Effects of dietary arginine supplementation on muscle structure, meat characteristics and lipid oxidation products in lambs and its potential mechanisms of action.

This study aimed to assess the effect of dietary arginine supplementation on muscle structure and meat characteristics of lambs also considering lipid oxidation products and to contribute to reveal its mechanisms of action using tandem mass tagging (TMT) proteomics. Eighteen lambs were allocated to two dietary treatment groups: control diet or control diet with the addition of 1% L-arginine. The results revealed that dietary arginine supplementation increased muscle fibre diameter and cross-sectional area (P < 0.05), which was attributable to protein deposition, as evidenced by increased RNA content, RNA/DNA ratio, inhibition of apoptotic enzyme activity, and alterations in the IGF-1/Akt signaling pathway (P < 0.05). In addition, dietary arginine elevated pH24h, a* values, and IMF content, decreased shear force value and backfat thickness (P < 0.05), as well as decreased the formation of lipid oxidation products involved in meat flavor including hexanal, heptanal, octanal, nonanal and 1-octen-3-ol by increasing the antioxidant capacity of the muscle (P < 0.05). The proteomics results suggested that seven enrichment pathways may be potential mechanisms by which arginine affected the muscle structure and meat characteristics of lambs. In summary, arginine supplementation in lamb diets provides a safe and effective way to improve meat quality, and antioxidant capacity of muscle of lamb.

Effects of ultrasound on the structural and functional properties of sheep bone collagen.

The study evaluated the effect of an ultrasound-assisted treatment on the structural and functional properties of sheep bone collagen (SBC). The type and distribution of SBC were analyzed by proteome (shotgun) technology combined with liquid chromatography-tandem mass spectrometry. Compared with pepsin extraction, the ultrasound-assisted treatment significantly increased the collagen extraction rate by 17.4 pp (P < 0.05). The characteristic functional groups and structural integrity of collagen extracted by both methods were determined via Fourier transform infrared spectroscopy, ultraviolet absorption spectroscopy, and fluorescence spectroscopy. Circular dichroism spectra revealed that the ultrasound-assisted pretreatment reduced α-helix content by 1.6 pp, ß-sheet content by 21.9 pp, and random coils content by 28.4 pp, whereas it increased ß-turn content by 51.9 pp (P < 0.05), compared with pepsin extraction. Moreover, ultrasound-assisted treatment collagen had superior functional properties (e.g., solubility, water absorption, and oil absorption capacity) and foaming and emulsion properties, compared with pepsin extraction. Furthermore, the relative content of type I collagen in ultrasound-assisted extracted SBC was highest at 79.66%; only small proportions of type II, VI, X, and XI collagen were present. Peptide activity analysis showed that SBC had potential antioxidant activity, dipeptidyl peptidase 4 inhibitory activity, and angiotensin-converting enzyme inhibitory activity; it also had anticancer, antihypertensive, anti-inflammatory, and immunomodulatory effects.

Lipid transformation during postmortem chilled aging in Mongolian sheep using lipidomics.

This present study aimed to investigate the effects of postmortem chilled aging on lipid and molecular transformation mechanisms in Mongolian sheep. UPLC-ESI-MS/MS was used to identify 812 lipids in Mongolian sheep within four days of chilled aging. Using multivariate analysis, 95 significant differential lipids were identified. The levels of phosphatidylcholine, phosphatidylinositol, and certain free fatty acids were observed to rise significantly, while other phospholipids and acylcarnitines showed a downward trend. In addition, levels of most of the lysophospholipids increased significantly in the early postmortem period; however, their levels did not alter significantly after 48 h. The Kyoto encyclopedia of genes and genomes pathway analysis revealed that these differential lipids are primarily involved in glycerophospholipid metabolism, linoleic acid metabolism, arachidonic acid metabolism, and thermogenesis. Our results have important implications in terms of understanding lipid transformation and changes in meat quality during postmortem chilled aging in Mongolian sheep.

The Effect of the Wild Boar Hunting System on Agricultural Damages: The North-East of Italy as a Case Scenario.

