Role of hazelnut skin supplementation on plasma antioxidant status and cytokine profile in growing lambs.

In this study, the effect of hazelnut skin dietary supplementation on antioxidant status and cytokine profile was evaluated in growing lambs. A total of 22 male lambs at the age of 2 months, balanced for their initial live weight (15.33 ± SD 1.79 kg), were selected and allocated into two experimental groups: the control group (CON) receiving a maize-barley-based concentrated diet, and the hazelnut group (HS) receiving supplementation with hazelnut skin (150 g/kg on the dry matter) as a maize substitute for the concentrate diet. The experiment lasted for 56 days. Peripheral blood was collected at 7, 35, and 56 days of the experiment. The free radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the total antioxidant capacity assay (TAC), the reactive oxygen species (ROS), and reactive nitrogen species (RNS) were determined in plasma. The secretion of IL-1ß, IL-6, and IL-10 cytokines was also determined by ELISA. The DPPH was affected by the interaction between feeding strategy and time of sampling (p = 0.039) with a higher level of DPPH at 7 days in the HS group than the CON group. The time of sampling affected the levels of plasma TAC (p = 0.016), while the ROS/RNS levels showed a higher value in the HS group (p < 0.001), on average. The antioxidant/oxidant index, which combines the TAC and the ROS/RNS levels, was not affected by the inclusion of hazelnut skin in the diet (p = 0.394). The cytokine profile showed a lower IL-6 secretion at both 35 and 56 days than at 7 days, on average. Furthermore, the feeding treatment affected the IL-1ß level, showing a lower level in the HS group than in the CON group on average. Lambs from the HS group had higher IL-10 plasma levels than the CON group at 7 days of the experiment. The present data highlight an antioxidant effect and a modulatory role in the cytokine profile of HS supplementation in growing lambs.

First insight into extracellular vesicle-miRNA characterization in a sheep in vitro model of inflammation.

Extracellular vesicles (EVs) and their microRNA (miRNA) cargoes have garnered attention in the veterinary field for their regulatory role in various biological processes. This study aimed to (i) evaluate two techniques of EV isolation from sheep peripheral blood mononuclear cell (PBMC) supernatants using the ultracentrifugation (UC) and reagent (REA) methods and (ii) characterize the EV-miRNA profiles after an in vitro inflammatory environment mediated by lipopolysaccharides (LPS). Sheep peripheral blood was collected, and PBMCs were separated using a density gradient reagent. Subsequently, PBMCs were cultured at 37°C for 24 h (5% CO2), and the supernatants were collected to perform the EV isolation. The presence of CD81+ extracellular vesicle marker was determined, and the purity of isolated EVs was calculated as a ratio between the number of isolated EVs and the protein concentration. Moreover, the morphological characterization revealed mainly round-shaped structures with average sizes of 211 nm for EVs isolated by the UC method and 99 nm for EVs isolated by the REA method. Illumina NextSeq sequencing in a single-end mode was used to characterize the miRNA profile, and the differentially expressed (DE) miRNAs were analyzed using a combination of bioinformatics tools. The results revealed that the REA method is reliable for EV isolation from sheep supernatants. It was considered an improvement of the recovery rate and purity of EVs with the enhancement of the number and the expression levels of characterized miRNAs. The EVs isolated by the UC method after an LPS challenge showed 11 DE miRNAs, among which eight miRNAs were upregulated and three were downregulated. On the other hand, the REA method revealed an EV cargo in which eight DE miRNAs were upregulated and 21 DE miRNAs were downregulated. The master miRNA regulators of the biological process were identified by performing the MIRNA-mRNA network analysis, showing that, among the higher representative miRNAs based on the centrality and betweenness, the miR-26a-5p could have a crucial role in the resolution of inflammation. Moreover, the identification of the let-7 miRNA family in all the EVs showed potential targeted genes that regulate the inflammation and immune responses.

In-depth characterization of the sarcoplasmic muscle proteome changes in lambs fed with hazelnut skin by-products: Relationships with meat color.

