Identification of ROH Islands Conserved through Generations in Pigs Belonging to the Nero Lucano Breed.

The recovery of Nero Lucano (NL) pigs in the Basilicata region (Southern Italy) started in 2001 with the collaboration of several public authorities in order to preserve native breeds that can play a significant economic role both due to their remarkable ability to adapt to difficult environments and the value of typical products from their area of origin. In this study, by using the Illumina Porcine SNP60 BeadChip, we compared the genetic structures of NL pigs reared in a single farm in two different periods separated by a time interval corresponding to at least three generations. The results showed an increase in the percentage of polymorphic loci, a decrease in the inbreeding coefficient calculated according to ROH genome coverage (FROH), a reduction in the number of ROH longer than 16 Mb and an increase in ROH with a length between 2 and 4 Mb, highlighting a picture of improved genetic variability. In addition, ROH island analysis in the two groups allowed us to identify five conserved regions, located on chromosomes 1, 4, 8, 14 and 15, containing genes involved in biological processes affecting immune response, reproduction and production traits. Only the conserved ROH island on chromosome 14 contains markers which, according to the literature, are associated with QTLs affecting thoracic vertebra number, teat number, gestation length, age at puberty and mean platelet volume.

Polymorphism at the CSN1S1 Locus and Energy Intake Level Affect Milk Traits and Casein Profiles in Rossa Mediterranea Goats.

A total of twenty-seven Rossa Mediterranea lactating goats, consisting of nine homozygous for strong alleles (AA), twelve heterozygous (AF) and six homozygous for weak alleles (FF) at the CSN1S1 locus, were used to evaluate the effect of genotype, diet and genotype × diet interaction on goat milk traits and casein profile. The goats were used in a 3 × 3 factorial arrangement of treatments, with three genotypes (AA, AF and FF) and three different energy intake levels: high (H), medium (M) and low (L). The diets supplied a complete pelleted feed containing 65% of alfalfa hay, respectively, at 150%, 100% and 70% of the total energy requirements. Milk yield was significantly affected by the genotype and diet: Lower levels were found in FF goats than in AA and AF genotypes (673.7 vs. 934.5 and 879.8 d/g, respectively; p = 0.002) as well as in goats fed with the L diet (651.5 vs. 1041 and 852.9 g/d for H and M diet, respectively, p < 0.001). The genotype influenced the casein profile. Specifically, AA goat milk exhibited higher concentrations of total casein and αs1-casein compared to AF and FF genotypes (for total casein and αs1-casein, respectively: 24.9 vs. 20.4 and 19.8 g/kg, p = 0.001; 7.2 vs. 3.7 and 0.7 g/kg, p < 0.001), while the FF genotype showed higher values for αs2-casein concentrations compared to homozygous AA and heterozygous AF goats (3.1 vs. 2.4 and 2.5 g/kg, respectively, p < 0.001). A significant genotype x diet interaction occurred for αs2-casein levels (g/kg) (p = 0.034) and αs1-casein yields (p = 0.027): The αs2-casein level was not affected by the diet in AA goats, whereas it increased with energy intake in AF and FF genotypes. Conversely, the αs1-casein yield gradually increased with energy intake in AA and AF groups, whereas the diet in FF goats did not modify it. The results demonstrated that high energy input, as well as the strong allele at the CSN1S1 locus, enhanced milk production and casein concentrations. Furthermore, they confirmed the existence of an interaction between αs1-casein polymorphism and diets, influencing the milk casein composition and yield.

Metagenomic, microbiological, chemical and sensory profiling of Caciocavallo Podolico Lucano cheese.

In this study, Caciocavallo Podolico Lucano (CPL) cheese was deeply characterized for its bacterial community, chemical composition and sensory aspects. The entire cheese making process (from milk collection to ripened cheese) was performed by strictly applying the traditional protocol for CPL production in four dairy factories (A-D) representative of the production area. The vat made of wood represents the main transformation tool for CPL cheese production and the biofilms hosted onto the internal surfaces of all vats analyzed in this study were dominated by lactic acid bacteria. Total mesophilic microorganisms present in bulk milk (4.7-5.0 log CFU/ml) increased consistently after contact with the wooden vat surfaces (5.4-6.4 log CFU/ml). The application of Illumina sequencing technology identified barely 18 taxonomic groups among processed samples; streptococci and lactobacilli constituted the major groups of the wooden vat biofilms [94.74-99.70 % of relative abundance (RA)], while lactobacilli dominated almost entirely (94.19-100 % of total RA) the bacterial community of ripened cheeses. Except coagulase positive staphylococci, undesirable bacteria were undetectable. Among chemical parameters, significant variations were registered for unsaturated, monounsaturated, polyunsaturated fatty acids and antioxidant properties (significantly lower for CPL cheeses produced in factory B). The cheeses from factories A, C and D were characterized by a higher lactic acid and persistence smell attributes than factory B. This work indicated that the strict application of CPL cheese making protocol harmonized the main microbiological, physicochemical and sensory parameters of the final cheeses produced in the four factories investigated.