Hunting is a method commonly used in several European countries to reduce crop damages by wild boar Sus scrofa. However, results are still controversial and poorly treated. Using data on official claims (i.e., damages to crops) and wild boar local counts and hunting bags collected from 2019 to 2022, the purpose of this work was to evaluate the effect of the hunting system (divided into eradication and non-eradication areas) carried out in the north-east of Italy on the number of agricultural damages. The spatio-temporal distribution of wild boar hunting, density and damages as well as the effect of hunting, were evaluated through the hotspot analysis and the zero-inflated models, respectively. The results obtained revealed no clear spatial overlap among wild boar hunting, wild boar density and damages to agriculture in both the eradication and non-eradication areas. Moreover, the current level of harvesting did not significantly affect the number of agricultural damages. A multifaceted approach focused on the numeric control of the species based on accurate local counts and oriented towards more age classes in a similar percentage should be preferred to mitigate damages to cultivars.

Transcriptome analysis reveals the molecular regulatory network of muscle development and meat quality in Sunit lamb supplemented with dietary probiotic.

Supplementing animal feed with probiotic additives can promote muscle production and improve meat quality. The study aimed to explore the effects of dietary probiotics supplementation on the performance, meat quality and muscle transcriptome profile in Sunit lamb. Overall, feeding probiotics significantly increased the body length, LT area, pH24h and intramuscular fat (IMF) content, but decreased cooking loss and meat shear force compared to the control group (P < .05). A total of 651 differentially expressed genes (DEGs) were found in probiotic supplemented lambs. Pathway analysis revealed that DEGs were involved in multiple pathways related to muscle development and fat deposition, such as the ECM-receptor interactions, the MAPK signaling pathway and the FoxO signaling pathway. Therefore, dietary probiotic supplementation can improve muscle development and final meat quality in Sunit lambs by altering gene expression profiles associated with key pathways, providing unique insights into the molecular mechanisms by which dietary probiotics regulate muscle development in the lamb industry.

Isolation, Purification and Structure Identification of a Calcium-Binding Peptide from Sheep Bone Protein Hydrolysate.

To isolate a novel peptide with calcium-binding capacity, sheep bone protein was hydrolyzed sequentially using a dual-enzyme system (alcalase treatment following neutrase treatment) and investigated for its characteristics, separation, purification, and structure. The sheep bone protein hydrolysate (SBPH) was enriched in key amino acids such as Gly, Arg, Pro, Leu, Lys, Glu, Val, and Asp. The fluorescence spectra, circular dichroism spectra, and Fourier-transform infrared spectroscopy results showed that adding calcium ions decreased the α-helix and ß-sheet content but significantly increased the random and ß-turn content (p < 0.05). Carboxyl oxygen and amino nitrogen atoms of SBPH may participate in peptide−calcium binding. Scanning electron microscopy and energy dispersive spectrometry results showed that SBPH had strong calcium-chelating ability and that the peptide−calcium complex (SBPH−Ca) combined with calcium to form a spherical cluster structure. SBPH was separated and purified gradually by ultrafiltration, gel filtration chromatography, and reversed-phase high-performance liquid chromatography. Liquid chromatography-electrospray ionization/mass spectrometry identified the amino acid sequences as GPSGLPGERG (925.46 Da) and GAPGKDGVRG (912.48 Da), with calcium-binding capacities of 89.76 ± 0.19% and 88.26 ± 0.25%, respectively. The results of this study provide a scientific basis for the preparation of a new type of calcium supplement and high-value utilization of sheep bone.

Effects of physical exercise on muscle metabolism and meat quality characteristics of Mongolian sheep.

The objective of this study was to investigate the effects of exercise training on muscle metabolism, fatty acid composition, carcass traits, and meat quality characteristics of Mongolian sheep. Fourteen Mongolian sheep were randomly divided into two groups (7 sheep in each) and placed in two adjacent livestock pens. One group of sheep was kept in the pen (Control [C] group) and the other group of sheep (Training [T] group) were driven away in a field to walk twice a day. The results showed a reduction in pH measured 45 min post mortem, L*, a*, and b* value, intramuscular fat, and carcass length, and an increase in the ultimate pH value and shear force in the meat of T group in comparison with that of C group (p < .050). Also, exercise training moderately affected the fatty acid composition of LT muscle. Compared with C group, the concentrations of myristoleic acid (C14:1) and stearic acid (C18:0) were increased (p < .050), while the concentrations of C20:3 n-6, neurolic acid (C24:1), and n-3 polyunsaturated fatty acid (PUFA) were decreased in T group (p < .050). Transcriptome analysis highlighted 621 genes differentially expressed in two groups, including 385 were up-regulated (e.g., GLUT4 and PGC-1α) and 236 were down-regulated (e.g., PLIN1 and ACSL3) in T with respect to C group. Besides, considering these genes, a number of enrichment pathways related to muscle metabolic processes, involving carbohydrate metabolism, lipid metabolism, oxidation reduction process, and muscle tissue development, were highlighted. In conclusion, these results contributed to a better understanding of the possible biological and molecular processes underlying the effects of exercise training on muscle metabolism and meat quality in Mongolian sheep, and provide useful information for contributing to understand the phenotypic and functional differences in meat quality of sheep.