This study investigated the effect of agro-industrial hazelnut skin by-products supplementation on lamb meat color variation and the changes in the sarcoplasmic muscle proteome during post-mortem storage (0, 4 and 7 days). Gel-based proteomics and bioinformatics approaches were applied to better understand the potential role of feeding strategies in modulating the mechanisms underpinning meat discoloration and post-mortem changes during storage. Therefore, twenty-two Valle del Belice male lambs were randomly assigned to two dietary treatments: control (C), lambs fed with maize-barley diet, and hazelnut skin (H), lambs fed hazelnut skin by-product as maize partial replacer in the concentrate diet. Hazelnut dietary treatment led to better lamb meat color stability as evidenced by the lowest decrease in redness and saturation index values. Proteomics and bioinformatics results revealed changes in the abundance of 41 proteoforms, which were mainly involved in glycolytic processes, responses to oxidative stress, and immune and endocrine system. The proteins allowed revealing interconnected pathways to be behind meat color variation as a consequence of using hazelnut skin by-products to sustainable feed lamb. The proteins can be used as potential predictors of lamb meat color variation. Accordingly, the regression equations developed in this paper revealed triosephosphate isomerase (TPI1) as a reliable candidate biomarker of color stability in lamb meat. SIGNIFICANCE: The use of agro-industrial by-products in animal feeding can be a potential sustainable strategy to reduce the environmental impacts of the food production chain and consequently improve animal welfare and product quality. The inclusion of hazelnut skin by-products in the animal's diet, due to the high concentration of polyphenols, represents an effective strategy to improve the oxidative stability of meat, with significant implications on color. The use of proteomics combined with bioinformatics on the sarcoplasmic proteome is a powerful approach to decipher the underlying mechanism. Accordingly, this approach allowed in this trial a deeper understanding of the molecular mechanisms involved in the post-mortem processes through the discovery of several biological pathways linked with lamb meat color variation. Glycolysis, followed by responses to oxidative stress, and other proteins involved in the immune and endocrine system were found as the major interconnected pathways that could act as potential predictors of lamb meat color stability. Candidate proteins biomarkers were further revealed in this study to be related with multiple meat color traits.

Effect of hazelnut skin by-product supplementation in lambs' diets: Implications on plasma and muscle proteomes and first insights on the underlying mechanisms.

This study aimed to evaluate the effect of hazelnut skin by-product supplementation on lamb meat quality characteristics and plasma and muscle proteomes. Twenty-two Valle del Belice male lambs were divided into two experimental groups: control (C), fed a maize-barley diet and hazelnut (H), fed hazelnut skin by-product as maize partial replacer in the concentrate diet. The meat of lambs fed hazelnut skin showed greater values of lightness, redness, yellowness, and chroma color parameters together with the highest myofibril fragmentation index. Two-dimensional electrophoresis and mass spectrometry applied on plasma proteome identified 20 protein spots corresponding to 18 unique gene names to be differently expressed due to hazelnut skin by-product substitution. For the early post-mortem muscle, 23 protein spots (42 unique gene names) were significantly up-regulated due to hazelnut skin by-product supplementation. Four proteins these being APOA1, PHB, ACTG1 and ALB, were found to be common to the two proteomes suggesting that these proteins could be candidate biomarkers to monitor in vita and post-mortem lamb meat quality traits. This study evidenced the main mechanisms involved in the supplementation of hazelnut skin by-product in lambs' diet and confirmed the possibility of using plasma proteome as a non-invasive way to predict lamb meat quality. SIGNIFICANCE: Maximizing the use of agro-industrial by-products as replacers of traditional feedstuff for improving animal products is one of the important challenges to preserving natural resources and guaranteeing environmental sustainability. Hazelnut (Corylus avellana L.) skin, obtained as a results of hazelnut roasting, represents a valuable by-products due to its high content in unsaturated fatty acids, tannins, and vitamins. Thus, including hazelnut skin by-product in small ruminant nutrition could reduce the costs of animal feedings for farmers as well as improve meat nutritional and sensorial characteristics. Additionally, monitoring the meat quality characteristics with fast, accurate, and non-invasive tools to find, before slaughter, animals with desired quality characteristics is of growing interest in the last years. In this regard, the objectives of this study were to assess i) the effect of hazelnut skin supplementation on lamb meat quality characteristics and plasma and muscle proteomes, and ii) whether analyzing plasma proteome by using a gel-based proteomic approach could effectively offer a more readily available option for determining lamb meat quality. Taken together, the proteomic approach applied to plasma and muscle proteomes, allowed us to reveal the pathways and the potential candidate plasma biomarkers to predict lamb meat production in the pre-slaughter phase.