Caciocavallo Podolico Cheese, a Traditional Agri-Food Product of the Region of Basilicata, Italy: Comparison of the Cheese's Nutritional, Health and Organoleptic Properties at 6 and 12 Months of Ripening, and Its Digital Communication.

Traditional agri-food products (TAPs) are closely linked to the peculiarities of the territory of origin and are strategic tools for preserving culture and traditions; nutritional and organoleptic peculiarities also differentiate these products on the market. One such product is Caciocavallo Podolico Lucano (CPL), a stretched curd cheese made exclusively from raw milk from Podolian cows, reared under extensive conditions. The objective of this study was to characterise CPL and evaluate the effects of ripening (6 vs. 12 months) on the quality and organoleptic properties, using the technological "artificial senses" platform, of CPL produced and sold in the region of Basilicata, Italy. Additionally, this study represents the first analysis of cheese-related digital communication and trends online. The study found no significant differences between 6-month- and 12-month-ripened cheese, except for a slight increase in cholesterol levels in the latter. CPL aged for 6 and 12 months is naturally lactose-free, rich in bioactive components, and high in vitamin A and antioxidants and has a low PUFA-n6/n3 ratio. The "artificial sensory profile" was able to discriminate the organoleptic fingerprints of 6-month- and 12-month-ripened cheese. The application of a socio-semiotic methodology enabled us to identify the best drivers to create effective communication for this product. The researchers recommend focusing on creating a certification mark linked to the territory for future protection.

Feeding Dairy Ewes with Fresh or Dehydrated Sulla (Sulla coronarium L.) Forage. 1. Effects on Feed Utilization, Milk Production, and Oxidative Status.

Feeding dairy ewes with fresh sulla forage (FSF), a legume species containing condensed tannins (CT), has been shown to increase feed intake, milk yield, and casein and enhances the oxidative status of animals. Dehydration of FSF could be an alternative to hay-making to preserve the nutritional properties. This research aimed to compare the responses of dairy ewes fed with diets based on sulla hay (SH), pelleted dehydrated sulla forage (DSF), or FSF in terms of efficiency of feed utilization, milk production, and the balance between oxidant (reactive oxygen metabolites (ROMs)) and antioxidant (biological antioxidant potential (BAP)) substances at the plasma level. Ten first-lambing (FL) and ten third-lambing (TL) ewes of the Valle del Belice breed at 60 days in milk were allocated into five homogeneous groups fed with different diets in a partial 5 × 2 Latin square design with two phases. The diets differed for the forage basis: SHL = SH ad libitum; DSF2 = 2 kg/day DSF per head plus SH ad libitum; FSF2 = 2 kg/day FSF per head plus SH ad libitum; FSF4 = 4 kg/day FSF per head plus SH ad libitum; FSFL = FSF ad libitum. A commercial concentrate was provided to FL (0.8 kg/day per head) and TL (1.2 kg/day per head) ewes. Dehydration induced slight variations in the content of protein and fiber, showed no loss of vitamin E and polyunsaturated fatty acids, and decreased the level of CT and polyphenols. The DSF2 diet resulted in a higher intake of dry matter, protein, and vitamin E compared to the other diets, whereas, compared to the FSFL diet, its intake was analogous for net energy and was lower in CT and polyphenols. The DSF2 diet was comparable to FSF4 and FSFL diets for milk yield, and to all diets for casein content and the clotting ability of milk. Ewes fed a DSF2 diet exhibited lowest values of ROMs and oxidative stress index (OSI = ROMs/BAP), indicating a better oxidative status, presumably due to the antioxidant protection exerted by the higher vitamin E intake and CT metabolites. These results confirmed the positive effects of FSF on milk production, especially due to CT intake, in improving the efficiency of dietary protein utilization, and showed how favorable effects also occur when FSF is replaced by DSF instead of SH.