Hempseed By-Product in Diets of Italian Simmental Cull Dairy Cows and Its Effects on Animal Performance and Meat Quality.

Cull dairy cows are important contributors to total beef production in the USA and in Europe. Hempseed cake is a by-product of oil production and it is rich in unsaturated fatty acids (FA). This study aimed to investigate the effect of adding hempseed cake to the diet of Italian Simmental (IS) cull dairy cows on performances and meat quality. Twenty-six cull dairy cows were divided into three dietary groups: hay-based, corn silage-based and pasture-based diets. Within each group, the animals were equally divided into two treatments according to the protein source of the concentrate: hempseed cake (HEMP) or soybeans meal (SB). The trial lasted four months. HEMP showed similar in vivo performance and carcass characteristics, such as average daily gain (p > 0.05) and dressing percentage (p > 0.05), compared with SB. Meat characteristics, such as ether extract content and Warner−Bratzler shear force, were also similar between experimental groups (p > 0.05). Considering FA composition, HEMP showed similar saturated FA and polyunsaturated FA content (p > 0.05) but lower desirable fatty acids (p < 0.05) content and a tendentially lower hypocholesterolemic/hypercholesterolemic ratio (p < 0.10) than SFA. Hempseed cake can substitute soybean in the diet of cull dairy cows without effects on performance or meat quality.

Gas Chromatography Combustion Isotope Ratio Mass Spectrometry to Detect Differences in Four Compartments of Simmental Cows Fed on C3 and C4 Diets.

Fatty acids (FAs) metabolism in animals represents an important field of study since they influence the quality and the properties of the meat. The aim of this study is to assess the possibility to discriminate the diets of cows in different animal compartments and to study the fate of dietary FAs in the bovine organism, using carbon isotopic ratios. Five FAs, both essential (linoleic and linolenic) and non-essential (palmitic, stearic, and oleic) in four compartments (feed, rumen, liver, meat) of animals fed two different diets (based on either C3 or C4 plants) were considered. For all compartments, the carbon isotopic ratio (δ13C) of all FAs (with few exceptions) resulted significantly lower in cows fed on C3 than C4 plants, figuring as a powerful tool to discriminate between different diets. Moreover, chemical reactions taking place in each animal compartment result in fraction processes affecting the δ13C values. The δ13CFAs tendentially increase from feed to meat in group C3. On the other hand, the δ13CFAs generally increase from rumen to liver in group C4, while δ13CFAs of rumen and meat are mostly not statistically different. Different trends in the δ13CFAs of the two groups suggested different FAs fates depending on the diet.

Animal Welfare and Farmers' Satisfaction in Small-Scale Dairy Farms in the Eastern Alps: A "One Welfare" Approach.

The multidimensional concept of animal welfare includes physical health, good emotional state, and appropriate behavior of the animals. The most recent methods for its assessment are inspired by the Welfare Quality, a project compiling animal-, resource-, and management-based measures. Recently, animal welfare assessment has also considered the human factor in a so-called "One Welfare" approach. The One Welfare framework highlights the interconnections between animal welfare, human well-being, and the environment. The concept seems to fit particularly well to mountain areas where the relationship between human, animals, and the environment is stronger. In such disadvantaged areas, farmers' well-being plays a key role in maintaining livestock farming profitably and sustainably. This study aims to investigate the relationship between farmers' satisfaction, animal welfare outcomes, and overall farm performance in 69 small-scale dairy farms in the Eastern Alps. Animal welfare assessment consisted of animal-based measures and was performed using the methodology proposed by the European Food Safety Authority for this type of farm. Moreover, the farmers were interviewed to retrieve data on farm characteristics and on their level of satisfaction toward workload, land organization, relationship with the agricultural/non-agricultural community, and the future of local agriculture. The results show that good animal welfare can be obtained in a mountain farming system. Clinical indicators show a low prevalence of diseases and of very lean cows as opposed to integument alterations. The workload is not perceived as a problem in traditional farms (i.e., tie-stall and with no participation in quality-certification schemes). Animal welfare is higher in those farms where farmers have a positive engagement with both the agricultural and non-agricultural community and where farmers are satisfied with their land organization. A One Welfare approach could be applied on a larger scale to fully understand the links between animal and human well-being in mountain areas.