First insights about the underlying mechanisms of Martina Franca donkey meat tenderization during aging: A proteomic approach.

This study aimed to evaluate the effect of different aging times (1, 6 and 14 days) on the tenderization rate and protein changes in ten Martina Franca donkey striploins using a proteomic approach. During aging, a progressive decrease in share force, hardness, gumminess, and chewiness together with an increase in myofibril fragmentation index were observed. Proteolysis monitored by immunoblotting revealed a progressive degradation of desmin and fast troponin-T over time and an increase of their degradation products up to 14 days aging. Proteomics revealed by means of two-dimensional electrophoresis 37 protein spots corresponding to 15 proteins to change significantly by increasing aging time. These proteins belong to three pathways, these being "muscle contraction, structure, and associated proteins" (9 proteins); ii) "energy metabolism" (5 proteins); and iii) "chaperone" (1 protein). This study is the first to highlight the possible role of interconnected pathways in driving the final quality of donkey meat and predicting its texture.

Cytokine Pattern of Peripheral Blood Mononuclear Cells Isolated from Children Affected by Generalized Epilepsy Treated with Different Protein Fractions of Meat Sources.

The objective of the present study was the evaluation of cytokine patterns in terms of TNF-α, IL-10, IL-6, and IL-1ß secretion in peripheral blood mononuclear cell (PBMC) supernatants isolated from blood of children affected by generalized epilepsy and treated in vitro with myofibrillar, sarcoplasmic, and total protein fractions of meat and fish sources. Children with generalized epilepsy (EC group, n = 16) and children without any clinical signs of disease, representing a control group (CC group n = 16), were recruited at the Complex Structure of Neuropsychiatry Childhood-Adolescence of Policlinico Riuniti (Foggia, Italy). Myofibrillar (MYO), sarcoplasmic (SA), and total (TOT) protein fractions were obtained from longissimus thoracis muscle of beef (BF) and lamb (LA); from pectoralis muscle of chicken (CH); and from dorsal white muscle of sole (Solea solea, SO), European hake (Merluccius merluccius, EH), and sea bass fish (Dicentrarchus labrax, SB), respectively. PBMCs were isolated from peripheral blood of EC and CC groups, and an in vitro stimulation in the presence of 100 µg/mL for each protein fraction from different meat sources was performed. Data were classified according to three different levels of cytokines produced from the EC group relative to the CC group. TNF-α, IL-10, and IL-6 levels were not affected by different meat fractions and meat sources; on the contrary, IL-1ß levels were found to be significantly affected by the tested proteins fractions, as well as different meat sources, in high-level cytokine group. On average, the protein fractions obtained from LB, BF, and CH meat sources showed a higher level of IL-1ß than the protein fractions obtained from EH and SB fish samples. When all cytokine classes were analyzed, on average, a significant effect was observed for IL-10, IL-1ß, and TNF-α. Data obtained in the present study evidence that the nutritional strategy based on protein from fish and meat sources may modulate the immunological cytokine pattern of infants with generalized epilepsy.

The future of functional food: Emerging technologies application on prebiotics, probiotics and postbiotics.

