkers of metabolism were more affected by breed and genetic merit.

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.

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.

The effect of Larix decidua (LD) dietary administration on gene expression patterns has been evaluated in sheep under ACTH challenge. Experimental protocol was approved by local laws and regulations. Eighteen sheep at maintenance were allotted to 3 groups: CTR (negative control, without ACTH, and supplementation), ACTH (positive control, with ACTH, and without supplementation), LD (ACTH and 50 g/head/day of LD). ACTH was injected for 3 subsequent days to ACTH and LD groups and blood was sampled before (T0) and after 3 (T3) and 51 (T51) hours from the first injection. RNA extracted samples were pooled together within group and time of sampling. A custom oligoarray was synthesized using 24,384 35 -40mer probes designed from 12,194 UniGenes (NCBI) on a CombiMatrix 90K platform. Cy5 labelled samples were hybridized on the chip. Statistical analysis, performed with MeV software 4.1 (TIGR), allowed the identification of a set of genes which were up or down regulated as a consequence of ACTH treatment. Genes that resulted differentially expressed were annotated with HomoloGene system and data mining was performed with Babelomics v3.1 tool. Functional analysis showed that most of the differentially expressed genes belong to KEGG pathways involved in immune system response and signaling molecules and interaction. Larch administration was effective in counteracting the effect of ACTH injection on the inflammatory processes, restoring the physiological homeostasis.

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

This review summarizes the importance of enteric methane (CH4) emission in ruminants and relevant to the current on knowledge relevant to genetic aspects of enteric CH4 production, highlighting future research needs and directions. Global average temperature has increased by about 0.7°C in the last century. The Intergovernmental Panel on Climate Change (IPCC) reported that anthropogenic greenhouse gases (GHG), including carbon dioxide (CO2), CH4, nitrous oxide (N2O) and halocarbons, have been responsible for most of the observed temperature increase since the middle of the twentieth century. Agriculture, particularly livestock, is increasingly being recognized as both a contributor to the process and a potential victim of it. Policy interventions and technical solutions are required to address both the impact of livestock production on climate change and the effects of climate change on livestock production. Food and Agriculture Organization (FAO), declared that in the next 50 years, the world’s farmers will be called upon to produce more food than has been produced in the past 10,000 years, and to do so in environmentally sustainable ways. Therefore, the GHG reduction should be treated as a public good. The United States congress is prospecting to define a price on GHG emissions. Limiting the concentration of CO2 and other GHG in Earth’s atmosphere requires a technological and economic revolution. A cost-effective way could be the genetic improvement of livestock, which produces permanent and cumulative changes in performance. Animal variation in enteric CH4 emission has been reported in the literature, providing potential for improvement through genetic selection.

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.

A dramatic increase of the demand for products of animal origin is expected to occur due to: i) the exponential growth of world’s population, which should reach 9 billion people in 2050, according to FAO predictions, ii) the process of urbanization which will likely lead to a concentration of 60% of the population in urban areas by the same year, and iii) the increase of income of a large part of the population in emergent countries such as China and India. So far, the increased demand for food has been supplied by agriculture due to improvement of techniques, increase of cultivated land areas and increase of consumption of water and energy. Despite that, unequal food distribution throughout the world has lead to almost 1 billion undernourished or starving people at the first decade of the twenty-one century. In the future, it will be necessary to achieve a sustainable supply of food, especially of animal origin, because land and other production factors are not unlimited resources. This work deals with related problems and possible sustainable solutions for the increasing demand for products of animal origin, by a detailed analysis of the Animal footprint, i.e. the Ecological footprint of animal production.

In 100 dried samples of forages, harvested three or four times during 1997 and 1999 in different morphological stages the macro- (Ca, P, Mg, K and Na) and trace- (Zn, Mn and Se) mineral contents were determined by chemical methods and by near infrared reflectance spectroscopy (NIRS). The accuracy calibration equations of NIRS for macrominerals Ca, P, Mg and K showed a high degree of accuracy, with an R2 of 0.963, 0.884, 0.892 and 0.939, respectively. These results suggest that these macrominerals are probably closely associated with the organic components, such as plant cell wall (Ca and Mg), chlorophyll (Mg), phytates, phospholipids, phosphoproteins and nucleic acids (P) and organic acids, such as malate (K). On the contrary, NIRS did not estimate Na content very accurately (R2 = 0.52; coefficient of variability of cross validation (CVSECV) = 35.5%), suggesting that Na is not closely associated with organic functional groups or that the amount of the mineral is below the level of NIRS detection. Similar conlusions are also valid for trace minerals, where the R2 was 0.642 (SECV = 4.4) for Zn, 0.427 (SECV = 55.6) for Mg and only 0.186 (SECV = 6.5) for Se. The CVSECV for trace minerals always exceeded 20%. Despite the insensibility of NIRS to accurately determine Na and certain trace minerals, the NIRS technique may provide an acceptable prediction of macro mineral contents.

One hundred dried samples of forage, harvested three or four times during 1997 and 1999 in different morphological stages were analysed for chemical composition, in vitro dry matter digestibility (IVDMD; Tilley and Terry, 1963) and in vitro gas production (Menke and Steingass, 1988) by laboratory methods and by near infrared reflectance spectroscopy (NIRS). NIRS predicted the main chemical components, such as crude protein, crude fibre, neutral detergent fibre, acid detergent fibre, with a very high degree of accuracy (i.e. the determination coefficient (R2) varied from 93.5 % for ADF to 98,2 % for NDF), while the coefficients of determination of crude ash, ether extract and acid detergent lignin were 79.6, 85.1 and 82.3 %, respectively, with standard errors of cross validation (SECV) exceeding 9.0 %. The IVDMD was also determined with a high degree of accuracy (R2 = 94.3 %; SECV = 4.3 %), such as parameters of gas production "B" (total gas production) and "A" (constant factor describing the decay in specific gas production rate), estimated with the Gompertz model, which have R2 and SECV values of 93.0 and 3.8 % and 94.5 and 8.0 %, respectively. The parameter "C" (the specific fermentation rate) was predicted with a somewhat lower degree of accuracy (R2 = 78.3 and SECV = 9.6 %).