The Return of Large Carnivores and Extensive Farming Systems: A Review of Stakeholders' Perception at an EU Level.

Conflicts between large carnivores and human activities undermine both the maintenance of livestock practices as well as the conservation of carnivores across Europe. Because large carnivore management is driven by a common EU policy, the purpose of this research was to assess stakeholders' perception towards bears and wolves at an EU level. We conducted a systematic search and subsequent analysis of 40 peer-reviewed studies collected from 1990 to September 2020 within Member States of the EU. Rural inhabitants and hunters exhibited the most negative attitude compared to urban inhabitants and conservationists, whose attitude was more positive. We showed that direct experience with predators as a consequence of ongoing re-colonization may have affected the degree of acceptance of certain categories and that the long-term coexistence between humans and carnivores does not necessarily imply increased tolerance. To encourage coexistence, we recommend monitoring changes in attitudes over time relative to carnivore population dynamics.

Fatty acids stable carbon isotope fractionation in the bovine organism. A compound-specific isotope analysis through gas chromatography combustion isotope ratio mass spectrometry.

The contribution of dietary fatty acids to the quality of the meat and their path through the bovine organism is currently the subject of a lot of research. Stable isotope ratio analysis represents a powerful tool for this aim, one that has not been studied in depth yet. In this work, for the first time, the carbon isotopic ratios of six fatty acids (myristic 14:0, palmitic 16:0, stearic 18:0, oleic 18:1n-9, linoleic 18:2n-6 and linolenic 18:3n-3 acids) in different matrixes (diet, rumen, duodenal content, liver and loin) were analysed through gas chromatography combustion isotope ratio mass spectrometry. Moreover, the quantification of the single fatty acids was carried out, providing important information supporting the carbon isotopic ratio results. The variation in the concentration of the fatty acids in the different matrices depends on the chemical modifications they undergo in the sequential steps of the metabolic path. GC-C-IRMS turned out to be a powerful tool to investigate the fate of dietary fatty acids, providing information about the processes they undergo inside the bovine organism.

Effect of Heat Stress on Dairy Cow Performance and on Expression of Protein Metabolism Genes in Mammary Cells.

The aim of this study was to assess the effect of heat stress on dairy cow performance and on the expression of selected genes involved in milk protein metabolism. Eight Italian Holstein Friesian cows were kept under thermoneutral conditions (temperature-humidity index (THI) < 72, CON) for 8 days and under mild heat stress conditions (72 < THI < 78, HS) for an additional 8 days. The rectal temperature, feed intake, and milk yield were recorded during the last 3 days of the CON and HS periods. During the same time period, milk samples were collected to assess the composition and expression of selected genes involved in milk protein metabolism. Gene expression analyses were performed on somatic cells from milk, which are representative of mammary tissue. In terms of dairy cow performance, HS resulted in lower milk and protein yields and feed intake but higher rectal temperature than for CON (p < 0.05). Under HS, there were greater abundances of HSPA1A (p < 0.05) and BCL2 (p < 0.05), compared to CON, but similar levels of CSN2 (p > 0.05), CSN3 (p > 0.05), HSPA8 (p > 0.05), and STAT5B (p > 0.05) mRNA. Mild heat stress reduced the performance of dairy cows without affecting the expression of genes coding for caseins.

Effect of Feeding Adaptation of Italian Simmental Cows before Summer Grazing on Animal Behavior and Milk Characteristics.