This review was the first to gather literature about the effect of emerging technologies on probiotic, prebiotic, and postbiotic products. Applying emerging technologies to probiotic products can increase probiotic survival and improve probiotic properties (cholesterol attachment, adhesion to Caco-2 cells, increase angiotensin-converting enzyme (ACE) inhibitory, antioxidant, and antimicrobial activities, and decrease systolic blood pressure). Furthermore, it can optimize the fermentation process, produce or maintain compounds of interest (bacteriocin, oligosaccharides, peptides, phenolic compounds, flavonoids), improve bioactivity (vitamin, aglycones, calcium), and sensory characteristics. Applying emerging technologies to prebiotic products did not result in prebiotic degradation. Still, it contributed to higher concentrations of bioactive compounds (citric and ascorbic acids, anthocyanin, polyphenols, flavonoids) and health properties (antioxidant activity and inhibition of ACE, α-amylase, and α-glucosidase). Emerging technologies may also be applied to obtain postbiotics with increased health effects. In this way, current studies suggest that emerging food processing technologies enhance the efficiency of probiotics and prebiotics in food. The information provided may help food industries to choose a more suitable technology to process their products and provide a basis for the most used process parameters. Furthermore, the current gaps are discussed. Emerging technologies may be used to process food products resulting in increased probiotic functionality, prebiotic stability, and higher concentrations of bioactive compounds. In addition, they can be used to obtain postbiotic products with improved health effects compared to the conventional heat treatment.

Postmortem Muscle Protein Changes as a Tool for Monitoring Sahraoui Dromedary Meat Quality Characteristics.

The effects of slaughter age (2 vs. 9 years) and postmortem time (6, 8, 10, 12, 24, 48 and 72 h) on the meat quality and protein changes of the longissimus lumborum muscles of the Algerian Sahraoui dromedary were investigated. Muscles of young dromedaries evidenced a slower acidification process and a significantly higher myofibrillar fragmentation index throughout the postmortem time. The SDS-PAGE of sarcoplasmic and myofibrillar proteins revealed that meat from young dromedaries was characterized by the lowest percentage of myoglobin (p < 0.001) and the highest percentage of desmin (p < 0.01). During postmortem time, a decrease was found for phosphoglucomutase (p < 0.01), α-actinin (p < 0.05) and desmin (p < 0.01) in meat from young dromedaries. Western blot revealed an intense degradation of troponin T in younger dromedaries, with an earlier appearance of the 28 kDa polypeptide highlighting differences in the proteolytic potential between dromedaries of different ages. Principal component analysis showed that meat from young dromedaries, starting from 24 h postmortem, was located in a zone of the plot characterized by higher levels of the myofibrillar fragmentation index, 30 kDa polypeptide and enolase, overall confirming greater proteolysis in younger animals. Data suggest that the investigation of the muscle proteome is necessary to set targeted interventions to improve the aging process of dromedary meat cuts.

Lamb Meat Quality and Carcass Evaluation of Five Autochthonous Sheep Breeds: Towards Biodiversity Protection.

In the present study, the evaluation of the carcasses and meat quality, in terms of chemical composition and fatty acid profile, of lambs from five autochthonous sheep breeds (Altamurana, Bagnolese, Gentile di Puglia, Laticauda, and Leccese) reared in continental Southern Italy, were studied. All the carcasses were evaluated according to the EU Mediterranean classification system for carcasses weighing less than 13 kg. Meat chemical composition and fatty acids profile were assessed on both loin and leg commercial cuts. Fatty acid composition of loin resulted in differences among breeds, displaying lower values of saturated fatty acid in Altamurana, Bagnolese, and Leccese breeds and the highest content of polyunsaturated fatty acid in the Altamurana breed. Principal component analysis grouped lamb according to fatty acid content and to conjugated linoleic acid (CLA), omega n-3 and n-6 fatty acids; thus, Altamurana, Bagnolese, and Leccese breeds are characterized by the highest values of CLA content. Our data demonstrated that lamb meat from autochthonous breed has good carcass quality and the content of CLA, n-3, and n-6 was valuable for human consumption; therefore, the valorisation of local meat quality can help to avoid the extinction of the autochthonous breed offering to the market and consumer's high nutritive products.