The Quality of Five Natural, Historical Italian Cheeses Produced in Different Months: Gross Composition, Fat-Soluble Vitamins, Fatty Acids, Total Phenols, Antioxidant Capacity, and Health Index.

Five natural historic cheeses of Southern Italy were investigated-Caciocavallo Palermitano (CP), Casizolu del Montiferru (CdM), Vastedda della Valle del Belìce (VVB), Pecorino Siciliano (PS), and Caprino Nicastrese (CN)-which are produced with raw milk and with traditional techniques and tools, from autochthonous breeds reared under an extensive system. The effects of the month of production on gross composition, MUFA, PUFA, PUFA-ω6, PUFA-ω3, α-tocopherol, retinol, cholesterol, TPC, TEAC, and GHIC were evaluated. In CP, CLA, TPC, and GHIC were higher in April than in February. CdM showed higher values in terms of fat, saturated fatty acids, PUFA-ω3, α-tocopherol, TEAC, and GHIC in May than in February and September, while low values in terms of protein, moisture, and CLA were found. In VVB, MUFA, PUFA-ω6, and α-tocopherol increased in June compared with April; conversely, protein, FRAP, and TEAC were higher in April. In PS, protein, CLA, PUFA, PUFA-ω3, α-tocopherol, and GHIC increased in May compared with January; on the contrary, moisture, NaCl, and TEAC showed high values in January. CN showed higher values in terms of PUFA, PUFA-ω6, PUFA-ω3, TPC, TEAC, and GHIC in April and June compared with January. It is shown that each cheese is unique and closely linked to the production area. Cheeses produced in the spring months showed a high nutritional quality due to the greatest presence of healthy compounds originating from an extensive feeding system.

Local Small Ruminant Grazing in the Monti Foy Area (Italy): The Relationship Between Grassland Biodiversity Maintenance and Added-Value Dairy Products.

The literature indicates that grazing small ruminants, when adequately managed, contributes to grassland biodiversity maintenance. On the other hand, milk and cheese from grazing animals show higher nutritional and aromatic quality than those from stall-fed animals. The relationship between the two issues has rarely been addressed. This article provides information for a discussion of this relationship. First, two case studies are reported. Local breeds of small ruminants fed by grazing on pastures within the Special Area of Conservation "Monti Foy" in the Northwestern Basilicata region (Italy), with a stocking rate of 4.0 LU ha-1 year-1, showed the best effectiveness for the maintenance of grassland botanical biodiversity. Milk and cheese from pasture-fed goats showed higher contents of beneficial fatty acids, phenols, and vitamins A and E; higher degree of antioxidant protection; and richer volatile compound profiles, in particular for terpenes content. Finally, some recommendations for the management of grazing systems in similar mountain areas are offered, including a viable approach for land managers to preserve the grassland biodiversity of pastures and provide high-quality products that are valuable both for their nutritional quality and for their contribution to the economic sustainability of mountain communities.

Oxidative Status of Goats with Different CSN1S1 Genotypes Fed ad Libitum with Fresh and Dry Forages.

Forty late-lactation Girgentana goats were used to study the effect of diets fed ad libitum and αS1-casein (CSN1S1) genotype on redox balance. The goats genotyped at CSN1S1 locus (A/A, A/F) were subjected to four feeding treatments different for percentage inclusion of dry and fresh forage: DAF100 (98% of Dry Alfalfa Forage), DAF65 (65% of Dry Alfalfa Forage), FSF100 (100% of Fresh Sulla Forage) and FSF65 (65% of Fresh Sulla Forage). Blood samples were analyzed for superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity, reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP) and non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), albumin, glucose and cholesterol contents. The oxidative stress index (OSI) was calculated as percentage ratio of ROMs to BAP. Redox balance was improved by Sulla inclusion, as reflected in the lower OSI values found in FSF100 and FSF65 groups. DAF100 group displayed the highest GPX activity, while other groups exhibited the highest SOD activity. Fresh forage diets increased albumin concentration while no effect of tested factors was noted on glucose, NEFA, BHBA and cholesterol contents. The interaction diet × genotype was significant only for GPX activity. GPX and albumin were negatively correlated and were correlated positively and negatively with ROMs, respectively. Diet rather than genotype affects redox balance in dairy goats and a possible role of forage polyphenol compounds on oxidative status needs to be tested in future studies.

Improvement of Oxidative Status, Milk and Cheese Production, and Food Sustainability Indexes by Addition of Durum Wheat Bran to Dairy Cows' Diet.