According to the alpine transhumance system, dairy cows are moved from indoor feeding with conserved forage to fresh herbage feeding on pasture. The aim of this study was to assess, as a feeding adaptation technique, the effect of a gradual inclusion of fresh herbage in the diet of Italian Simmental dairy cows before their transfer to alpine pasture on performance, behavior, and milk characteristics. Eighteen cows were assigned to three groups: animals transferred to alpine pasture with a 10-d feeding adaptation period consisting in gradual access to a pasture close to the valley farm (GT), animals transferred to alpine pasture without a feeding adaptation period (AT), and animals kept in the valley farm (IND). During the first two weeks of summer grazing, GT and AT showed higher rumination time and different concentrations of ketones, hydrocarbons, organic acids, toluene, alcohols, phenols, and dimethyl sulfone in milk as compared to IND, whereas no differences were found in milk yield, composition, or coagulation properties. No differences between GT and AT were evident for the studied variables. The feeding adaptation technique used in this study did not influence the performance and milk characteristics of Italian Simmental dairy cows grazing on alpine pasture.

Lactic Acid Bacteria: Variability Due to Different Pork Breeds, Breeding Systems and Fermented Sausage Production Technology.

Changes in the ecology of the various lactic acid bacteria (LAB) species, which are involved in traditional fermented sausages, were investigated in the light of the use of different breeds of pork, each of which was raised in two different environments and processed using two different technologies. The semi-quantitative molecular method was applied in order to understand how the different species alternate over time, as well as their concentration ratios. A significant increase in LAB over the first days of fermentation characterized the trials where the starter culture wasn't added (T), reaching values of 107-108 cfu g-1. On the other hand, in the trials in which sausages were produced with starter addition, LAB counts had a less significant incremental jump from about 106 cfu g-1 (concentration of the inoculum) to 108 cfu g-1. Lactobacillus sakei and Lb. curvatus were detected as the prevalent population in all the observed fermentations. Pediococcus pentosaceus, Lb. casei, Leuconostoc mesenteroides, Lactococcus garviae, and Lb. graminis also appeared, but their concentration ratios varied depending on the diverse experimental settings. The results of cluster analysis showed that a plant- and breed-specific LAB ecology exists. In addition, it was also observed that the breeding system can influence the presence of certain LAB species.

Environmental Sustainability Assessment of Dairy Farms Rearing the Italian Simmental Dual-Purpose Breed.

This study aimed to assess the environmental footprint of dairy farms rearing a dual-purpose breed, and to evaluate, through alternative scenario analyses, the fattening of calves and the cultivation of hemp as strategies for reducing the environmental impact of these farms. Eleven farms were evaluated for global warming (GWP), acidification (AC) and eutrophication (EUP) potential. The Life Cycle Assessment method with three scenarios, REAL, based on real data, BEEF, where calves were fattened in farm, and HEMP, where hemp was cultivated in farms, were considered. If referred to 1 m2 of utilizable agricultural land, the GWP, AC and EUP were 1.6 kgCO2eq, 21.7 gSO2eq and 7.1 gPO43-eq, respectively. If referring to 1 kg of fat and protein corrected milk, the emissions were 1.1-1.4 kgCO2eq, 14.8-19.0 gSO2eq, and 5.0-6.4 gPO43-eq, depending on the allocation method adopted. The emissions were associated positively with culling rate and negatively with production intensity. In BEEF and HEMP scenarios, the emissions were reduced by 8-11% and by 1-5%, respectively. Fattening the calves, evaluating the cultivation of alternative plants and improving the productive and reproductive efficiency of animals could be effective strategies for reducing the environmental footprint of the farm.

Steroids in pig hair and welfare evaluation systems: combined approaches to improve management in pig breeding?

The aim of the present pilot study was to determine the allostatic load by analysing the hair cortisol and dehydroepiandrosterone­sulphate (DHEA­S) of commercial pigs belonging to different farms having good overall animal welfare according to the CREnBA (Centro di Referenza Nazionale per il Benessere Animale - Brescia, Italy) assessment protocol. The study was conducted on 86 pigs belonging to three farms with a grade higher than 8 on the CReNBA welfare evaluation system. Hair samples were taken by shaving on sows 1­10 days after weaning (Farms 1 and 2) and at the age of 8­10 months (Farm 3). The hormone concentrations were measured by radioimmunoassay. From the box plots, it is evident that the hair cortisol concentrations of animals were different between farms. Conversely, the variability of the hair DHEA­S concentration was similar between the F1 and F2 farms but much lower at F3. For all the variables considered (cortisol, DHEA­S and cortisol/DHEA­S ratio), F2 showed a higher hair concentration level than F1 (P < 0.05). The study's results indicate that the measurement of cortisol and DHEA­S in pig hair shows a different allostatic load between them, although the official welfare evaluation method assessed the farms as having good overall animal welfare.