Synbiotic sheep milk ice cream reduces chemically induced mouse colon carcinogenesis.

Sheep dairy products containing prebiotic and probiotic ingredients may have health-promoting properties. Thus, this study evaluated the effects of sheep milk ice cream [conventional full-fat (CONV), full-fat enriched with probiotic (PROB, 100 mg % wt/wt of Lacticaseibacillus casei 01), or nonfat synbiotic (SYNB, Lacticaseibacillus casei 01 and inulin, 10% wt/wt)] on carcinogen-induced colonic crypt cytotoxicity and premalignant lesion development. Male Swiss mice received 2 doses of colon carcinogen azoxymethane (AOM, 15 mg/kg of body weight) at wk 3 and 4. Two weeks before and during AOM administrations (4 wk) mice were treated with CONV, PROB, or SYNB by gavage (10 mL/kg). Mice were euthanized at wk 4 or 25 (n = 5 or 10 mice/group, respectively). At wk 4, a significant reduction in micronucleated colonocytes was observed in PROB and SYNB groups, and a significant decrease in both p53 expression and apoptosis indexes in colonic crypts was observed in SYNB group. At wk 25, both PROB and SYNB interventions reduced the mean number of colonic premalignant lesions. However, only SYNB group showed lower incidence and number of high-grade premalignant lesions in the colonic mucosa. These findings indicate that PROB or SYNB sheep milk ice cream, especially SYNB intervention, can reduce chemically induced mouse colon carcinogenesis.

NETosis of Peripheral Neutrophils Isolated From Dairy Cows Fed Olive Pomace.

Neutrophils represent primary mobile phagocytes recruited to the site of infection, and their functions are essential to enhance animals' health performance. Neutrophils have an essential role in innate immunity and are able to kill the pathogens via the synthesis of neutrophil extracellular traps (NETs). The objective of the present work was the study of the in vitro NETosis of peripheral neutrophils isolated from dairy cows supplemented with olive pomace. Dairy cows (n = 16) balanced for parity (3.67 ± 1.5 for CON, 3.67 ± 1.9 for OP), milk yield (24.3 ± 4.5 kg d-1for CON and 24.9 ± 1.7 kg d-1 for OP), the number of days in milk (109 ± 83.5 for CON and 196 ± 51 for OP), and body weight (647 ± 44.3 kg for CON and 675 ± 70.7 kg for OP) were divided into two experimental groups fed with a control diet (CON) and supplemented with 6% of olive pomace (OP). Peripheral blood neutrophils were isolated and stimulated in vitro with phorbol-myristate-acetate (PMA) as a marker for activation and reactivity of the neutrophils. After isolation, both the viability and CD11b expression were analyzed by flow cytometry. Both NETosis by neutrophil elastase-DNA complex system and myeloperoxidase (MPO) activity were evaluated by ELISA. The specific antibodies against MPO and citrullination of Histone-H1 were used for investigating NETosis by immunofluorescence microscopy. The neutrophil elastase-DNA complexes produced during NETosis and MPO activity of neutrophil extracts were affected by OP supplementation. Furthermore, results from immunofluorescence analysis of NETosis depicted a similar result found by ELISA showing a higher expression of MPO and citrullination of Histone-H1 in OP than the CON neutrophils. In addition, all data showed that the OP diet resulted in a better response of neutrophils to PMA stimulation than the CON diet, which did not support the neutrophils' responses to PMA stimulation. Our results demonstrated that OP supplementation can enhance the neutrophil function in dairy cows leading to udder defense and inflammation response especially when an immunosuppression state can occur.

Nexus Between Immune Responses and Oxidative Stress: The Role of Dietary Hydrolyzed Lignin in ex vivo Bovine Peripheral Blood Mononuclear Cell Response.