Durum wheat bran (DWB) is a by-product mostly used in feeding ruminants, contributing to decrease in the utilization of feeds suitable as foods for human consumption, thus improving the sustainability of livestock production. However, the potential benefits of DWB, due to its content in phenolic acids, mainly consisting of ferulic acid with antioxidant properties, have not been well clarified yet. Accordingly, in this experiment, 36 lactating cows divided into three groups received, over a period of 100 days, one of three concentrates including DWB at 0% (DWB0), 10% (DWB10), or 20% (DWB20). The concentrates were formulated to be isoproteic and isoenergetic and, to balance the higher fiber content of the concentrates with DWB, the hay in the diets was slightly reduced. During the trial, the group feed intake and the individual milk production were monitored, and cheese was made with bulk milk from each group. Milk yield and microbiological characteristics of milk and cheese were similar among groups, indicating no DWB effect on cows performance and fermentation process. Milk from DWB20 group resulted slightly higher in casein and curd firmness (a2r). In cows fed DWB, the higher polyphenol intake was responsible for higher blood contents of these bioactive compounds, that seemed to have contributed in reducing the level of reactive oxygen metabolites (ROMs), which were higher in DWB0 cows. DWB20 cheeses showed a higher polyphenol content, lower number of peroxides, and higher antioxidant capacity than DWB0 cheeses. DWB20 and DWB10 diets resulted less expensive. In addition, the DWB20 group showed the best indexes heFCE (human edible feed conversion efficiency = milk/human edible feed) and NFP (net food production = milk - human edible food), expressed as crude protein or gross energy. In conclusion, the DWB fed to dairy cows at 12% of diet dry matter (DM) can lead to benefits, such as the improvement of oxidative status of cows, milk quality, shelf-life, and functional properties of cheese, and might contribute to reduce the feeding cost and limit the human-animal competition for feeding sources.

Leptin Gene Polymorphism in Goats Fed with Diet at Different Energy Level: Effects on Feed Intake, Milk Traits, Milk Fatty Acids Composition, and Metabolic State.

The study investigated the effects of a polymorphism at the LEP gene intron 1 microsatellite region and its interaction with diet energy level on feed intake, milk traits, milk fatty acid composition, and metabolic state in goats. Sixteen Girgentana lactating goats at mid-lactation, selected on the basis of their genotype (8 goats homozygous 266 bp/266 bp, L genotype; 8 goats heterozygous 266 bp/264 bp, H genotype), were fed ad libitum according to a change-over design, with two diets at different energy levels reached with different hay inclusion: low energy diet (LE)-100% of hay; and high energy diet (HE)-65% of hay. No differences in milk yield and composition or in dry matter intake were found between leptin genotypes or between diets. Leptin genotype had no effect on plasma metabolite concentrations. The differences between diets were recorded for plasma ß-hydroxybutyric acid (BHBA) concentrations with higher (p = 0.01) values for the HE compared to the LE diet (0.44 vs. 0.24 mmol/L, respectively). Nonesterified fatty acid (NEFA) values seem to indicate a positive energy balance in goats. No interaction genotype per diet was evident for most of the studied parameters. Fatty acid composition was strongly influenced by LEP genotype: L goats, compared to H goats, showed higher levels of monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and 14:1/14:0 desaturation index; lower levels of saturated fatty acids (SFA); and a more favorable atherogenic index. These results seem to suggest an improvement of health characteristics of milk with the L genotype.

Effects of Grazing on the Behaviour, Oxidative and Immune Status, and Production of Organic Dairy Cows.

This study compared the effects of a short daily grazing time with those of permanent free-stall housing on the behaviour, oxidative status, immune response, and milk production of organically reared cows. During a 63-day period, two homogeneous groups of eight lactating Brown cows were allocated to either housing (H) in a free-stall building for 24 h/day. Feeding was based on a total mixed ration or grazing (G) on barley grass for 5 h/day, and housing in a free-stall structure with feeding was based on the same total mixed ration offered to the H group. With regard to behaviour, H cows spent more time idling, walking, drinking, and self-grooming, whereas G cows showed a greater intent to eat and interact socially. Moreover, G cows exhibited slightly higher reactive oxygen metabolites and similar biological antioxidant potential concentrations than the H group, which indicates that short grazing resulted in an almost negligible increase in oxidative stress and an unchanged antioxidant capacity. Skin tests, performed by injecting phytohemoagglutinin intradermally, indicated that G cows had thicker skin than H cows at the end of the trial, an index of a better cell-mediated immune response. Grazing did not affect milk yield but improved milk quality in terms of an increase in fat and a reduction in urea content, somatic cell count, and total microbial count. Milk from G cows was richer in saturated fatty acids, likely because of the contribution of palmitic acid present in the grazed barley grass, and also showed higher contents of some healthy fatty acids, such as rumenic acid and α-linolenic acid, and a lower omega-6/omega-3 ratio. These results show that including a short grazing time in the diets of organic dairy cows does not have negative consequences for milk production and contributes to improved milk quality as well as to a more efficient immune response in the cows.