Dietary supplementation of Aspergillus oryzae meal and its effect on performance, carcass characteristics, blood variables, and immunity of broiler chickens.

This study investigated the effect of different levels and consumption periods of Aspergillus oryzae meal on performance, carcass characteristics, blood variables, and immunity of broiler chickens. A total of 270 (male and female) Ross 308 chicks were randomly assigned to 9 treatment groups. Two levels (2 g/kg diet and 4 g/kg diet as-fed) of Aspergillus oryzae meal (AO) and 4 consumption periods of AO (starter, grower, finisher, and entire period) in a 2 × 4 factorial arrangement were used. Compared with control, AO used during the entire rearing period increased weight gain, reduced relative weight of abdominal fat, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) serum levels, and increased antibody titers against influenza and Newcastle disease vaccination and sheep red blood cells injection. Few differences in the variables considered were found if AO was added to broiler diets only during specific consumption periods, and between the two supplementation levels of AO. In conclusion, the addition of AO to the broiler diet can have beneficial effects in terms of performance, carcass composition, and health, but these positive effects were mainly reached adding AO for the entire rearing period.

Cortisol, DHEA, and Sexual Steroid Concentrations in Fattening Pigs' Hair.

The aim of this study was to analyze the feasibility and reliability of using hair as a matrix to determine the dehydroepiandrosterone (DHEA) and sexual steroid concentrations and the cortisol/DHEA ratio in fattening pigs. The results could be also used to plan future research to identify threshold values in order to set up strategies to control the allostatic load and increase the resilience of fattening pigs before slaughter. The study was conducted on 107 commercial crossbred rearing pigs. The hair samples were taken by shaving at the age of 36 weeks, and concentrations of the hormones were measured using a solid-phase microtiter radioimmunoassay. Females had significantly higher cortisol levels (p < 0.01), significantly lower DHEA concentrations (p < 0.05) and significantly higher cortisol/DHEA ratios (p < 0.01) than barrows. Progesterone was significantly higher in gilts than in barrows (p < 0.01). Testosterone and 17ß-estradiol were significantly higher in barrows than in gilts (p < 0.05). If future research can produce threshold values for the different markers examined, the evaluation of animals under subclinical stress conditions will be possible.

Fatty Acid Profiles of Cow's Milk and Cheese as Affected by Mountain Pasture Type and Concentrate Supplementation.

The aim of this trial was to assess the effect of pasture type and concentrate supplementation on the fatty acids (FA) composition of milk and cheese obtained during summer grazing on mountain pasture. Seventy-two Italian Simmental dairy cows were assigned to two groups that differed by the amount of concentrate supplementation: 3.0 kg/head/d (HIGH) vs. 1.5 kg/head/d (LOW). The dairy cows grazed on a Poion alpinae alliance pasture (PAST1), and subsequently they grazed on a Seslerion caeruleae alliance pasture (PAST2) for 10 d each. In the last three days of each experimental period, milk samples were collected immediately before each cheese-making event. Cheese samples were collected from each cheese loaf after 60 d of ripening. LOW showed higher isoFA, FA intermediates of the ruminal biohydrogenation, C18:3 c9,c12,c15, and total polyunsaturated fatty acid (PUFA) levels than HIGH. The pasture type had a more limited effect on FA composition of milk than concentrate level and was mainly related to monounsaturated fatty acids (MUFA), which were higher in PAST1 than PAST2 (p < 0.05). In cheeses, these differences were reduced. The phytanic acid and phytanic isomer ratio (SRR/RRR) in milk were not affected either by supplement level (p > 0.05) or by type of pasture (p > 0.05). Increasing the concentrate offered to dairy cows from 1.5 to 3.0 kg/d did not markedly influence the level of PUFA in cheeses produced during summer grazing on high mountain pasture.