The control of immune responses is particularly critical when an increase of oxidative stress occurs, causing an impairment of immune cell response and a condition of systemic inflammation, named oxinflammation. Nutritional strategies based on the use in the diet of phytochemicals extracted from plants, rich in antioxidants, could help restore the antioxidant/oxidant balance and obtain a modulation of immune response. Lignin represents a valuable resource of phenolic compounds, which are characterized by a corroborated antioxidant effect. To date, there are no studies reporting the effects of lignin in the diet on immune responses and oxidative stress in ruminants. The objective of the present experiment was the evaluation of the dietary inclusion of Pinus taeda hydrolyzed lignin on the ex vivo immune responses and oxidative stress biomarkers by peripheral blood mononuclear cells (PBMCs) isolated from beef steers. In order to test the effect during oxidative stress exposition, cells were treated with hydrogen peroxide (H2O2). The proliferation test and the viability assay were carried out on cells, whereas, on supernatants, the cytokine profile and the oxidative stress biomarkers were evaluated. The dietary inclusion with P. taeda hydrolyzed lignin resulted in cytoprotection after H2O2 exposition, increasing the number of viable monocytes and decreasing the reactive oxygen/nitrogen species production in supernatants. The cytokine profile indicated the modulatory role of hydrolyzed lignin on immune response, with a concomitant decrease of TNF-α and increase of IL-8 production, which are strictly connected with monocyte activation and antioxidant response pathway. These results demonstrated that hydrolyzed lignin may provide a modulation of oxidative stress and inflammatory response in PBMCs; thus, the P. taeda hydrolyzed lignin could be suggested as an innovative phytochemical in ruminant feed.

Effect of lipid fraction of digested milk from different sources in mature 3T3-L1 adipocyte.

We evaluated the effect of in vitro digested milk on mature adipocytes 3T3-L1, paying particular attention to its fatty acid composition, and comparing human (HM), donkey (DM), bovine (BM), ovine (OM), caprine (CM) and formula (FM) milk. Cellular viability, apoptosis, oxidative response and gene expression levels of NF-κB p65, HMGB1, SREBP-1c and FAS were evaluated. Digested milk treatments significantly reduced 3T3-L1 mature adipocytes viability and caspase activity compared with control group, but no significant differences were observed among different sources of digested milk. In all digested milk samples, ROS level was higher than the control, however, the digested human and formula milk showed lower levels of ROS than DM, BM, OM and CM samples. Lower capacity of HM and FM to induce oxidative stress in mature adipocytes was ascribed to the peculiar free fatty acids profile of digested milk samples. All milk treatments elicited a significant over-expression of NF-κB p65 in 3T3-L1 adipocytes compared to the control; the lowest gene expression was found in HM, BM, OM and CM, the highest in FM and an intermediate behavior was shown in DM. All digested milk treatments influenced the gene expression of SRBP-1c with FM and HM showing the highest levels. For FAS expression, BM showed the highest level, OM and CM intermediate and FM, HM and DM the lowest levels, however HM and DM had comparable levels to the control.

Ultrasound processing of fresh and frozen semi-skimmed sheep milk and its effects on microbiological and physical-chemical quality.

The objective of this study was to evaluate the effect of ultrasound treatment on the microbiological quality, protein and free amino acid profile of fresh and frozen stored semi-skimmed sheep milk. Milk was treated as fresh or frozen and stored up to one, three and six months. Output power time and pulse time were the parameters combined to design four different ultrasound (US) treatments: power 78 W and duration 6 min (US1); power 78 W and duration 8 min (US2); power 104 W and duration 4 min (US3) power 104 W and duration 6 min (US4). Pulse duration was of 4 s for each treatment. Sample US1 was discarded due to non effectiveness of US treatment, while other samples showed interesting results. Also, it was verified a frost effect on microorganisms in all samples which were frozen before treatment. No relevant change was reported on amino acid profile. The study showed promising results: the ultrasound treatment inactivated or eliminated the studied contaminant bacteria in semi-skimmed sheep milk, while maintained acceptable amount of lactic bacteria, which could be advantageous for dairy products processing.