The Effect of Yerba Mate (Ilex Paraguariensis) Supplementation on Nutrient Degradability in Dairy Cows: An In sacco and In vitro Study.

This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.

Effect of CSN1S1 genotype and its interaction with diet energy level on milk production and quality in Girgentana goats fed ad libitum.

A study was carried out to evaluate how the energy level of the diet can affect milk production and quality in Girgentana lactating goats in relation to polymorphism at the alphas1-casein (CSN1S1) genotype locus. Twenty-seven goats, homogeneous for milk production (1.5+/-0.3 kg/d), days of lactation (90+/-10 d) and body weight (35.8+/-5.5 kg) were selected on the basis of their CSN1S1 genotype, as follows: nine goats homozygous for strong (AA) alleles, nine goats homozygous for weak alleles (FF) and nine goats heterozygous (AF). The goats were used in a 3x3 factorial arrangement of treatments, with three genotypes (AA, FF, AF) and three diets at different energy levels (100%, 65% and 30% of hay inclusion). The experiment consisted of three simultaneous 3x3 Latin squares for the three genotypes, with one square for each level of hay inclusion in the diet. All the animals were housed in individual pens. Each experimental period lasted 23 d and consisted of 15 d for adaptation and 8 d for data and sample collection, during which the goats received the scheduled diet ad libitum. The animals were fed three different diets designed to have the same crude protein content (about 15%) but different energy levels: a pelleted alfalfa hay (H100) and two feeds including 65% (H65) and 30% (H30) of alfalfa hay (respectively 1099, 1386 and 1590 kcal NE for lactation/kg DM). All the diets were ground and pelleted (6 mm diameter). AA goats were more productive than AF and FF goats (respectively: 1419 v. 1145 and 1014 g/d; P=0.002). Indeed the interaction energy levelxgenotype was significant (P=0.018): in fact AA goats showed their milk increase only when fed with concentrates. Differences in protein and in casein levels between the three genotypes were in line with results expected from the different allele contribution to alphas1-casein synthesis. Milk urea levels were significantly lower in AA goats compared with AF and FF genotypes (respectively 32.7 v. 40.4 and 40.4 mg/dl; P=0.049) and significantly lower when goats were fed with 65H and 30H diets than with 100H diet (respectively 37.4 and 34.3 v. 41.7 mg/dl; P<0.001). Indeed, a significant interaction genotypexdiet (P=0.043) occurred for milk urea, which was significantly lower in AA goats but only when fed with concentrates (65H and 30H). Blood concentrations of energy indicators (glucose, non-esterified fatty acids and beta-hydroxybutyric acid) were not influenced by genotype. The results confirm that strong alleles are associated with a greater efficiency of feed utilization and seem to show that a high energy level of the diet can further improve this efficiency.

Effects of plane of nutrition on oxidative stress in goats during the peripartum period.

This study aimed to monitor the effect of a high (HD; 140% of energy requirements) versus a low diet (LD; 80% energy requirements) on oxidative status in goats during the peripartum period. Blood samples were taken from all goats at -2, -1, 0 (partum), +2 and +4 weeks from delivery. Blood samples were assayed for their content of reactive oxygen metabolites (ROMs), thiol (SH) groups, total antioxidant capacity (OXY) and for glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities. The observation that ROMs levels significantly increased during the peripartum period was accompanied by a decrease of GSH-Px activity at weeks 2 and 4 postpartum, which suggested that the goats might have experienced some degree of oxidative stress and lipid peroxidation. Overall, changes to the nutritional level of the diet had very little or no effect on redox homeostasis. The lack of any correlation between the biomarkers measured indicated that each oxidative stress marker responded differently, indicating that redox homeostasis was impaired in these dairy goats during the peripartum period.