Effect of Diet Supplementation with Quinoa Seed and/or Linseed on Immune Response, Productivity and Meat Quality in Merinos Derived Lambs.

In the last years several studies have investigated the strong relation between nutrition and immune response in the livestock production, particularly in dairy cattle and sheep. The aim of this study was to evaluate the effects of supplementation based on linseed, quinoa seeds and their combination on welfare, productivity and quality of meat from merinos derived lambs. 32 weaned lambs were divided into 4 experimental groups: quinoa (Q), linseed (LS) and combination of quinoa and linseed (LS + Q) that received the respective supplementation and control group (C) without supplementation. Lambs from all supplemented groups showed lower plasma urea, creatinine and cholesterol than control. Both linseed and quinoa supplementation enhanced the cell-mediated immune responses of lambs, furthermore, linseed supplementation resulted in the lowest level of cortisol secretion after handling, loading and transport. Meat from lambs supplemented with linseed and LS + Q showed the highest pH, at 1 and 3 h post-mortem, while, meat from all supplemented groups was more tender than meat from control. Results indicated that linseed and quinoa seeds supplementation can help the animal to cope with stressful events due to the close link between stress responses and the immune system and for improving meat quality in terms of better tenderness.

Focusing on fatty acid profile in milk from different species after in vitro digestion.

We report the fatty acid profile of raw milk and of the corresponding digested milk from different sources (human milk, formula milk and donkey, bovine, ovine and caprine milk) to gain information on the nutritional quality of different milk sources in infant nutrition.Short chain fatty acids (SC-FA) were higher in bovine and caprine milk, intermediate in ovine and donkey and lower in human and formula milk. Medium chain fatty acids (MC-FA) showed the highest values for bovine and caprine milk and the lowest for donkey and formula milk, whereas long chain fatty acids (LC-FA) were the highest in donkey and formula milk and intermediate in human milk.The percentage distribution of fatty acids liberated after in vitro digestion did not reflect the patterns found in the corresponding milk sources. In particular, MC free fatty acids (MC-FFA) showed the highest and the lowest values in donkey and in formula milk, LC-FFA showed the highest value in human milk. The total FFA was highest in human milk, lowest in formula milk and intermediate in donkey, bovine, ovine, and caprine milk.

Phytosterols from Dunaliella tertiolecta Reduce Cell Proliferation in Sheep Fed Flaxseed during Post Partum.

The post partum period is characterized by immunosuppression and increased disease susceptibility. Both phytosterols from microalga Dunaniella tertiolecta and dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) influence cell proliferation and cytokine release during inflammation. The objective of this paper was the evaluation of the effects of physterols, extracted and purified from D. tertiolecta, on the in vitro immune responses of ewes supplemented with flaxseed during post partum. Twenty Comisana parturient ewes were divided in two balanced groups, and supplemented with flaxseed (FS, 250 g/day) or fed with a conventional diet (CON). Blood samples (15 mL) were collected for five weeks, starting from lambing, in order to isolate peripheral blood mononuclear cells (PBMC). Stimulated PBMC were treated with a total sterols fraction from D. tertiolecta (TS), a mix of ergosterol and 7-dehydroporiferasterol (purified extract, PE), and a mix of acetylated ergosterol and 7-dehydroporiferasterol (acetylated purified extract, AcPE), extracted and purified from D. tertiolecta at two concentrations (0.4 and 0.8 mg/mL). Results of the experiment demonstrated that n-3 PUFA from flaxseed induced an anti-inflammatory cytokine profile, with an increase of both IL-10, IL-6 and a decrease of IL-1ß. TS, PE, and AcPE purified from D. tertiolecta showed an anti-proliferative effect on sheep PBMC regardless their chemical composition and concentration.

Bioactive Peptides in Animal Food Products.

Proteins of animal origin represent physiologically active components in the human diet; they exert a direct action or constitute a substrate for enzymatic hydrolysis upon food processing and consumption. Bioactive peptides may descend from the hydrolysis by digestive enzymes, enzymes endogenous to raw food materials, and enzymes from microorganisms added during food processing. Milk proteins have different polymorphisms for each dairy species that influence the amount and the biochemical characteristics (e.g., amino acid chain, phosphorylation, and glycosylation) of the protein. Milk from other species alternative to cow has been exploited for their role in children with cow milk allergy and in some infant pathologies, such as epilepsy, by monitoring the immune status. Different mechanisms concur for bioactive peptides generation from meat and meat products, and their functionality and application as functional ingredients have proven effects on consumer health. Animal food proteins are currently the main source of a range of biologically-active peptides which have gained special interest because they may also influence numerous physiological responses in the organism. The addition of probiotics to animal food products represent a strategy for the increase of molecules with health and functional properties.

Proteomic approach to investigate the impact of different dietary supplementation on lamb meat tenderness.

The aim of this study was to evaluate the effect of dietary supplementation of linseed and/or quinoa on tenderness and on proteome of lamb meat. Thirty-two Italian Merino lambs were distributed into 4 groups with different diet: control (CO) with no supplemental fat, linseed (LS), quinoa (QS) and QS+LS diets. Meat obtained by lamb fed linseed showed the lowest values of WBSF (P<0.001), hardness (P<0.01), gumminess (P<0.01) and chewiness (P<0.01). Proteomic changes of myofibrillar and sarcoplasmic proteins were estimated with SDS-PAGE, Western Blot and Two-Dimensional Gel Electrophoresis. In linseed group proteomic analysis revealed a degradation of desmin and TnT proteins complex and a major number of spots and phosphorylation isoforms of fast MLC2 patterns. Meat obtained by lamb fed quinoa showed a minor effect on the instrumental evaluation of meat tenderness and a major number of spots ascribed to sarcoplasmic proteins and fMHC. Data suggest that dietary supplementation may act on meat tenderness and on proteolytic pattern of myofibrillar fraction.

Lactoferrin Levels in Human Milk after Preterm and Term Delivery.

Background Lactoferrin (LF) is a highly represented, functional glycoprotein in human milk, exerting a wide range of anti-infective, immunomodulatory, and prebiotic actions in the neonate. Limited data are available assessing the concentrations and levels of LF in maternal milk over time during lactation in mothers who delivered infants at different GAs. Our aim with the present study was to determine the levels of LF in human milk from mothers of preterm and term infants and to evaluate the variations at a different time from delivery, in colostrum and mature milk. Methods Mothers of preterm and term infants from the Neonatology Unit in Foggia, Italy, were approached and enrolled in this study. From each mother, milk samples were collected within the first 3 days after birth (group A, 0-72 hours), between the 5th and 7th day after delivery (group B, 120-168 hours), and after the 10th day (group C, > 240 hours). All milk samples were divided into five groups, according to the GA of the infants: 24 to 27.6 weeks of GA (I), 28 to 31.6 weeks of GA (II), 32 to 34.6 weeks of GA (III), 35 to 37.6 weeks of GA (IV), and > 38 weeks of GA (V). Milk samples were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the content of LF. Results A total of 84 milk samples were collected from 28 mothers. We found that infant's GA, as well as the time of sampling, affected the levels of LF in milk. On one hand, LF showed higher content in human milk from group I (GA: 24-27.6 weeks) compared with the other groups (p < 0.01), and the levels of LF in colostrum were significantly correlated with GA (r = -0.31; p < 0.05). On the other hand, the LF content of milk had a significant decreasing trend over time. Overall, the highest values of LF were detected in preterm infants' maternal milk with a baby birth weight, lower than 1,400 g. Approximately 350 µg/mL was identified as the mean, physiological LF content in human mature milk in our population. Conclusions Levels of LF in human milk vary significantly over time during lactation and according to GA. This variability in the LF content of human milk may reflect the different needs of different infants during the early days and weeks of life. These data might help to inform models to design tailored supplementation strategies of LF in the nurseries and after home discharge.